bims-aporos Biomed news
on Apoptosis and reactive oxygen species
Issue of 2019‒03‒24
seventy-six papers selected by
Gavin McStay
Staffordshire University


  1. Chem Biol Interact. 2019 Mar 15. pii: S0009-2797(18)31701-0. [Epub ahead of print]
    Fang J, Tang Y, Cheng X, Wang L, Cai C, Zhang X, Liu S, Li P.
      BACKGROUND: Adriamycin (ADR) is an effective antineoplastic drug; the clinical application of ADR is limited due to fatal heart dysfunction. Exenatide has antioxidant, anti-apoptotic and anti-inflammatory properties. It can alleviate heart damage induced by ischaemia-reperfusion injury. Thus, we assumed that exenatide would produce protective effects on ADR-induced heart dysfunction.METHOD: Mice were treated with exenatide 1 h prior to every ADR treatment for 20 days. Left ventricular function and performance were assessed by echocardiography. Additionally, H9c2 cells were pretreated with exenatide followed by ADR, and intracellular reactive oxygen species (ROS) and cell viability, as well as the lactate dehydrogenase (LDH) and the creatine kinase MB (CK-MB), were subsequently measured. Flow cytometry and TUNEL staining were applied to assess the effect of exenatide on cardiac damage caused by ADR. Western blot and RT-PCR were performed to detect the effect of exenatide on apoptosis-related genes (Bcl-2 and Bax) and inflammation-related genes and/or proteins (tumour necrosis factor-α, interleukin-6, nuclear factor-κB, and p53).
    RESULT: Echocardiography showed that cardiac dysfunction caused by ADR was significantly improved by treatment with exenatide. ADR mice had harmful changes in the levels of ROS and CK-MB/LDH production, as well as the targeted apoptotic and inflammatory molecules, and these effects were also reversed by exenatide. In vitro, exenatide mitigated ADR-induced oxidative stress and CK-MB/LDH production, as well as Annexin V+/PI+ and TUNEL+ apoptosis in H9c2 cells.
    CONCLUSION: In conclusion, our research demonstrated the potential protective effects of exenatide on ADR-induced heart dysfunction through suppressing oxidative stress, apoptosis and inflammation.
    Keywords:  Adriamycin; Apoptosis; Exenatide; Heart dysfunction; Inflammation; Oxidative stress
    DOI:  https://doi.org/10.1016/j.cbi.2019.03.012
  2. Int J Biol Macromol. 2019 Mar 13. pii: S0141-8130(19)30467-2. [Epub ahead of print]
    Han Y, Nan S, Fan J, Chen Q, Zhang Y.
      Inonotus obliquus polysaccharide (IOPS) was initially separated and purified via precipitation from an aqueous extract with 80% alcohol, a DEAE-52 cellulose anion exchange column, and a Sephadex G-100 gel permeation chromatography system. IOPS was found to have a molecular weight of 111.9 kDa. In L-glutamic acid (L-Glu)-damaged HT22 cells, a 3-h pre-incubation with IOPS enhanced cell viability, inhibited apoptosis and caspase-3 activity, reduced the release of lactate dehydrogenase, restored the dissipated mitochondrial membrane potential, and suppressed the excess accumulation of intracellular reactive oxygen species. Compared with L-Glu-exposed cells, IOPS pre-treated cells exhibited reduced levels of Bcl-2 associated X protein (Bax) and Kelch-like ECH-associated protein 1 (Keap1) and enhanced levels of B-cell lymphoma-2 (Bcl-2), NF-E2p45-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), superoxide dismutase-1 (SOD-1), and cysteine ligase catalytic subunit. In amyloid precursor protein/presenilin 1 (APP/PS1) transgenic mice, an 8-week course of IOPS improved the pathological behaviors related to memory and cognition, reduced the deposition of β-amyloid peptides and neuronal fiber tangles induced by enhanced phosphor-Tau in the brain, and modulated the levels of anti- and pro-oxidative stress enzymes. Additionally, IOPS enhanced the expression levels of Nrf2 and its downstream proteins, including HO-1 and SOD-1, in the brains of APP/PS1 mice. The present study successfully demonstrated the protective effect of IOPS against AD and revealed the possible mechanism underlying the ability of IOPS to modulate oxidative stress, especially Nrf2 signaling, and mediate mitochondrial apoptosis.
    Keywords:  Alzheimer's disease; Apoptosis; Inonotus obliquus polysaccharides; Nrf2; Oxidative stress
    DOI:  https://doi.org/10.1016/j.ijbiomac.2019.03.033
  3. Med Sci Monit. 2019 Mar 22. 25 2132-2140
    Zhu Z, Li J, Zhang X.
      BACKGROUND Endothelial injury is the main mechanism of atherosclerosis, and is caused by oxidized low-density lipoprotein (ox-LDL). Astragaloside IV (AS-IV) is the primary active ingredient of the Chinese herb Huangqi, and exhibits antioxidant and anti-inflammatory properties in cardiovascular diseases. This study investigated the protective effect of AS-IV in human umbilical vein endothelial cells (HUVECs). MATERIAL AND METHODS HUVEC cells were induced with ox-LDL to establish an in vitro atherosclerosis model. Then HUVECs were pretreated for 1 h with AS-IV at different concentrations (10, 20, and 50 μM) and then exposed to ox-LDL (100 μg/mL) for 48 h. The cell viability, lactate dehydrogenase (LDH) release, apoptosis, migration, intracellular reactive oxygen species (ROS), and NADPH oxidase activity of HUVECs were measured. qRT-PCR was performed to measure the mRNA expressions of Nrf2, HO-1, TNFalpha, and IL-6. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the supernatant contents of TNFalpha and IL-6. RESULTS Exposure of HUVECs to ox-LDL reduced cell viability and migration, induced apoptosis, and increased intracellular ROS production and NADPH oxidase. Pretreatment with AS-IV (10, 20, and 50 μM) significantly enhanced the cell viability and migration, suppressed LDH release, apoptosis, ROS production, and NADPH oxidase in HUVECs, in a concentration-dependent manner. The AS-IV (50 μM) alone did not show significant differences from control. AS-IV increased mRNA expressions of Nrf2 and HO-1 and decreased mRNA expressions of TNFalpha and IL-6 in the ox-LDL-HUEVC cells. Furthermore, AS-IV reduced supernatant contents of TNFalpha and IL-6. CONCLUSIONS Astragaloside IV prevents ox-LDL-induced endothelial cell injury by reducing apoptosis, oxidative stress, and inflammatory response.
    DOI:  https://doi.org/10.12659/MSM.912894
  4. Int Immunopharmacol. 2019 Mar 19. pii: S1567-5769(19)30331-5. [Epub ahead of print]70 520-529
    Peng XP, Li XH, Li Y, Huang XT, Luo ZQ.
      Overactivation of the N-methyl-d-aspartate (NMDA) receptor promotes oxidative stress, aggravates the inflammatory response and induces excitotoxic lung injury. NMDA is a synthetic agonist that selectively activates the NMDA receptor. Oleanolic acid (OA) is a natural anti-inflammatory and antioxidant compound. This study investigated the effect and possible mechanism of OA on NMDA-induced acute lung injury (ALI) in mice. OA pretreatment alleviated NMDA-induced histological lung changes and ameliorated pulmonary oedema and pulmonary permeability. At the same time, OA inhibited inflammatory cell infiltration and decreased the levels of tumour necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1β in the lung and bronchoalveolar lavage fluid (BALF). OA markedly decreased malondialdehyde (MDA) production and increased the superoxide dismutase (SOD) and glutathione (GSH) contents of the lung in vivo. Meanwhile, we first found that NMDA increased LDH activity and decreased cell viability, and induced oxidative stress and apoptosis in mouse lung epithelial (MLE)-12 cells. By employing SRT1720 and sirtinol, the activator and inhibitor of sirtuin 1 (SIRT1), we found that SRT1720 partially eliminated the increase in ROS,and sirtinol further promoted the increase in ROS caused by NMDA. OA increased MLE-12 cells viability and attenuated oxidative stress after NMDA challenge in vitro. OA suppressed NMDA-induced MLE-12 cells apoptosis, while sirtinol inhibited the effect of OA. In addition, OA significantly upregulated the levels of SIRT1, nuclear-related factor 2(Nrf2) and Bcl-2 protein and downregulated the levels of acetylated nuclear factor-kappa B (NF-κB), NLRP3 and Bax protein. In conclusion, OA attenuated NMDA-induced excitotoxic lung injury, potentially through its anti-inflammatory, antioxidative stress and anti-apoptotic effects. The mechanism may be related to activating SIRT1 and reducing NF-κB acetylation.
    Keywords:  Apoptotic; Inflammation; N-methyl-d-aspartate; Nuclear factor-kappa B; Oleanolic acid; Reactive oxygen species; SIRT1
    DOI:  https://doi.org/10.1016/j.intimp.2019.03.018
  5. Eur J Pharmacol. 2019 Mar 14. pii: S0014-2999(19)30180-3. [Epub ahead of print]
    Liu Y, Han J, Zhou Z, Li D.
      Pancreatic β-cells are responsible for insulin secretion and control of plasma glucose levels. Accumulating evidences indicate a relationship between β-cell dysfunction/death and diabetes onset. Paeoniflorin (PF), a natural glycoside, has antihyperglycemic effect. However, the role of PF in pancreatic β-cells has not been examined. The aim of this study was to evaluate the protective effect of PF on streptozotocin (STZ)-induced β-cell damage. Our results showed that PF improved STZ-caused inhibitory effect on cell viability and insulin secretion ability in INS-1 cells. PF reduced caspase-3 activity and bax expression, and induced bcl-2 expression in STZ-treated INS-1 cells. PF resulted in a decrease in production of reactive oxygen species and MDA, and an increase in SOD activity in STZ-treated INS-1 cells. Furthermore, PF inhibited the phosphorylation of p38 and JNK, which is induced by STZ in INS-1 cells. The results suggested that PF protected INS-1 cells from STZ-induced cell damage. Meanwhile, PF suppressed the activation of p38 MAPK and JNK pathways in STZ-treated INS-1 cells. These results indicated that PF might be a natural anti-diabetic agent by improving pancreatic β-cells injury through inhibition of the p38 MAPK and JNK signaling pathways.
    Keywords:  Cell apoptosis; Diabetes mellitus (DM); MAPK signaling pathway; Pancreatic β-cells; oxidative stress; paeoniflorin (PF)
    DOI:  https://doi.org/10.1016/j.ejphar.2019.03.025
  6. J Environ Radioact. 2019 Mar 16. pii: S0265-931X(18)30563-0. [Epub ahead of print]203 112-116
    Soltani M, Zarei MH, Salimi A, Pourahmad J.
      Depleted uranium (DU) is a by-product of the enrichment procedure of natural uranium. During production and usage, uranium may be released into the environment due to failure to follow standard procedures, thus causing environmental pollution. In this study, toxicity effects of uranium (VI) and protective role of mitochondrial permeability transition pore sealing and antioxidant agents studied by isolated human lymphocytes. Human lymphocytes were exposed to different concentrations (0.1, 0.5, 1, 2 and 5 mM) of DU for 6 h and cytotoxicity was measured by trypan blue assay. The mechanistic parameters were assessed after 1, 2 and 3 h of lymphocyte treatment with 1/2 IC506h (0.3 mM), IC506h (0.8 mM) and 2 IC506h (1.6 mM) of DU. The reactive oxygen species (ROS), lysosomal membrane destabilization, mitochondrial membrane potential (MMP), lipid peroxidation, GSH and GSSG levels on human lymphocytes exposed to UA, were measured. The results indicate that toxicity of U (VI) was concentration dependent on human lymphocytes. Also, U (VI) induced ROS production, MMP reduction, lysosomal membrane destabilization and lipid peroxidation in human lymphocytes. In U (VI) treated lymphocytes, decrease in intracellular GSH and raise in extracellular GSSG levels were observed. We report that mitochondrial permeability transition (MPT) pore sealing and antioxidant agents, have the capacity significantly to prevents, mitochondrial toxicity. Thus, the inhibition of mitochondrial oxidative stress and mitochondrial dysfunction by MPT pore sealing and antioxidant agents is associated with the inhibition of DU-induced mitochondrial damages and activation of apoptosis in lymphocytes.
    Keywords:  (Uranium VI); Cytotoxicity; Depleted uranium; Human lymphocyte; Mitochondria
    DOI:  https://doi.org/10.1016/j.jenvrad.2019.03.009
  7. Nutrients. 2019 Mar 17. pii: E648. [Epub ahead of print]11(3):
    Muhammad T, Ikram M, Ullah R, Rehman SU, Kim MO.
      Glial activation and neuroinflammation play significant roles in apoptosis as well as in the development of cognitive and memory deficits. Neuroinflammation is also a critical feature in the pathogenesis of neurodegenerative disorders such as Alzheimer and Parkinson's diseases. Previously, hesperetin has been shown to be an effective antioxidant and anti-inflammatory agent. In the present study, in vivo and in vitro analyses were performed to evaluate the neuroprotective effects of hesperetin in lipopolysaccharide (LPS)-induced neuroinflammation, oxidative stress, neuronal apoptosis and memory impairments. Based on our findings, LPS treatment resulted in microglial activation and astrocytosis and elevated the expression of inflammatory mediators such as phosphorylated-Nuclear factor-κB (p-NF-κB), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in the cortical and hippocampal regions and in BV2 cells. However, hesperetin cotreatment markedly reduced the expression of inflammatory cytokines by ameliorating Toll-like receptor-4 (TLR4)-mediated ionized calcium-binding adapter molecule 1/glial fibrillary acidic protein (Iba-1/GFAP) expression. Similarly, hesperetin attenuated LPS-induced generation of reactive oxygen species/lipid per oxidation (ROS/LPO) and improved the antioxidant protein level such as nuclear factor erythroid 2-related factor 2 (Nrf2) and Haem-oxygenase (HO-1) in the mouse brain. Additionally, hesperetin ameliorated cytotoxicity and ROS/LPO induced by LPS in HT-22 cells. Moreover, hesperetin rescued LPS-induced neuronal apoptosis by reducing the expression of phosphorylated-c-Jun N-terminal kinases (p-JNK), B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax), and Caspase-3 protein and promoting the Bcl-2 protein level. Furthermore, hesperetin enhanced synaptic integrity, cognition, and memory processes by enhancing the phosphorylated-cAMP response element binding protein (p-CREB), postsynaptic density protein-95 (PSD-95), and Syntaxin. Overall, our preclinical study suggests that hesperetin conferred neuroprotection by regulating the TLR4/NF-κB signaling pathway against the detrimental effects of LPS.
    Keywords:  LPS; hesperetin; memory Impairments; microglia/astrocytes; neurodegeneration; neuroinflammation; reactive oxygen species (ROS); tumor necrosis factor (TNF)
    DOI:  https://doi.org/10.3390/nu11030648
  8. Free Radic Biol Med. 2019 Mar 14. pii: S0891-5849(19)30103-0. [Epub ahead of print]135 216-226
    Liu T, Zhang J, Han X, Xu J, Wu Y, Fang J.
      Cancer is considered as one of the highly mortal diseases globally. This is largely due to the lack of efficacious medicines for tumors, and thus development of potent anticancer agents is urgently needed. The thioredoxin (Trx) system is crucial to the survival ability of cells and its expression is up-regulated in many human tumors. Recently, increasing evidence has been established that mammalian thioredoxin reductase (TrxR), a selenocysteine-containing protein and the core component of the thioredoxin system, is a promising therapeutic target. The sesquiterpene lactone compound cynaropicrin (CYN), a major component of Cynara scolymus L., has shown multiple pharmacological functions, especially the anticancer effect, in many experimental models. Most of these functions are concomitant with the production of reactive oxygen species (ROS). Nevertheless, the target of this promising natural anticancer product in redox control has rarely been explored. In this study, we showed that CYN induces apoptosis of Hela cells. Mechanistic studies demonstrated that CYN impinges on the thioredoxin system via inhibition of TrxR, which leads to Trx oxidation and ROS accumulation in HeLa cells. Particularly, the cytotoxicity of CYN is enhanced through the genetic knockdown of TrxR, supporting the pharmacological effect of CYN is relevant to its inhibition of TrxR. Together, our studies reveal an unprecedented mechanism accounting for the anticancer effect of CYN and identify a promising therapeutic agent worthy of further development for cancer therapy.
    Keywords:  Apoptosis; Cynaropicrin; Oxidative stress; Reactive oxygen species (ROS); Redox regulation; Thioredoxin reductase
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.03.014
  9. Cells. 2019 Mar 20. pii: E263. [Epub ahead of print]8(3):
    Lin L, Liu Y, Fu S, Qu C, Li H, Ni J.
      Emodin is the main component of traditional Chinese medicines including rhubarb, Polygonum multiflorum, and Polygonum cuspidatum. It has confirmed hepatotoxicity and may be the main causative agent of liver damage associated with the above-mentioned traditional Chinese medicines. However, current research does not explain the mechanism of emodin in hepatotoxicity. In this study, L02 cells were used as a model to study the mechanism of emodin-induced hepatocyte apoptosis using quantitative proteomics, and the results were verified by Western blot. A total of 662 differentially expressed proteins were discovered and analyzed using Gene Ontology (GO) and pathway enrichment analysis. The results show that the oxidative phosphorylation pathway is highly represented. Abnormalities in this pathway result in impaired mitochondrial function and represent mitochondrial damage. This result is consistent with mitochondria membrane potential measurements. Analysis of differentially expressed proteins revealed that emodin mainly affects oxidative phosphorylation pathways by inhibiting the function of the mitochondrial respiratory chain complexes; the mitochondrial respiratory chain complex activity assay result also confirmed that emodin could inhibit the activity of all mitochondrial complexes. This results in an increase in caspase-3, a decrease in mitochondrial membrane potential (MMP,) an increase in reactive oxygen species (ROS), and disorders in ATP synthesis, etc., eventually leading to mitochondrial damage and hepatocyte apoptosis in vitro.
    Keywords:  complex; emodin; hepatotoxicity; mitochondrial; proteomic
    DOI:  https://doi.org/10.3390/cells8030263
  10. J Matern Fetal Neonatal Med. 2019 Mar 19. 1-141
    Zhu X, Lei X, Wang J, Dong W.
      BACKGROUND: Bronchopulmonary dysplasia (BPD) is one of the most common long-term lung complications of prematurely born infants caused by prolonged injury and repair during immature lung development. Resveratrol has reported to exert anti-inflammatory, antioxidation and antiapoptosis effects. However, it was not clear whether resveratrol played a protective role in rat model of BPD through antiapoptosis effect. This study aimed to investigate the effect of resveratrol in BPD.METHODS: Neonate rats were delivered spontaneously and randomized divided into four groups on postnatal day (PN) 0.5: room air (21% O2) + dimethyl sulfoxide (DMSO), room air + resveratrol, hyperoxia (80%) + DMSO, hyperoxia + resveratrol. Lung tissues were collected on PN1, PN7 and PN14. Protective effects of resveratrol on hyperoxia-induced lung injury were evaluated by hematoxylin and eosin (HE) staining, TUNEL staining, reactive oxygen species (ROS) detection, qRT-PCR and western blotting.
    RESULTS: Hyperoxia-induced alveolar simplification and apoptosis were alleviated by resveratrol; resveratrol reduced ROS production, up-regulated SIRT1, decreased the expressing of p53 and acetyl-p53 in the lung of hyperoxia-exposed neonatal rats.
    CONCLUSIONS: This study showed that resveratrol alleviated hyperoxia-induced apoptosis in neonatal rats lung tissue via reducing ROS and p53. Resveratrol induced SIRT1 upregulation and acetyl-p53 reduction may also be involved in lung protection.
    Keywords:  Apoptosis; SIRT1; bronchopulmonary dysplasia; p53; reactive oxygen species; resveratrol
    DOI:  https://doi.org/10.1080/14767058.2019.1597846
  11. J Investig Med. 2019 Mar 20. pii: jim-2018-000953. [Epub ahead of print]
    Huang L, Liu Z, Jiang H, Li L, Fu R.
      Most of the International Prognostic Scoring System (IPSS) high-risk patients with myelodysplastic syndrome partly responded to hypomethylating therapy even with transient remission, while arsenic trioxide (ATO) had partial effect in patients with MDS. Therefore, we sought to investigate the effects and possible mechanisms of the combination of ATO and decitabine (DAC) in MDS cells. In our study, the MUTZ-1 and SKM-1 cells were treated with ATO, DAC or both. Cell viability, cell apoptosis, levels of reactive oxygen species (ROS) and expressions of the endoplasmicreticulum (ER) stress-associated genes and proteins were examined. Results showed the combination of ATO and DAC synergistically inhibited the proliferation and induced apoptosis of MDS cells. Through the RNA-sequence and GSEA gene function analysis, ER stress-related pathway played an important role in apoptosis of MDS cells induced by the combination of ATO and DAC. ER stress-related genes DNA damage inducible transcript 3, GRP78, and activating transcription factor-6 were significantly highly expressed in combination group than those in single agent groups; proteins were confirmed by western blot. The levels of ROS significantly increased in the combination group. Furthermore, the apoptosis of (ATO+DAC) group MDS cells could be partially reversed by antioxidant agent N-acetylcysteine, accompanied by decreased expression of intracellular ROS and ER stress-related genes. These results suggested that the combination of ATO and DAC synergistically induced the apoptosis of MDS cells by increased ROS-related ER stress in MDS cells.
    Keywords:  apoptosis; arsenic trioxide; decitabine; endoplasmic reticulum stress; myelodysplastic syndrome
    DOI:  https://doi.org/10.1136/jim-2018-000953
  12. Brain Res. 2019 Mar 18. pii: S0006-8993(19)30159-3. [Epub ahead of print]
    Mohammadi AB, Torbati M, Farajdokht F, Sadigh-Eteghad S, Fazljou SMB, Mehdi Vatandoust S, Golzari SEJ, Mahmoudi J.
      This study was aimed to examine the effects of sericin administration on restraint stress induced anxiety- and depressive-like behaviors, oxidative stress, inflammation and apoptosis in the prefrontal cortex (PFC) and hippocampus (HIP) of mice. Animals were subjected to chronic restraint stress (3 h/day for 21 days) to induce a depressive-like model. Sericin was administered at different doses (100, 150, and 200 mg/kg/day, gavage for 21 days) along with immobilization. Elevated plus maze (EPM) and open field test (OFT) were performed to assess anxiety; while, the forced swim test (FST) and tail suspension test (TST) were implemented to evaluate depressive-like behaviors. Mitochondrial membrane potential (MMP), and markers of oxidative stress, neuroinflammation, and apoptosis were evaluated in the PFC and HIP regions. Moreover, serum levels of corticosterone were measured. Results showed that sericin increased number of central entries in OFT and prolonged time spent in open arms of EPM apparatus, while it reduced immobility time in TST and FST. Moreover, sericin treatments decreased oxygen species (ROS) and lipid peroxidation levels, restored MMP, and enhanced total antioxidant capacity (TAC) and enzyme activity of GPx and SOD in both brain regions. Furthermore, sericin reduced serum corticosterone concentration and suppressed neuroinflammatory response in the HIP and PFC, shown by decreased NF-κB, TNF-α, and IL-1β protein levels. Finally, sericin inhibited mitochondrial-dependent apoptosis pathway through down-regulation of Bax, cytochrome c, caspase-9 and -3, and up-regulation of Bcl-2 protein. These findings provide evidence for the protective effect of sericin therapy against psychopathological and behavioral changes induced by restraint stress.
    Keywords:  Apoptosis; Depression; Inflammation; Oxidative stress; Restraint stress; Sericin
    DOI:  https://doi.org/10.1016/j.brainres.2019.03.020
  13. Life Sci. 2019 Mar 16. pii: S0024-3205(19)30196-1. [Epub ahead of print]
    Li S, Sun Y, Han Z, Bu X, Yu W, Wang J.
      AIM: Atherosclerosis (AS) is a chronic condition of the arterial vessels and a risk factor for myocardial infarction and stroke. Euxanthone is a xanthone compound extracted from Polygala caudata, and shows vasodilatory action. The aim of this study was to determine the potential pharmacological effects of euxanthone against oxidized low-density lipoprotein (ox-LDL)-induced endothelial cell injury.MATERIAL AND METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to ox-LDL, following pre-treatment with different concentrations of euxanthone. Viability, apoptosis and DNA fragmentation were respectively assessed by CCK-8 assay, Annexin-V/PI staining and TdT-mediated dUTP Nick-End Labeling (TUNEL) assay. The cellular levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were analyzed by enzyme linked immune-sorbent assays (ELISA), and reactive oxygen species (ROS) levels using dichlorodihydrofluorescin diacetate (DCFH) staining. Quantitative RT-PCR and Western blotting were respectively used to analyze the expression levels of specific mRNAs and proteins. HUVECs were transfected with Nrf2 siRNA to induce knockdown of the latter.
    KEY FINDINGS: Euxanthone pre-treatment rescued the HUVECs from ox-LDL-induced cytotoxicity, apoptosis and DNA fragmentation in a dose-dependent manner. In addition, euxanthone also significantly reversed ox-LDL-triggered loss of mitochondrial membrane potential (MMP), cytochrome C release from mitochondria to cytosol, cleavage of caspase-3 and PARP, and increase in Bax/Bcl-2 ratio. Pre-treatment with euxanthone markedly suppressed ox-LDL-induced ROS generation and inhibition of antioxidant enzymes, as well as the up-regulation of pro-inflammatory factors like MCP-1, IL-1β and TNF-α in the HUVECs. Euxanthone up-regulated and activated Nrf2 by repressing Keap1, and increased the expression of its downstream genes HO-1 and NQO-1. Nrf2 knockdown abrogated the cyto-protective, anti-apoptotic, anti-oxidant and anti-inflammatory effects of euxanthone in ox-LDL-treated HUVECs. Finally, euxanthone activated Nrf2 via the MAPK pathway and blocking the latter likewise negated the protective effects of euxanthone against cell ox-LDL.
    SIGNIFICANCE: Euxanthone protected HUVECs against the oxidative and inflammatory damage induced by ox-LDL, indicating its potential as a novel therapeutic agent for AS.
    Keywords:  Atherosclerosis; ERK; Euxanthone; Nrf2
    DOI:  https://doi.org/10.1016/j.lfs.2019.03.032
  14. Evid Based Complement Alternat Med. 2019 ;2019 8798653
    Zhang W, Liu X, Jiang Y, Wang N, Li F, Xin H.
      Myocardial ischemia/reperfusion (I/R) injury is a key factor in deterioration of myocardial function. The c-Jun NH2-terminal kinase (JNK) activation and the transcription factor nuclear factor-kappaB (NF-κB) nuclear translocation have been found in I/R injury. 6-Gingerol, an important bioactive ingredient of ginger, has been reported to have cardiovascular pharmacological effects. However, the molecular mechanism through which it is beneficial is unclear. In this work, I/R induced the increase in the apoptosis and reactive oxygen species level in AC16 cardiomyocytes. 6-Gingerol administration decreased cardiomyocyte apoptosis and improved oxidative stress indexes. 6-Gingerol administration also inhibited I/R-induced HMGB2 expression upregulation and JNK activation and reduced Cleaved Poly(ADP-ribose) polymerases (PARP) and Caspase-3 expression. HMGB2 treatment mimicked the effect of I/R-induced cell damage, which was reversed by 6-gingerol administration. On the other hand, transcriptional activity of NF-κB was reduced in 6-gingerol treated cells. Thus, overall results indicated that 6-gingerol administration protected I/R-induced cardiomyocytes apoptosis via JNK/NF-κB pathway in the regulation of HMGB2. This work supported the efficacy of 6-gingerol on cardiovascular disease and partially revealed its mechanism, which was helpful for understanding the therapeutic effects of this natural drug.
    DOI:  https://doi.org/10.1155/2019/8798653
  15. Mol Med Rep. 2019 Mar 14.
    Fu X, Feng Y, Shao B, Zhang Y.
      Periodontal ligament stem cells (PDLSCs) are promising stem cells sources for regenerative medicine, particularly clinical periodontal ligament repair. It is critical to maintain high quality and a large quantity of PDLSCs for clinical usage. However, how PDLSCs respond to environmental stimuli, including reactive oxygen species (ROS), is poorly understood. The aim of the present study was to investigate how PDLSCs react to oxidative stress and the underlying mechanisms. Hydrogen peroxide‑induced oxidative stress was used to mimic a ROS increase in rat PDLSCs. The expression levels of Creb were detected under oxidative stress to examine the role that Creb serves in PDLSCs under oxidative stress. The present results demonstrated that the expression of Creb was reduced in a dose‑dependent manner in response to the H2O2 stimulus. Overexpressing Creb significantly reduced the ROS levels and protein expression levels of apoptotic genes in PDLSCs. The phosphorylation of the ERK pathway is indispensable in the activation of Creb‑induced protection. Our results revealed a protective role of Creb in ROS‑induced apoptosis, and validated the ERK/Creb/apoptosis regulator Bcl‑2 pathway works as an anti‑apoptotic signaling in PDLSCs. These findings will facilitate the in vitro culturing of PDLSCs for clinical usage and promote stem cell based therapy for periodontal tissue regeneration.
    DOI:  https://doi.org/10.3892/mmr.2019.10027
  16. Curr Eye Res. 2019 Mar 18.
    Tong L, Matsuura E, Takahashi M, Nagano T, Kawazu K.
      PURPOSE: We compare the cytotoxicity of anti-glaucoma prostaglandin ophthalmic solutions on human corneal epithelial cells, and elucidate mechanisms of toxicity.METHODS: Cell viability was examined using MTS assay, and morphological changes of the cells was observed. Induction of necrosis/apoptosis was measured by colorimetric caspase assay. The production of ROS and release of cytokines were analyzed using 2', 7'-dichlorodihydrofluorescein diacetate and bead-based indirect immunofluorescent assay, respectively.
    RESULTS: Xalatan, Lumigan 0.01% and Lumigan 0.03% decreased cell viability and induced morphological changes. Xalatan and Lumigan 0.01% induced necrosis. Xalatan, Lumigan 0.01%, Lumigan 0.03% and Taflotan stimulated ROS production. Travatan and Lumigan 0.03% increased concentrations of IL-6 and IL-8 in culture media.
    CONCLUSIONS: Xalatan and Lumigan 0.01% ophthalmic solutions demonstrated potent cytotoxicity compared with Lumigan 0.03%, Travatan, Taflotan and Taflotan UD. Taflotan UD, compared to Taflotan 0.0015%, induced less oxidative stress and apoptotic signalling. The cytotoxicity might be partly associated with benzalkonium chloride.
    Keywords:  Prostaglandin ophthalmic solution; cell death; human corneal epithelial cells; inflammatory cytokines; reactive oxygen species
    DOI:  https://doi.org/10.1080/02713683.2019.1597127
  17. Arch Toxicol. 2019 Mar 22.
    Li Q, Dong Z, Lian W, Cui J, Wang J, Shen H, Liu W, Yang J, Zhang X, Cui H.
      Ochratoxin A (OTA) is a common natural contaminant found in human and animal food worldwide. Our previous work has shown that OTA can cause oxidative DNA damage, G2 arrest and malignant transformation of human gastric epithelium (GES-1) cells. Mitochondria are considered to be target for the action of many cytotoxic agents. However, the role of mitochondria in the cytotoxicity of OTA remains unknown. The aim of this study is to explore the putative role of mitochondria on OTA cytotoxicity by analyzing mitochondrial changes in GES-1 cells. The results showed that OTA treatment (5, 10, 20 µM) for different times caused increases in the production of reactive oxygen species, and induced mitochondrial damage, shown by loss of mitochondrial membrane potential (ΔΨM), and decrease in cellular ATP concentration. Subsequently, the mitochondrial apoptotic pathway was activated, presented by increase of apoptotic rate and activation of apoptotic proteins. Autophagic cell death was also triggered, demonstrated by the conversion of light chain 3B (LC3B)-I to LC3B-II and elevated levels of green fluorescent protein-LC3 (GFP-LC3) puncta. Moreover, Parkin-dependent mitophagy was also activated presented by the colocalization of MitoTracker with LysoTracker or GFP-LC3 puncta. The inhibition of autophagy and mitophagy by inhibitors or siRNA attenuated the toxic effect of OTA on cell growth. Interestingly, OTA treatment also enhanced mitochondrial biogenesis confirmed by activation of AMPK/PGC-1α/TFAM pathway and promoted cell survival. Collectively, the effects of OTA on mitochondria of GES-1 cells are complex. OTA could cause mitochondrial function disturbance, apoptotic and autophagic cell death and also induce mitochondrial biogenesis.
    Keywords:  Apoptosis; Autophagy; Mitochondrial biogenesis; Mitochondrial dysfunction; Mitophagy; Ochratoxin A
    DOI:  https://doi.org/10.1007/s00204-019-02433-6
  18. J Mol Cell Cardiol. 2019 Mar 14. pii: S0022-2828(18)30810-1. [Epub ahead of print]
    Yang J, He J, Ismail M, Tweeten S, Zeng F, Gao L, Ballinger S, Young M, Prabhu SD, Rowe GC, Zhang J, Zhou L, Xie M.
      AIMS: The FDA-approved histone deacetylase (HDAC) inhibitor, suberoylanilide hydroxamic acid (SAHA, Vorinostat) has been shown to induce cardiomyocyte autophagy and blunt ischemia/reperfusion (I/R) injury when administered at the time of reperfusion. However, the precise mechanisms underlying the cardioprotective activity of SAHA are unknown. Mitochondrial dysfunction and oxidative damage are major contributors to myocardial apoptosis during I/R injury. We hypothesize that SAHA protects the myocardium by maintaining mitochondrial homeostasis and reducing reactive oxygen species (ROS) production during I/R injury.METHODS: Mouse and cultured cardiomyocyte (neonatal rat ventricular myocytes and human embryonic stem cell-derived cardiomyocytes) I/R models were used to investigate the effects of SAHA on mitochondria. ATG7 knockout mice, ATG7 knockdown by siRNA in cardiomyocytes and PGC-1α knockdown by adenovirus were used to test the dependency of autophagy and PGC-1α-mediated mitochondrial biogenesis respectively.
    RESULTS: Intact and total mitochondrial DNA (mtDNA) content and mitochondrial mass were significantly increased in cardiomyocytes by SAHA pretreatment before simulated I/R. In vivo, I/R induced >50% loss of mtDNA content in the border zones of mouse hearts, but SAHA pretreatment and reperfusion treatment alone reverted mtDNA content and mitochondrial mass to control levels. Moreover, pretreatment of cardiomyocytes with SAHA resulted in a 4-fold decrease in I/R-induced loss of mitochondrial membrane potential and a 25%-40% reduction in cytosolic ROS levels. However, loss-of-function of ATG7 in cardiomyocytes or mouse myocardium abolished the protective effects of SAHA on ROS levels, mitochondrial membrane potential, mtDNA levels, and mitochondrial mass. Lastly, PGC-1α gene expression was induced by SAHA in NRVMs and mouse heart subjected to I/R, and loss of PGC-1α abrogated SAHA's mitochondrial protective effects in cardiomyocytes.
    CONCLUSIONS: SAHA prevents I/R induced-mitochondrial dysfunction and loss, and reduces myocardial ROS production when given before or after the ischemia. The protective effects of SAHA on mitochondria are dependent on autophagy and PGC-1α-mediated mitochondrial biogenesis.
    Keywords:  Autophagy; HDAC inhibition; Mitochondrial homeostasis; Myocardial ischemia/reperfusion injury; ROS
    DOI:  https://doi.org/10.1016/j.yjmcc.2019.03.008
  19. Int J Oncol. 2019 Mar 19.
    Sun J, Feng Y, Wang Y, Ji Q, Cai G, Shi L, Wang Y, Huang Y, Zhang J, Li Q.
      α‑hederin, a monodesmosidic triterpenoid saponin, had previously demonstrated strong anticancer effects. In the current study, the pharmacological mechanism of autophagic cell death induced by α‑hederin was investigated in human colorectal cancer cells. First, through cell counting kit‑8 and colony formation assays, it was demonstrated that α‑hederin could inhibit the proliferation of HCT116 and HCT8 cell. Results of flow cytometry using fluorescein isothiocyanate Annexin V/propidium iodide and Hoechst 33258 staining indicated that α‑hederin could induce apoptosis. Western blotting demonstrated that α‑hederin could activate mitochondrial apoptosis signal pathway. Then, using light chain 3 lentiviral and electron microscope assay, it was demonstrated that α‑hederin could induce autophagy in colorectal cancer cells. In addition, immunohistochemistry results from in vivo experiments also demonstrated that α‑hederin could induce autophagy. AMP‑activated protein kinase (AMPK)/mechanistic target of rapamycin (mTOR) signaling was demonstrated to be activated by α‑hederin, which could be blocked by reactive oxygen species (ROS) inhibitor NAC. Furthermore, NAC could inhibit apoptosis and autophagy induced by α‑hederin. Finally, 3‑MA (autophagy inhibitor) reduced the inhibition of α‑hederin on cell activity, but it had no significant effect on apoptosis. In conclusion, α‑hederin triggered apoptosis through ROS‑activated mitochondrial signaling pathway and autophagic cell death through ROS dependent AMPK/mTOR signaling pathway activation in colorectal cancer cells.
    DOI:  https://doi.org/10.3892/ijo.2019.4757
  20. Biomed Res Int. 2019 ;2019 8727935
    Wang W, Niu S, Qiao L, Wei F, Yin J, Wang S, Ouyang Y, Chen D.
      Purpose: Multidrug resistance (MDR) is a major obstacle in chemotherapy of leukemia treatments. In this paper, we investigated Usnea Acid (UA) as MDR reversal agent on hematologic K562/ADR cells via ROS dependent apoptosis.Methods: CCK8 assay was used to measure cell viability rate of K562/ADR. Intracellular reactive oxygen species (ROS) generation, cell cycle distribution, cell apoptosis were measured with flow cytometry, respectively. Proteins related to apoptosis were measured by Western blot. Intracellular Adriamycin accumulation was observed by confocal microscopy and measured by flow cytometry.
    Results: In vitro study showed intracellular Adriamycin accumulation was remarkably increased by UA. Cell viability treated with Adr (4 μM) was decreased from 89.8%  ± 4.7 to 32%  ± 8.9 by combined with UA (4 μM). Adr-induced apoptosis and G1/G0 phase cell cycle arrest were remarkably increased by UA, as well as, intracellular ROS level. However, MDR reversing activity of UA was inhibited by N-acetyl cysteine (NAC), a ROS scavenger.
    Conclusion: These data provide compelling evidence that UA is a promising agent against MDR in leukemia cell line and suggest a promising therapeutic approach for leukemia.
    DOI:  https://doi.org/10.1155/2019/8727935
  21. Med Sci Monit. 2019 Mar 17. 25 2002-2008
    Zhang J, Gao Y, Ma C, Wang Y.
      BACKGROUND Murrayanine is a carbazole alkaloid derived from Murraya koenigii, which has been used in traditional Chinese medicine in the treatment of cancer. This study aimed to investigate the effects of murrayanine on human lung adenocarcinoma cells in vitro and to investigate the mechanisms of its action. MATERIAL AND METHODS A549 human lung adenocarcinoma cells and MRC-5 human lung fibroblasts were grown in culture, and an MTT assay determined cell viability. Cells were treated for 24 h with increasing doses of murrayanine (0, 9, 18, and 36 µM). Fluorescence, using 4', 6-diamidino-2-phenylindole (DAPI), acridine orange, ethidium bromide, and propidium iodide (PI), were used for the detection of apoptosis. The cell cycle was studied with fluorescence-activated cell sorting (FACS), and Western blot evaluated protein expression. RESULTS Murrayanine treatment resulted in significant dose-dependent inhibition of the growth of A549 cells (p<0.05), with an IC₅₀ of 9 µM, and arrested the cells at the G2/M phase of the cell cycle, reduced the expression of cyclin D and E, CDK2, 4, and 6, and increased the expression of p21 and p27. Murrayanine treatment increased apoptosis of the A549 cells and increased cleaved of caspase-3 and caspase-9, and the Bax/Bcl-2 ratio. Murrayanine treatment increased levels of reactive oxygen species (ROS), disrupted the mitochondrial membrane potential, inhibited invasion, and inhibited phosphorylation of p38 mitogen-activated protein kinase (MAPK) of the A549 cells. CONCLUSIONS Murrayanine induced cell cycle arrest, oxidative stress, and inhibited the expression of phosphorylated p38 in A549 adenocarcinoma cells.
    DOI:  https://doi.org/10.12659/MSM.913873
  22. J Inorg Biochem. 2019 Mar 14. pii: S0162-0134(19)30065-0. [Epub ahead of print]195 39-50
    Icsel C, Yilmaz VT, Cevatemre B, Aygun M, Ulukaya E.
      cis-[PtCl(sac)(PPh2Me)2] (1), cis-[PtCl(sac)(PPhMe2)2] (2), trans-[PtCl(sac)(PPh2Et)2] (3) and trans-[PtCl(sac)(PPhEt2)2] (4) complexes (sac = saccharinate) were synthesized and characterized by elemental analysis and spectroscopic methods. The structures of 2-4 were determined by X-ray single-crystal diffraction. The interaction of the complexes with DNA was studied various biochemical, biophysical and molecular docking methods. Only the cis-configured complexes (1 and 2) showed nuclease activity and their binding affinity towards DNA was considerably higher than those of their trans-congeners (3 and 4). The chlorido ligand in the cis-configured complexes underwent aquation, making them more reactive towards DNA. Furthermore, 1 and 2 exhibited anticancer potency on breast (MCF-7) and colon (HCT116) cancer cells similar to cisplatin, whereas 3 and 4 were biologicallly inactive. Mechanistic studies on MCF-7 cells showed that higher nuclear uptake, cell cycle arrest at the S phase, dramatically increased DNA double-strand breaks, apoptosis induction, elevated levels of reactive oxygen species (ROS) and high mitochondrial membrane depolarization greatly contribute to the anticancer potency of 1 and 2.
    Keywords:  Anticancer mechanism; Cytotoxicity; DNA binding; Phosphine; Pt(II) complex; Saccharinate
    DOI:  https://doi.org/10.1016/j.jinorgbio.2019.03.008
  23. Free Radic Biol Med. 2019 Mar 14. pii: S0891-5849(18)32634-0. [Epub ahead of print]135 235-244
    Hsu YH, Chuang HC, Lee YH, Lin YF, Chen YJ, Hsiao TC, Wu MY, Chiu HW.
      Traffic emission is responsible for most small-sized particulate matter (PM) air pollution in urban areas. Several recent studies have indicated that traffic-related PM may aggravate kidney disease. Furthermore, exposure to particulate air pollution may be related to the risk of chronic kidney disease (CKD). However, the underlying molecular mechanisms have not been adequately addressed. In the present study, we studied the mechanisms of renal damage that might be associated with exposure to PM. In a real world of whole-body exposure to traffic-related PM model for 3-6 months, PM in urban ambient air can affect kidney function and induce autophagy, endoplasmic reticulum (ER) stress and apoptosis in kidney tissues. Exposure to traffic-related diesel particulate matter (DPM) led to a reduction in cell viability in human kidney tubular epithelial cells HK-2. DPM increased mitochondrial reactive oxygen species (ROS) and decreased the mitochondrial membrane potential. Furthermore, DPM induced ER stress and activated the unfolded protein response (UPR) pathway. Eventually, DPM exposure induced caspase pathways and triggered apoptosis. In addition, DPM induced autophagy through the inhibition of the Akt/mTOR pathway. Autophagy inhibition resulted in significantly increased cytotoxicity and apoptosis. These findings suggest that air pollution in urban areas may cause nephrotoxicity and autophagy as a protective role in PM-induced cytotoxicity.
    Keywords:  Apoptosis; Autophagy; Endoplasmic reticulum stress; Nephrotoxicity
    DOI:  https://doi.org/10.1016/j.freeradbiomed.2019.03.008
  24. Int J Mol Med. 2019 Mar 19.
    Lee HJ, Spandidos DA, Tsatsakis A, Margina D, Izotov BN, Yang SH.
      The aim of this study was to investigate the antioxidant and anti‑apoptotic activities, as well as the underlying mechanisms of action, of Scrophularia buergeriana (S. buergeriana) extract (SBE) in glutamate‑induced SH‑SY5Y cell death. The roots of S. buergeriana were extracted with 70% ethanol, and standardized SBE was used in this study. To induce cytotoxicity, the SH‑SY5Y cells were exposed to glutamate for 3 h, or pre‑treated with SBE for 1 h, and subsequently incubated with glutamate for 3 h. The neuroprotective effects were assessed by measuring cell viability and the total glutathione contents using commercial kits. The antioxidant and anti‑apoptotic mechanisms of action of SBE were evaluated by western blot analysis. The results confirmed that glutamate‑induced toxicity was caused by reactive oxygen species (ROS) production, leading to oxidative stress and DNA damage, thus leading to cell death. However, treatment of the SH‑SY5Y cells with SBE significantly increased the viability of the cells exposed to glutamate by upregulating the levels of antioxidant proteins, such as superoxide dismutase (SOD)1, SOD2 and glutathione peroxidase‑1 (GPx‑1), and directly enhancing the total glutathione contents. Furthermore, SBE attenuated DNA impairment and decreased B‑cell lymphoma-2 (Bcl‑2)‑associated X protein (Bax), cleaved caspase‑3 and cleaved poly(adenosine diphosphate (ADP)‑ribose) polymerase (PARP) activation. In addition, SBE upregulated Bcl‑2 expression via p38 mitogen‑activated protein kinases (MAPKs). On the whole, the findings of this study demonstrated that SBE exerts neuroprotective effects against glutamate‑induced cell toxicity through its antioxidant and anti‑apoptotic activities.
    DOI:  https://doi.org/10.3892/ijmm.2019.4139
  25. J Cell Biochem. 2019 Mar 19.
    Zhu R, Wang W, Yang S.
      Cryptotanshinone (CTS), an active component extracted from the root of Salvia miltiorrhiza Bunge , was reported to attenuate hepatic ischemia/reperfusion (I/R) injury. However, its protective effect against renal I/R injury remains unclear. In this study, the role of CTS in renal I/R injury in vitro and its possible mechanism were investigated. Our results showed that CTS improved cell viability in HK-2 cells exposed to hypoxia/reoxygenation (H/R). CTS also inhibited the H/R-mediated production of reactive oxygen species, as well as increased the activities of superoxide dismutase and catalase in H/R-stimulated HK-2 cells. In addition, CTS dramatically attenuated the induction of bax expression and caspase-3 activity and alleviated the reduction of bcl-2 expression in HK-2 cells cultured with H/R. Furthermore, CTS activated the levels of p-PI3K and p-Akt in H/R-injured HK-2 cells; meanwhile, the renal protective activity of CTS was inhibited by the inhibitor of the (phosphatidylinositol 3 kinase/protein kinase B) PI3K/Akt pathway (LY294002). These findings indicate that CTS can ameliorate renal I/R injury in vitro partly through regulating the PI3K/Akt pathway.
    Keywords:  acute kidney injury; cryptotanshinone; hypoxia/reoxygenation; ischemia/reperfusion injury; oxidative stress
    DOI:  https://doi.org/10.1002/jcb.28609
  26. Med Mol Morphol. 2019 Mar 16.
    Tamaru S, Kajihara T, Mizuno Y, Takano N, Tochigi H, Sato T, Ishihara O.
      Clinical trials have shown that administering heparin during the luteal phase has beneficial effects on implantation and live birth rates. Heparin exerts direct effects on decidual human endometrial stromal cells (HESCs), which are independent of its anticoagulant effect. However, the accurate effects of heparin on the decidualization process remain unidentified. Here, we demonstrate that HESCs become dramatically resistant to oxidative stress upon decidualization, and we hypothesize a possible direct action of heparin on the decidualization of HESCs, which would lead to improved implantation. To test this hypothesis, we established primary HESC cultures and propagated them, and then we decidualized confluent cultures with 8-bromo-cAMP, with medroxyprogesterone acetate, and with or without heparin. We treated the cells with hydrogen peroxide (H2O2) as a source of reactive oxygen species (ROS). Adding heparin to decidualized HESCs induced prolactin secretion. Decidualized HESCs treated with heparin were prevented from undergoing apoptosis induced by oxidative stress. Heparin induced nuclear accumulation of the forkhead transcription factor FOXO1 and expression of its downstream target, the ROS scavenger superoxide dismutase 2. These results demonstrate that heparin-treated decidualized HESCs acquired further resistance to oxidative stress, suggesting that heparin may improve the implantation environment.
    Keywords:  Endometrial decidualization; FOXO1; Heparin; Oxidative stress; SOD2
    DOI:  https://doi.org/10.1007/s00795-019-00220-x
  27. Front Neurosci. 2019 ;13 26
    Teng L, Fan L, Peng Y, He X, Chen H, Duan H, Yang F, Lin D, Lin Z, Li H, Shao B.
      Carnosic acid (CA) has been reported to exhibit a variety of bioactivities including antioxidation, neuroprotection, and anti-inflammation; however, the impact of CA on subarachnoid hemorrhage (SAH) has never been elucidated. The current study was undertaken to explore the role of CA in early brain injury (EBI) secondary to SAH and the underlying mechanisms. Adult male Sprague-Dawley rats were perforated to mimic a clinical aneurysm with SAH. CA or vehicle was administered intravenously immediately after the SAH occurred. Mortality, SAH grade, neurologic function scores, brain water content, Evans blue extravasation, and the levels of reactive oxygen species (ROS) levels in the ipsilateral cortex were determined 24 h after the SAH occurred. Western blot, immunofluorescence, Fluoro-Jade C (FJC) and TUNEL staining were also performed. Our results showed that CA decreased ROS levels, alleviated brain edema and blood-brain barrier permeability, reduced neuronal cell death, and promoted neurologic function improvement. To probe into the potential mechanisms. We showed that CA increased SIRT1, MnSOD, and Bcl-2 expression, as well as decreased p66shc, Bax, and cleaved caspase-3 expression. Interestingly, sirtinol, a selective inhibitor of SIRT1, abolished the anti-apoptotic effects of CA. Taken together, these data revealed that CA has a neuroprotective role in EBI secondary to SAH. The potential mechanism may involve suppression of neuronal apoptosis through the SIRT1/p66shc signaling pathway. CA may provide a promising therapeutic regimen for management of SAH.
    Keywords:  apoptosis; carnosic acid; early brain injury; p66shc; subarachnoid hemorrhage
    DOI:  https://doi.org/10.3389/fnins.2019.00026
  28. Phytother Res. 2019 Mar 19.
    Hwang-Bo H, Lee WS, Nagappan A, Kim HJ, Panchanathan R, Park C, Chang SH, Kim ND, Leem SH, Chang YC, Kwon TK, Cheong JH, Kim GS, Jung JM, Shin SC, Hong SC, Choi YH.
      Evidence suggests that auranofin (AF) exhibits anticancer activity by inhibiting thioredoxin reductase (TrxR). Here, in this study, we have investigated the synergistic effects of AF and morin and their mechanism for the anticancer effects focusing on apoptosis in Hep3B human hepatocellular carcinoma cells. We assessed the anticancer activities by annexin V/PI double staining, caspase, and TrxR activity assay. Morin enhances the inhibitory effects on TrxR activity of AF as well as reducing cell viability. Annexin V/PI double staining revealed that morin/AF cotreatment induced apoptotic cell death. Morin enhances AF-induced mitochondrial membrane potential (ΔΨm) loss and cytochrome c release. Further, morin/AF cotreatment upregulated death receptor DR4/DR5, modulated Bcl-2 family members (upregulation of Bax and downregulation of Bcl-2), and activated caspase-3, -8, and -9. Morin also enhances AF-induced reactive oxygen species (ROS) generation. The anticancer effects results from caspase-dependent apoptosis, which was triggered via extrinsic pathway by upregulating TRAIL receptors (DR4/DR5) and enhanced via intrinsic pathway by modulating Bcl-2 and inhibitor of apoptosis protein family members. These are related to ROS generation. In conclusion, this study provides evidence that morin can enhance the anticancer activity of AF in Hep3B human hepatocellular carcinoma cells, indicating that its combination could be an alternative treatment strategy for the hepatocellular carcinoma.
    Keywords:  Hep3B cells; ROS; apoptosis; auranofin; cancer; morin
    DOI:  https://doi.org/10.1002/ptr.6329
  29. Nutr Neurosci. 2019 Mar 22. 1-11
    Tseng YT, Hsu HT, Lee TY, Chang WH, Lo YC.
      OBJECTIVES: Recent studies revealed the neuroprotective effects of naringenin (NGEN), a common dietary bioflavonoid contained in citrus fruits. However, there are limited data on its protection against methylglyoxal (MG), the most potent precursor of advanced glycation end-products. The present study was to investigate the protection of NGEN on MG-induced neurotoxicity and the involvement of insulin-like growth factor 1 receptor (IGF-1R) signaling.METHODS: NSC34 motor neuron-like cells was used. Cell viability was measured by MTT assay. Protein expressions were analyzed by western blots. Morphological changes of neurites were observed by an inverted microscope. Reactive oxygen species (ROS) production and apoptotic cell numbers were measured by flow cytometer. Glutathione (GSH) level and superoxide dismutase (SOD) activity were measured by ELISA.
    RESULTS: NGEN attenuated ROS production and increased GSH level, SOD activity and nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear expression in MG-treated NSC34 cells. NGEN also increased neurite length and enhanced IGF-1R and p-Akt in MG-treated NSC34 cells. Furthermore, NGEN attenuated MG-induced apoptotic death accompanied with down-regulation of cleaved-poly (ADP-ribose) polymerase (PARP) and up-regulation of B-cell lymphoma-2 (Bcl-2). However, AG1024, an IGF-1R antagonist, attenuated the anti-oxidative and anti-apoptotic effects of NGEN in MG-treated cells.
    DISCUSSION: The present results demonstrated that NGEN decreased neuronal apoptosis and improved antioxidant defense in MG-treated NSC34 cells. Moreover, IGF-1R-mediated antioxidant defense plays an important role in this protective mechanism. These findings suggest the potential benefits of NGEN on the prevention of MG-induced or diabetes/hyperglycemia-related neurotoxicity. In vivo studies are needed for further confirmation on NGEN-mediated neuroprotection.
    Keywords:  Naringenin; anti-apoptosis; antioxidant defense; glucose neurotoxicity; insulin-like growth factor 1; methylglyoxal; motor neurons
    DOI:  https://doi.org/10.1080/1028415X.2019.1594554
  30. Mol Biol (Mosk). 2019 Jan-Feb;53(1):53(1): 101-108
    Petrushanko IY, Melnikova EV, Yurinskaya MM, Vinokurov MG, Suslikov AV, Mitkevich VA, Makarov AA.
      Lipopolysaccharides (LPS), components of the cell wall of gram-negative bacteria, activate neutrophils that trigger pathological processes, including gram-negative sepsis. LPS inhibit spontaneous apoptosis of neutrophils that leads to inflammation. In this work we tested the action of H2S donor (GYY4137) on the activation of human neutrophils by E. coli LPS. We estimated the changes in redox status (ROS level, intracellularglutathione, NO), apoptosis and mitochondrial potential of neutrophils under the LPS action in the presence and absence of GYY4137. GYY4137 reduces the ROS level, slightly reduces GSH, does not influence the NO level and has no apoptogenic effect. LPS induce the increasing of ROS level and inhibit spontaneous apoptosis of neutrophils. We found that GYY4137 prevents the growth of ROS caused by LPS and leads to a reduction of LPS-induced inhibition of neutrophil apoptosis. Thus the mechanism of GYY4137 protection against inflammation, triggered by bacterial infection, is concerned with the neutralization of LPS effect on neutrophils.
    Keywords:  H2S donor GYY4137; apoptosis; glutathione; lipopolysaccharides; neutrophils; reactive oxygen species
    DOI:  https://doi.org/10.1134/S0026898419010130
  31. Mol Neurobiol. 2019 Mar 22.
    He JN, Zhang SD, Qu Y, Wang HL, Tham CC, Pang CP, Chu WK.
      Glaucoma is a chronic optic neuropathy that could lead to permanent vision loss. Primary open-angle glaucoma (POAG) is the most common type of glaucoma, with elevated intraocular pressure (IOP) as a major risk factor. IOP is mainly regulated by trabecular meshwork (TM), an important component of the conventional aqueous humor (AH) outflow pathway. TM cells are constantly subjected to oxidative stress. Long-term exposure to oxidative stress has been shown to cause elevation of AH outflow resistance, leading to higher IOP. In this study, we induced chronic oxidative stress in human trabecular meshwork (TM-1) cells with 1 μM rotenone and investigated the levels of reactive oxygen species (ROS), autophagy, and mitochondrial functions. Protective effects of rapamycin, an inducer of autophagy, were also investigated. Our data indicated that rotenone significantly increased oxidative stress, but not autophagy, in TM-1 cells. Rapamycin at 10 nM effectively suppressed the rotenone-induced cell apoptosis, as well as the ROS elevation. The protective effects of rapamycin could be associated to the induction of autophagy and removal of damaged mitochondria in TM-1 cells. Our results suggest autophagy has important roles in protecting TM-1 cells from oxidative stress, which could be further developed into a novel treatment to POAG.
    Keywords:  Autophagy; Oxidative stress; POAG; Trabecular meshwork
    DOI:  https://doi.org/10.1007/s12035-019-1559-5
  32. Saudi J Biol Sci. 2019 Mar;26(3): 531-540
    Al-Basher GI.
      Iron overload toxicity was shown to associate with chronic liver diseases which lead to hepatic fibrosis and subsequently the progression to cancer through oxidative stress and apoptotic pathways. Green tea potential activity as chelating, anti-oxidative, or anti-apoptotic mechanisms against metal toxicity was poorly clarified. Here, we are trying to evaluate the anti-oxidant and anti-apoptotic properties of green tea in the regulation of serum hepcidin levels, reduction in iron overloads, and improve of liver fibrosis in iron overloaded experimental rats. Three groups of male adult rats were randomly classified into three groups and treated as follows: control rats, iron treated rats for two months in drinking water followed by either vehicle or green tea extract (AGTE; 100 mg/kg) treatment for 2 more months. Thereafter, we studied the effects of AGTE on iron overload-induced lipid peroxidation, anti-oxidant depletion, liver cell injury and apoptosis. Treatment of iron-overloaded rats with AGTE resulted in marked decreases in iron accumulation within liver, depletion in serum ferritin, and hepcidin levels. Iron-overloaded rats had significant increase in malonyldialdehyde (MDA), a marker of lipid peroxidation and nitric oxide (NO) in liver when compared to control group. Also, significant change in cytochrome c and DNA content as apoptotic markers were reported in iron treated rats. The effects of iron overload on lipid peroxidation, NO levels, cytochrome c and DNA content were significantly reduced by the intervention treatment with AGTE (P < 0.001). Furthermore, the endogenous anti-oxidant capacities/levels (TAC) in liver were also significantly decreased in chronic iron overload and administration of AGTE restored the decrease in the hepatic antioxidant activities/levels. Also, hepatic hepcidin was shown to be significantly correlated with oxidative and apoptotic relating biomarkers as well as an improvement in liver fibrosis of iron treated rats following AGTE treatment. In-vitro analysis showed that, the improvement in iron toxicity of the liver depend mainly on antioxidant and protective ability of green tea polyphenolic compounds especiallyepigallocatechin-3-gallate (EGCG). Our study showed that green tea extract (GTE) ameliorates iron overload induced hepatotoxicity, apoptosis and oxidative stress in rat liver via inhibition of hepatic iron accumulation; improve of liver antioxidant capacity, and down regulation of serum hepcidin as well as reduction in the release of apoptotic relating proteins.
    Keywords:  Apoptosis; Green tea; Hepcidin; Iron overload; Liver fibrosis; Oxidative stress
    DOI:  https://doi.org/10.1016/j.sjbs.2017.08.007
  33. Front Pharmacol. 2019 ;10 175
    Zhang H, Liu M, Zhang Y, Li X.
      Exhausted exercise has been reported to cause the damage of myocardial structure and function in terms of cardiomyocyte apoptosis, oxidative stress, and energy metabolism disturbance. Trimetazidine (TMZ), as an anti-ischemic agent, has been approved to be effective in promoting myocardial energy metabolism, anti-inflammatory, and anti-oxidation. However, few studies examined the effects of TMZ on myocardial injury induced by exhausted exercise. To investigate whether TMZ could ameliorate the exhaustive exercise-induced myocardial injury and explore the underlying mechanisms, here the rat model of exhaustive exercise was induced by prolonged swimming exercise and TMZ was administrated to rats before exhaustive exercise. According to the results, we demonstrated that exhaustive exercise led to cardiomyocyte damage in rats as evidenced by elevations in cTnI and NT-proBNP levels, and decrease in CX43 expression, which was attenuated by TMZ treatment. Moreover, the administration of TMZ was found to restrain exhaustive exercise-induced oxidative stress damage by increasing GSH level, SOD and GSH-Px activities, and decreasing MDA level. Additionally, TMZ ameliorated myocardial injury by inhibiting apoptosis via reducing Bax/Bcl-2 ratio and down-regulating cleaved caspase-3, cleaved PARP, and cytochrome c levels in the myocardium of rats. Furthermore, we found that TMZ suppressed oxidative stress and cardiomyocyte apoptosis via activation of Nrf2/HO-1 and inactivation of NF-κB signaling pathways. Therefore, our study suggested that TMZ provided cardioprotection in rats after exhaustive exercise, indicating TMZ might served as a potential therapeutic drug for exhaustive exercise-induced myocardial injury.
    Keywords:  apoptosis; cardioprotection; exhaustive exercise; myocardial injury; oxidative stress; trimetazidine
    DOI:  https://doi.org/10.3389/fphar.2019.00175
  34. Biotechnol Appl Biochem. 2019 Mar 20.
    Zhu L, Li D, Chen C, Wang G, Shi J, Zhang F.
      Nerve damage is the main pathogenesis of neurodegenerative diseases. Recently, in search for a promising therapeutic target that could stop Neurodegenerative diseases progression, the anti-oxidant signaling pathway regulated by transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) has attracted new hopes. Icariin (ICA) exhibited a battery of pharmacological properties, including anti-oxidation, anti-aging, anti-inflammation activities. Recent studies indicate ICA conferred neuroprotection against brain ischemic injury and neurodegenerative diseases. However, the mechanisms underlying ICA-mediated neuroprotection remain unelucidated. This study aimed at analyzing whether ICA evoked neuroprotection against 6-hydroxydopamine (6-OHDA)-induced neurotoxicity in PC12 cells and the mechanisms of action. ICA protected against 6-OHDA-induced neuronal damage, accompanied by the inhibition of cell apoptosis through the marked decreases in the Bax/Bcl-2 ratio, cytochrome C release and caspase-3 cleavage. In addition, the activation of Nrf2 signaling pathway was responsible for ICA-mediated neuroprotection. First, ICA relieved reactive oxygen species (ROS) accumulation and increased superoxide dismutase (SOD) generation via the activation of Nrf2 signaling. Second, Nrf2 knockdown by siRNA reversed ICA-mediated neuroprotection. These experiments offer a promising avenue to validate Nrf2 for a compelling target with ICA as a therapeutic strategy to enhance endogenous brain defense mechanisms against Neurodegenerative diseases. (Delete this sentence and change to "Together, these results suggested ICA-mediated neuroprotection might be attributable to the activation of Nrf2 pathway via anti-oxidative signaling pathways.") This article is protected by copyright. All rights reserved.
    Keywords:  Icariin; Nerve damage; Nrf2; neurodegenerative diseases; neuroprotection; oxidative stress
    DOI:  https://doi.org/10.1002/bab.1743
  35. Plant Physiol Biochem. 2019 Mar 12. pii: S0981-9428(19)30092-0. [Epub ahead of print]139 92-101
    Chung IM, Venkidasamy B, Thiruvengadam M.
      Nickel oxide nanoparticles (NiO NPs) are utilized in various industries and their release into the environment may lead to the pollution of agricultural areas. However, assessing the toxicity of NiO NPs in major food crops is difficult due to the limited information available on their toxicity. The present investigation was carried out to evaluate how NiO NPs affect plant growth, photosynthetic efficiency, and phytochemical content, as well as changes at the transcriptional level of these phytochemicals in Chinese cabbage seedlings. Chlorophyll, carotenoid, and sugar contents were reduced, while proline and the anthocyanins were significantly upregulated in NiO NPs-treated seedlings. The levels of malondialdehyde, hydrogen peroxide, and reactive oxygen species, as well as peroxidase (POD) enzyme activity, were all enhanced in seedlings exposed to NiO NPs. The levels of glucosinolates and phenolic compounds were also significantly increased in NiO NPs-treated seedlings compared to control seedlings. The expression of genes related to oxidative stress (CAT, POD, and GST), MYB transcription factors (BrMYB28, BrMYB29, BrMYB34, and BrMYB51), and phenolic compounds (ANS, PAP1, and PAL) were significantly upregulated. We suggest that NiO NPs application stimulates toxic effects and enhances the levels of phytochemicals (glucosinolates and phenolic compounds) in Chinese cabbage seedlings.
    Keywords:  Chinese cabbage; Gene expression; Glucosinolates; Nickel oxide nanoparticles; Phenolic compounds; Reactive oxygen species
    DOI:  https://doi.org/10.1016/j.plaphy.2019.03.010
  36. Oncol Rep. 2019 Mar 15.
    Bright SA, Byrne AJ, Vandenberghe E, Browne PV, Mcelligott AM, Meegan MJ, Williams DC.
      The nitrostyrene scaffold was previously identified as a lead target structure for the development of effective compounds targeting Burkitt's lymphoma. The present study aimed to develop these compounds further in haematological malignancies, including chronic lymphocytic leukaemia (CLL). Cellular viability, flow cytometry and lactate dehydrogenase assays, amongst others, were used to examine the effects of nitrostyrene compounds on CLL cells, including a cell line representing disease with poor prognosis (HG‑3) and in ex vivo CLL patient samples (n=14). The results demonstrated that two representative nitrostyrene compounds potently induced apoptosis in CLL cells. The pro‑apoptotic effects of the compounds were found to be reactive oxygen species and caspase‑dependent, and had minimal effects on the viability of normal donor peripheral blood mononuclear cells. Nitrostyrene compounds exhibited synergistic augmentation of apoptosis when combined with the phosphatidylinositol 3‑kinase inhibitor idelalisib and demonstrated potent toxicity in ex vivo CLL cells, including those co‑cultured with bone marrow stromal cells, making them more resistant to apoptosis (n=8). These compounds also demonstrated activity in samples from patients with poor prognostic indicators; unmutated immunoglobulin heavy chain genes, expression of CD38 and deletions in chromosomes 17p and 11q. These results suggest that this class of pharmaceutically active compounds offer potential in the treatment of CLL.
    DOI:  https://doi.org/10.3892/or.2019.7068
  37. Onco Targets Ther. 2019 ;12 1867-1880
    Wang Y, Zhang J, Yang Y, Liu Q, Xu G, Zhang R, Pang Q.
      Purpose: Chemotherapy after surgery can prolong the survival of patients with gliomas. Dimethylaminomicheliolide (DMAMCL), a novel chemotherapeutic agent, exhibited antitumor properties in acute myeloid leukemia stem cells and showed an increased drug concentration in the brain. This study aims to investigate the specific anticancer activities and mechanisms of DMAMCL in glioma cells.Materials and methods: In this study, the effects of DMAMCL were evaluated and characterized in U87-MG and U251 glioma cells. Cell viability was assessed by Cell Counting Kit-8. Apoptosis, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) generation were assessed by fluorescence microscopy. Autophagosome formation was observed with transmission electron microscopy, and the autophagy flux was measured by transfecting cells with mRFP-GFP-LC3 adenoviral vectors. Immunofluorescence and Western blot analyses were used to determine the expression of proteins.
    Results: In the present study, treatment with DMAMCL decreased cell viability and induced apoptosis in U87-MG and U251 glioma cells. Additionally, DMAMCL activated autophagy-mediated cell death as evidenced by the formation of autophagosomes, accumulation of LC3B-II, inhibition of autophagy flux, and increase in cell viability after cotreatment with an autophagy inhibitor. Subsequent experiments showed that the DMAMCL-induced apoptosis and autophagy were possibly mediated by ROS generation and Akt/mTOR signaling pathway inhibition. On the other hand, the ROS scavenger N-acetyl-L-cysteine and the Akt activator insulin-like growth factor-1 attenuated the DMAMCL-induced autophagy and cell death.
    Conclusion: Our findings revealed that DMAMCL induced apoptosis and autophagic cell death by regulating the ROS/mitogen-activated protein kinase signaling pathway and suppressing the Akt/mTOR signaling pathway in human glioma cells. DMAMCL may be a novel effective anticancer agent, which can target gliomas.
    Keywords:  DMAMCL; ROS; apoptosis; autophagy; glioma
    DOI:  https://doi.org/10.2147/OTT.S195329
  38. Environ Sci Pollut Res Int. 2019 Mar 16.
    Rezazadeh Azari M, Mohammadian Y, Pourahmad J, Khodagholi F, Peirovi H, Mehrabi Y, Omidi M, Rafieepour A.
      Co-exposure to carboxylic acid functionalized multi-walled carbon nanotubes (F-MWCNTs) and polycyclic aromatic hydrocarbons (PAHs) such as benzo a pyrene (BaP) in ambient air have been reported. Adsorption of BaP to F-MWCNTs can influence combined toxicity. Studying individual toxicity of F-MWCNTs and BaP might give unrealistic data. Limited information is available on the combined toxicity of F-MWCNTs and BaP in human cells. The objective of the present work is to evaluate the toxicity of F-MWCNTs and BaP individually and combined in human lung adenocarcinoma (A549 cells). The in vitro toxicity is evaluated through cell viability, the production of reactive oxygen species (ROS), apoptosis, and the production of 8-OHdG assays. Adsorption of BaP to F-MWCNTs was confirmed using a spectrophotometer. The results indicated that the F-MWCNTs and BaP reduce cell viability individually and produce ROS, apoptosis, and 8-OHdG in exposed cells. Stress oxidative is found to be a mechanism of cytotoxicity for both F-MWCNTs and BaP. Combined exposure to F-MWCNTs and BaP decreases cytotoxicity compared to individual exposure, but the difference is not statistically significant in all toxicity assays; hence, the two-factorial analysis indicated an additive toxic interaction. Adsorption of BaP to F-MWCNTs could mitigate the bioavailability and toxicity of BaP in biological systems. Considering the mixture toxicity of MWCNTs and BaP is required for risk assessment of ambient air contaminants.
    Keywords:  A549 cells; Benzo a pyrene; Combined toxicity; Functionalized multi-walled carbon nanotubes
    DOI:  https://doi.org/10.1007/s11356-019-04795-x
  39. Physiol Int. 2019 Mar 19. 1-10
    Korkmaz HI, Hahn NE, Jansen KM, Musters R, van Bezu J, van Wieringen WN, van Zuijlen P, Ulrich M, Niessen H, Krijnen P.
      PURPOSE: We previously found that homocysteine (Hcy)-induced apoptosis in endothelial cells coincided with increased NADPH oxidase (NOX) activity. In addition, in ischemic endothelial cells present in the heart, we showed that loss of serine protease dipeptidyl peptidase IV (DPP4) expression was correlated with induction of tissue factor (TF) expression. Since Hcy can initiate thrombosis through the induction of TF expression, in this study, we evaluated whether the inverse relation of TF and DPP4 is also Hcy-dependent and whether NOX-mediated reactive oxygen species (ROS) is playing a role herein.METHODS: Human umbilical vein endothelial cells (HUVECs) were incubated with 2.5 mM Hcy for 3 and 6 h. The effects of Hcy on DPP4 and TF expression and NOX2/p47phox-mediated nitrotyrosine (ROS) production were studied using digital-imaging microscopy.
    RESULTS: In HUVECs, high levels of Hcy showed a significant increase of TF expression and a concomitant loss of DPP4 expression after 6 h. In addition, NOX subunits NOX2 and p47phox were also significantly increased after 6 h of Hcy incubation and coincided with nitrotyrosine (ROS) expression. Interestingly, inhibition of NOX-mediated nitrotyrosine (ROS) with the use of apocynin not only reduced these effects, but also counteracted the effects of Hcy on TF and DPP4 expression.
    CONCLUSION: These results indicate that the inverse relation of TF and DPP4 in endothelial cells is also Hcy-dependent and related to NOX activity.
    Keywords:  DPP4; NADPH oxidase; endothelial cells; homocysteine; tissue factor
    DOI:  https://doi.org/10.1556/2060.106.2019.05
  40. J Vasc Res. 2019 Mar 15. 56(1): 17-27
    Xu W, Hu X, Qi X, Zhu R, Li C, Zhu Y, Yin S, Cheng L, Zhu R.
      Vitamin D has an important protective effect on chronic inflammatory disease. Angiotensin II (AngII) triggers vascular damage and plays a key role in vascular diseases via several mechanisms, including inflammation. Conversely, vitamin D has been shown to have an important protective effect on chronic inflammation. There is evidence showing that vitamin D can reverse the effects of AngII, but the molecular mechanisms by which this occurs are not known. Our results demonstrate that vitamin D improved the viability, migration ability, and tube formation of AngII-pretreated endothelial progenitor cells (EPCs) and inhibited the apoptosis of EPCs induced by AngII. Vitamin D also reversed reactive oxygen species production, vascular inflammatory cytokine generation, and nuclear factor kappa-B activation in EPCs induced by AngII. Furthermore, EPC pretreatment with GW9662 (the antagonist for PPAR-γ) or siHO-1 decreased the protective effect of vitamin D on AngII-induced EPC injury. Overall, our data indicate that vitamin D ameliorated AngII-induced abnormal EPC injury by decreasing oxidative stress and inflammatory cytokine levels. These findings also suggest that vitamin D protected EPCs from AngII-induced vascular injury via the activation of the PPAR-γ/HO-1 signaling pathway.
    Keywords:  Angiotensin II; Endothelial progenitor cells; Heme oxygenase-1; Peroxisome proliferator-activated receptor-gamma; Vascular injury; Vitamin D
    DOI:  https://doi.org/10.1159/000496164
  41. Fish Shellfish Immunol. 2019 Mar 14. pii: S1050-4648(19)30129-9. [Epub ahead of print]88 518-527
    Yang SS, Yu CB, Luo Z, Luo WL, Zhang J, Xu JX, Xu WN.
      The objective of this work was to investigate the effect of berberine (BBR) on the Cell viability, lipid accumulation, apoptosis, cytochrome c, caspase-9 and caspase-3 in lipid accumulation-hepatocytes induced by sodium palmitate in vitro. The lipid accumulation-hepatocytes (induced by 0.5 mM sodium palmitate for 24 h) were treated with 5 μM berberine for 12 h. Then, the Cell viability, intracellular triglyceride (TG) content, lipid peroxide (LPO), malonaldehyde (MDA) content, cytochrome c, caspase-9, caspase-3 and apoptosis were detected. Sodium palmitate decreased Cell viability and increased intracellular TG content, lipid droplet accumulation, LPO and MDA concentrations, caused caspase-3 and caspase-9 activation, then led to apoptosis accompanied by cytochrome c release from mitochondria into the cytoplasm. Beberine could improve intracellular lipid droplet accumulation and oxidative stress, while reduce apoptosis induced by sodium palmitate.
    Keywords:  Apoptosis; Berberine; Hepatocyte; Lipid accumulation; Oxidative stress; Sodium palmitate
    DOI:  https://doi.org/10.1016/j.fsi.2019.02.055
  42. Oxid Med Cell Longev. 2019 ;2019 6505373
    Li C, Li L, Chen K, Wang Y, Yang F, Wang G.
      In recent studies, UFL1 (ubiquitin-like modifier 1 ligating enzyme 1) has been identified as a significant regulator of NF-κB signaling and cellular stress response, yet its physiological function in LPS-stimulated bovine mammary epithelial cells (BMECs) remains unknown. In this study, we investigated the modulating effect of UFL1 on the regulation of LPS-induced inflammation and cell damage, with a focus on apoptosis, ER stress, autophagy, oxidative stress, and the TLR4/NF-κB signaling pathway. The results showed that UFL1 depletion aggravated the LPS-induced inflammatory response and cell damage by positively regulating the TLR4/NF-κB pathway (increased the expression of TLR4, NF-κB P65 in nuclear, and phospho-IκBα), exacerbating LPS-induced ER stress (increased the expression of CHOP, Hsp70, and GRP78), apoptosis (increased the expression of Bax/Bcl-2 and activity of caspase-3), autophagy (increased LC3-II and decreased P62 expression), and oxidative stress (decreased SOD and CAT levels and increased MDA levels). Overexpression of UFL1 suppressed the activation of the TLR4/NF-κB pathway and relieved the LPS-induced ER stress, apoptosis, autophagy, and oxidative stress, thereby alleviating the inflammatory response and cell damage. Collectively, UFL1 may play an important role during the inflammatory response and thereby acts as a potential therapeutic target for bovine mastitis.
    DOI:  https://doi.org/10.1155/2019/6505373
  43. Mol Nutr Food Res. 2019 Mar 20. e1801353
    Ou HC, Chou WC, Chu PM, Hsieh PL, Hung CH, Tsai KL.
      SCOPE: Atherosclerotic cardiovascular disease is the most prevalent cause of mortality and morbidity. Fucoxanthin (FX) possesses anti-hypertensive and anti-obesity properties. However, the molecular mechanisms underlying the inhibitory effects of FX on oxidized low-density lipoprotein (oxLDL)-induced oxidative injuries in human endothelial cells are still largely unknown. This study aimed to test the hypothesis that FX protects against oxLDL-induced oxidative stress by upregulating AMP-activated protein kinase (AMPK) and to explore the roles of cAMP response element binding protein (CREB) and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α).METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were treated with oxLDL in the presence or absence of FX. FX significantly increased AMPK phosphorylation. In addition, FX diminished oxLDL-mediated nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation by inhibiting protein kinase C (PKC) and subsequently inducing reactive oxygen species (ROS) generation and impairing the activity of the endogenous antioxidant enzyme superoxidase dismutase (SOD). Furthermore, FX restored oxLDL-mediated dephosphorylation of phosphoinositide-3-kinase/Akt (PI3K/Akt) and decreased CREB and PGC-1α expression to nearly normal levels. Moreover, FX ameliorated the oxLDL-mediated suppression of mitochondrial function and apoptosis.
    CONCLUSION: These findings provide new insights into the possible molecular mechanisms by which FX mitigates oxLDL-induced endothelial oxidative stress and mitochondrial dysfunction. This article is protected by copyright. All rights reserved.
    Keywords:  AKT; AMPK; Fucoxanthin; endothelial dysfunction; oxidative stress
    DOI:  https://doi.org/10.1002/mnfr.201801353
  44. Curr Drug Targets. 2019 Mar 19.
    Hosseinzadeh A, Kamrava SK, Moore BCJ, Reiter RJ, Ghaznavi H, Kamali M, Mehrzadi S.
      Tinnitus is a hearing disorder characterized by the perception of sound without external acoustic stimuli, which is caused by damage to the auditory system in response to excessive levels of noise, ototoxic agents and aging process. Neural plasticity, oxidative/nitrosative stress and apoptosis play important roles in the pathogenesis of tinnitus. The expression of neural plasticity related to excessive glutamatergic neurotransmission leads to generation of abnormal sound in one's ears or head. Furthermore, hyperactivation and over-expression of NMDA receptors in response to excessive release of glutamate contribute to the calcium overload in the primary auditory neurons and subsequent cytotoxicity. Reactive oxygen/nitrogen species are endogenously produced by different type of cochlear cells under pathological conditions, which cause direct damage to the intracellular components and apoptotic cell death. Cochlear hair-cell death contributes to the progressive deafferentation of auditory neurons, which consequently leads to the aberrant activity in several parts of the auditory pathway. Therefore, targeting neural plasticity, oxidative/nitrosative stress, apoptosis and autophagy may ameliorate ringing in the ear. Melatonin is an endogenously produced indoleamine synchronizing circadian and circannual rhythms. Based on laboratory studies indicating the protective effect of melatonin against cochlear damage induced by acoustic trauma and ototoxic agents, and also clinical studies reporting the ability of melatonin to minimize the severity of tinnitus, melatonin is suggested to be a treatment option for patient with buzzing sound in the ear. Herein, we describe the ameliorative effect of melatonin on tinnitus focusing on neural plasticity, oxidative/nitrosative stress, apoptotsis and autophagy.
    Keywords:  Apoptosis; Autophagy; Melatonin; Neural plasticity; Oxidative stress; Tinnitus
    DOI:  https://doi.org/10.2174/1389450120666190319162147
  45. Int J Oncol. 2019 Mar 14.
    Fan JJ, Hsu WH, Hung HH, Zhang WJ, Lee YA, Chen KC, Chu CY, Ko TP, Lee MT, Lin CW, Cheng CH.
      Reactive oxygen species (ROS) homeostasis is maintained at a higher level in cancer cells, which promotes tumorigenesis. Oxidative stress induced by anticancer drugs may further increase ROS to promote apoptosis, but can also enhance the metastasis of cancer cells. The effects of ROS homeostasis on cancer cells remain to be fully elucidated. In the present study, the effect of a reduction in manganese superoxide dismutase (MnSOD) on the migration and invasion of A431 cells was investigated. Our previous micro‑assay data revealed that the mRNA expression of MnSOD was higher in the invasive A431‑III cell line compared with that in the parental A431 cell line (A431‑P). In the present study, high protein levels of MnSOD and H2O2 production were observed in A431‑III cells; however, catalase protein levels were significantly lower in A431‑III cells compared with those in the A431‑P cell line. The knockdown of MnSOD increased H2O2 levels, enzyme activity, the mRNA levels of matrix metalloproteinase‑1, ‑2 and ‑9, and the migratory and invasive abilities of the cells. Inducing a reduction in H2O2 using diphenyleneiodonium (DPI) and N‑acetyl‑l‑cysteine decreased the migratory abilities of the cell lines, and DPI attenuated the migratory ability that had been increased by MnSOD small interfering RNA knockdown. Luteolin (Lu) and quercetin (Qu) increased the expression of catalase and reduced H2O2 levels, but without an observed change in the protein levels of MnSOD. Taken together, these data suggest that reduced MnSOD may induce ROS imbalance in cells and promote the metastatic ability of cancer cells. Lu and Qu may attenuate these processes and may be promising potential anticancer agents.
    DOI:  https://doi.org/10.3892/ijo.2019.4750
  46. Mol Biol (Mosk). 2019 Jan-Feb;53(1):53(1): 84-90
    Mukhammadiyeva GF, Karimov DO, Kutlina TG, Valova YV, Khusnutdinova NY, Repina EF, Bakirov AB.
      Carbon tetrachloride is a well-studied hepatotropic poison. Animal models of exposure to carbon tetrachloride resemble acute liver damage in humans. This paper presents the study of the expression of genes related to cell cycle control, apoptosis, and oxidative stress in a model of carbon tetrachloride-induced toxic hepatitis in rats. White mongrel male rats were injected with a 50% oil solution of carbon tetrachloride at a dose of 0.125-4.000 g/kg (experimental group) or olive oil (control group). The animals were decapitated 24 and 72 h after the administration of carbon tetrachloride, and the qRT-PCR expression levels of the genes encoding hemoxygenase-1 (Hmox1), cell cycle checkpoint kinase-1 (Chek1), and caspase-7 (Casp7) in the liver were analyzed. The increase in the expression levels of Hmox1 and Chek1 after exposure was detected. These genes may either play a role in promoting pathological oxidative stress in the liver or be a part of a stress response. We have concluded that the major pathway of the liver damage in carbon tetrachloride exposed animals is necrosis rather than apoptosis.
    Keywords:  Casp7; Chek1; Hmox1; carbon tetrachloride; gene expression; oxidative stress; toxic liver damage
    DOI:  https://doi.org/10.1134/S0026898419010105
  47. J Adv Res. 2019 Mar;16 87-97
    Sani D, Khatab NIO, Kirby BP, Yong A, Hasan S, Basri H, Stanslas J.
      Substantial evidence has shown that most cases of memory impairment are associated with increased neuroinflammation and oxidative stress. In this study, the potential of a standardised Andrographis paniculata aqueous extract (APAE) to reverse neuroinflammation and cognitive impairment induced by lipopolysaccharide (LPS) was examined in vivo. Rats were treated with APAE (50, 100, 200, and 400 mg·kg-1, p.o.) for 7 consecutive days prior to LPS (1 mg·kg-1, i.p.)-induced neuroinflammation and cognitive impairment. Spatial learning and memory were evaluated using the Morris water maze (MWM) test, while neuroinflammation and oxidative stress were assessed through the measurement of specific mediators, namely, tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, superoxide dismutase (SOD), catalase (CAT), antioxidant glutathione (GSH), reactive oxygen species (ROS), and thiobarbituric acid reactive substance (TBARS). Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) were also evaluated. LPS caused significant memory deficits in the 2-day MWM protocol, whereas pretreatment with standardised APAE dose-dependently improved performance in the MWM test. APAE treatment also blocked the LPS-induced hippocampal increase in the concentration and expression of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and production of ROS and TBARS and enhanced the activities of AChE and BChE. Furthermore, APAE enhanced the decrease in the levels and expression of hippocampal antioxidant enzymes (SOD and CAT) following LPS-induced neuroinflammation and cognitive deficit. The findings from these studies suggested that standardised APAE improved memory and had potent neuroprotective effects against LPS-induced neurotoxicity.
    Keywords:  Cognitive deficits; LPS; MWM; Neuroinflammation; Spatial learning and memory; Standardised APAE
    DOI:  https://doi.org/10.1016/j.jare.2018.11.005
  48. J Dermatolog Treat. 2019 Mar 21. 1-27
    Yuan L, Duan X, Zhang R, Zhang Y, Qu M.
      The aim of this study was to investigate if aloe polysaccharide (AP) has the repairing effect on ultraviolet b (UVB) injured nerve cells. The study applied BALB/c female mice as animal model, and NFG activated PC12 cells as cell model of skin nerve. The cell viability was detected by MTT assay, and cell apoptosis was detected by TUNEL (TdT-mediated dUTP nick-end labeling) and Annexin-/PI assay, and cell cycle status in different groups were observed via flow cytometry (FCM). Enzyme-linked immunosorbent assay (ELISA) was applied to analyze oxidative stress and anti-oxidative ability in each group. Real-time PCR and western blot were used to detect the expression levels of Bax, Bcl-2, Caspase-3, Cyclin D1, Keap1, Nrf2, GCLC and GSTP1. The results showed obvious inhibition of cell viability and cell cycle progression and promotion of cell apoptosis by UVB irradiation through inducing oxidative stress. In AP treated groups, cell viability and proliferation could be markedly improved and cell apoptosis inhibited with higher anti-oxidative capability and up-regulated expression of Keap1, Nrf2, GCLC and GSTP1. It suggested that AP was able to repair UVB induced injury on NGF activated skin neural cell PC12, probably through Keap1/Nrf2/ARE signal pathway.
    Keywords:  Keap1; Nrf2; PC12 cells; Ultraviolet b; ale polysaccharide; antioxidant
    DOI:  https://doi.org/10.1080/09546634.2019.1591579
  49. Oxid Med Cell Longev. 2019 ;2019 9783429
    Bungau S, Abdel-Daim MM, Tit DM, Ghanem E, Sato S, Maruyama-Inoue M, Yamane S, Kadonosono K.
      Oxidative stress and inflammation play a critical role in the initiation and progression of age-related ocular abnormalities as cataract, glaucoma, diabetic retinopathy, and macular degeneration. Therefore, phytochemicals with proven antioxidant and anti-inflammatory activities, such as carotenoids and polyphenols, could be of benefit in these diseases. We searched PubMed and Web of Science databases for original studies investigating the benefits of different carotenoids and polyphenols in age-related ophthalmic diseases. Our results showed that several polyphenols (such as anthocyanins, Ginkgo biloba, quercetin, and resveratrol) and carotenoids (such as lutein, zeaxanthin, and mezoxanthin) have shown significant preventive and therapeutic benefits against the aforementioned conditions. The involved mechanisms in these findings include mitigating the production of reactive oxygen species, inhibiting the tumor necrosis factor-α and vascular endothelial growth factor pathways, suppressing p53-dependent apoptosis, and suppressing the production of inflammatory markers, such as interleukin- (IL-) 8, IL-6, IL-1a, and endothelial leucocyte adhesion molecule-1. Consumption of products containing these phytochemicals may be protective against these diseases; however, adequate human data are lacking. This review discusses the role and mechanisms of polyphenols and carotenoids and their possible synergistic effects on the prevention and treatment of age-related eye diseases that are induced or augmented by oxidative stress and inflammation.
    DOI:  https://doi.org/10.1155/2019/9783429
  50. Int J Nanomedicine. 2019 ;14 1469-1487
    Liao S, Zhang Y, Pan X, Zhu F, Jiang C, Liu Q, Cheng Z, Dai G, Wu G, Wang L, Chen L.
      Background: The threat of drug-resistant Pseudomonas aeruginosa requires great efforts to develop highly effective and safe bactericide.Objective: This study aimed to investigate the antibacterial activity and mechanism of silver nanoparticles (AgNPs) against multidrug-resistant P. aeruginosa.
    Methods: The antimicrobial effect of AgNPs on clinical isolates of resistant P. aeruginosa was assessed by minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). In multidrug-resistant P. aeruginosa, the alterations of morphology and structure were observed by the transmission electron microscopy (TEM); the differentially expressed proteins were analyzed by quantitative proteomics; the production of reactive oxygen species (ROS) was assayed by H2DCF-DA staining; the activity of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) was chemically measured and the apoptosis-like effect was determined by flow cytometry.
    Results: Antimicrobial tests revealed that AgNPs had highly bactericidal effect on the drug-resistant or multidrug-resistant P. aeruginosa with the MIC range of 1.406-5.625 µg/mL and the MBC range of 2.813-5.625 µg/mL. TEM showed that AgNPs could enter the multidrug-resistant bacteria and impair their morphology and structure. The proteomics quantified that, in the AgNP-treated bacteria, the levels of SOD, CAT, and POD, such as alkyl hydroperoxide reductase and organic hydroperoxide resistance protein, were obviously high, as well as the significant upregulation of low oxygen regulatory oxidases, including cbb3-type cytochrome c oxidase subunit P2, N2, and O2. Further results confirmed the excessive production of ROS. The antioxidants, reduced glutathione and ascorbic acid, partially antagonized the antibacterial action of AgNPs. The apoptosis-like rate of AgNP-treated bacteria was remarkably higher than that of the untreated bacteria (P<0.01).
    Conclusion: This study proved that AgNPs could play antimicrobial roles on the multidrug-resistant P. aeruginosa in a concentration- and time-dependent manner. The main mechanism involves the disequilibrium of oxidation and antioxidation processes and the failure to eliminate the excessive ROS.
    Keywords:  AgNPs; Pseudomonas aeruginosa; antibacterial activity; mechanism; multidrug-resistant bacterium; silver nanoparticles
    DOI:  https://doi.org/10.2147/IJN.S191340
  51. Int J Nanomedicine. 2019 ;14 1597-1618
    Zhao C, Cao W, Zheng H, Xiao Z, Hu J, Yang L, Chen M, Liang G, Zheng S, Zhao C.
      Objective: Nanoparticles can efficiently carry and deliver anticancer agents to tumor sites. Mounting evidence indicates that many types of cancer cells, including colon cancer, have a weakly acidic microenvironment and increased levels of reactive oxygen species. The construction of nano drug delivery vehicles "activatable" in response to the tumor microenvironment is a new antitumor therapeutic strategy.Methods: Cinnamaldehyde (CA) was designed to link directly with dextran to form a polymer through an acid cleavable acetal bond. Herein, a novel pH-sensitive drug delivery system was constructed with co-encapsulated 10-hydroxy camptothecin (HCPT). Dynamic light scattering (DLS) analysis, transmission electron microscopy (TEM) analysis, and release kinetics analysis of HCPT-CA-loaded nanoparticles (PCH) were conducted to investigate the physical and chemical properties. The cellular uptake signatures of the nanoparticles were observed by confocal microscopy and flow cytometry. Cell viability, cell scratch assay, apoptosis assay, and colony formation assay were performed to examine the potent antiproliferative and apoptotic effects of the PCH. The antitumor mechanism of the treatment with PCH was evaluated by Western blotting, flow cytometry, and TEM analysis. The pharmacokinetics of PCH were examined in healthy Sprague Dawley rats within 6 hours after sublingual vein injection. We lastly examined the biodistribution and the in vivo anticancer activity of PCH using the xenograft mouse models of HCT116 cells.
    Results: Both HCPT and CA were quickly released by PCH in an acidic microenvironment. PCH not only induced cancer cell death through the generation of intracellular reactive oxygen species in vitro but also facilitated the drug uptake, effectively prolonged drug circulation, and increased accumulation of drug in tumor sites. More attractively, PCH exhibited excellent therapeutic performance and better in vivo systemic safety.
    Conclusion: Overall, PCH not only utilized the tumor microenvironment to control drug release, improve drug pharmacokinetics, and passively target the drug to the tumor tissue, but also exerted a synergistic anticancer effect. The acid-responsive PCH has enormous potential as a novel anticancer therapeutic strategy.
    Keywords:  ROS; cinnamaldehyde; colon cancer; hydroxy camptothecin; pH-responsive nanoparticles
    DOI:  https://doi.org/10.2147/IJN.S189923
  52. Biochem Biophys Res Commun. 2019 Mar 19. pii: S0006-291X(19)30459-0. [Epub ahead of print]
    Liu Y, Huang X, Wang P, Pan Y, Cao D, Liu C, Chen A.
      Skin photoaging refers to the phenomenon of skin aging or accelerated aging as a result of long-term UV exposure. Ultraviolet radiation can lead to DNA damage, cell apoptosis, cell growth inhibition and carcinogenic effects. Evidence suggests that hsp27 can protect cells from apoptosis induced by various stimuli in vivo and in vitro. However, modulation in hsp27 expression toward skin protection against UVB treatment has not been investigated clearly. In this study, we aimed to investigate the effects of hsp27 against UVB-induced photoaging in rat skin and to explore the underlying mechanisms. In the present study, we identified that the level of hsp27 increased after UVB irradiation induced chronic photoaging rat model. In order to investigate the function of hsp27 in UVB-induced skin photoaging, we used adeno-associated virus (AAV) to specificity reduce the expression of hsp27 in rat skin. In contrast to UVB group, we found that collagen fibers were disorganized and elastic fibers were thickened and twisted in UVB-AAV group. In the UVB-AAV group, reduced hsp27 enhanced the oxidative stress. Aging markers (SA-β-Gal staining and the protein levels of p16, p53, p21) were significantly changed in the hsp27 decreased group. However, in hsp27 deletion group, the expression of antiapoptotic factor bcl-2 was decreased, while the apoptosis factor bax was increased after UVB irradiation. These findings suggested that hsp27 was involved in oxidative stress, aging and apoptosis of skin after UV exposure. Management the expression of hsp27 can be used as a potential intervention method to alleviate UVB-induced skin damage.
    Keywords:  Apoptosis; HSP27 expression; Photoaging; Ultraviolet radiation
    DOI:  https://doi.org/10.1016/j.bbrc.2019.03.076
  53. Toxicol Mech Methods. 2019 Mar 20. 1-37
    Li J, Zheng L, Wang X, Yao K, Shi L, Sun X, Yang G, Jiang L, Zhang C, Wang Y, Jiang L, Liu X.
      Di-(2-ethylhexyl) phthalate (DEHP) is an environmental endocrine disruptor widely employed in plastic bags, industrial paints, cosmetics and food packaging, which has been reported to be harmful to human physical health. Many studies have shown that DEHP causes reproductive system toxicity, but its cytotoxicity to islet cells is few to unknown. In our research, it was found that DEHP could induce apoptosis in INS-1 cells via autophagy and oxidative stress. Taurine, a sulfur-containing β-amino acid, could reverse DEHP-induced oxidative stress imbalance. Meanwhile, taurine could reduce DEHP-induced excessive autophagy. The interaction between oxidative stress and autophagy has been investigated in this study. After pretreated with autophagy interventional agents, it was found that autophagy was capable of alleviating oxidative stress and ROS production in DEHP-treated INS-1 cells. And down-regulated ROS production by NAC could also turn over uploaded autophagy. Our research provides a perspective about the mechanism of cytotoxicity of DEHP to INS-1 cells and taurine protective effect.
    Keywords:  DEHP; INS-1 cells; apoptosis; autophagy; oxidative stress
    DOI:  https://doi.org/10.1080/15376516.2019.1588931
  54. Oxid Med Cell Longev. 2019 ;2019 1701478
    Avdonin PV, Nadeev AD, Mironova GY, Zharkikh IL, Avdonin PP, Goncharov NV.
      Hydrogen peroxide, formed in the endothelium, acts as a factor contributing to the relaxation of blood vessels. The reason for this vasodilatory effect could be modulation by H2O2 of calcium metabolism, since mobilization of calcium ions in endothelial cells is a trigger of endothelium-dependent relaxation. The aim of this work was to investigate the influence of H2O2 on the effects of Ca2+-mobilizing agonists in human umbilical vein endothelial cells (HUVEC). We have found that H2O2 in concentration range 10-100 μM increases the rise of [Ca2+]i induced by 5-hydroxytryptamine (5-HT) and carbachol and does not affect the calcium signals of ATP, agonist of type 1 protease-activated receptor SFLLRN, histamine and bradykinin. Using specific agonists of 5-HT1B and 5-HT2B receptors CGS12066B and BW723C86, we have demonstrated that H2O2 potentiates the effects mediated by these types of 5-HT receptors. Potentiation of the effect of BW723C86 can be produced by the induction of endogenous oxidative stress in HUVEC. We have shown that the activation of 5-HT2B receptor by BW723C86 causes production of reactive oxygen species (ROS). Inhibitor of NADPH oxidases VAS2870 suppressed formation of ROS and partially inhibited [Ca2+]i rise induced by BW723C86. Thus, it can be assumed that vasorelaxation induced by endogenous H2O2 in endothelial cells partially occurs due to the potentiation of the agonist-induced calcium signaling.
    DOI:  https://doi.org/10.1155/2019/1701478
  55. Sci Rep. 2019 Mar 20. 9(1): 4954
    Huang H, Aladelokun O, Ideta T, Giardina C, Ellis LM, Rosenberg DW.
      The platinum-based chemotherapeutic agent, oxaliplatin, is used to treat advanced colorectal cancer (CRC). Unfortunately, nearly all patients acquire resistance to oxaliplatin after long-term use, limiting its therapeutic efficacy. Since COX-2 and PGE2 signaling can impact colon cancer cell proliferation and survival, we examined how this pathway was affected in an oxaliplatin resistant colon cancer cell line. PGE2 levels were significantly elevated in oxaliplatin-resistant HT29 cells (OXR) compared to naïve parental HT29 cells (PAR). This increase was associated with elevated COX-2 (17.9-fold; P = 0.008) and reduced 15-hydroxyprostaglandin dehydrogenase (2.9-fold; P < 0.0001) expression. RNAi knockdown of microsomal prostaglandin E synthase-1, the rate-limiting enzyme in PGE2 synthesis, sensitized OXR cells to oxaliplatin. Downstream effects of PGE2 in OXR cells were also examined. Selective inhibition of the EP4 PGE2 receptor by the small molecule inhibitor, L-161,982 enhanced oxaliplatin-induced apoptosis in OXR cells. L-161,982 also reduced expression of the colonic stem cell markers, CD133 and CD44, and inhibited tumor sphere formation. The accumulation of intracellular reactive oxygen species (ROS), a key component of oxaliplatin cytotoxicity, was significantly increased by EP4 inhibition (2.4 -fold; P < 0.0001). Overall, our findings uncover an important role for the COX-2/PGE2/EP4 signaling axis in oxaliplatin resistance via regulation of oxidative stress.
    DOI:  https://doi.org/10.1038/s41598-019-40848-4
  56. Oxid Med Cell Longev. 2019 ;2019 8039694
    Xue R, Wang J, Yang L, Liu X, Gao Y, Pang Y, Wang Y, Hao J.
      Aim: Pancreatic stellate cells (PSCs) play a pivotal role in pancreatic fibrosis. Any remedies that inhibit the activation of PSCs can be potential candidates for therapeutic strategies in pancreatic fibrosis-related pancreatic ductal adenocarcinoma (PDAC) and chronic pancreatitis (CP). Our study is aimed at exploring the protective effect of coenzyme Q10 (CoQ10) against pancreatic fibrosis.Methods: Pancreatic fibrosis was induced by 20% L-arginine (250 mg/100 g) at 1 h intervals twice per week for 8 weeks in C57BL/6 mice. CoQ10 was administered for 4 weeks. Isolated primary PSCs from C57BL/6 mice were treated with 100 μM CoQ10 for 72 h, as well as Rosup and specific inhibitors. The effects of CoQ10 on the activation of PSCs, autophagy, collagen deposition, histological changes, and oxidative stress were analyzed by western blotting, biochemical estimations, immunofluorescence staining, and hematoxylin-eosin, Masson, and Sirius red staining, as well as with a reactive oxygen species (ROS) assay.
    Results: Pretreatment and posttreatment of CoQ10 decreased autophagy, activation of PSCs, oxidative stress, histological changes, and collagen deposition in the CP mouse model. In primary PSCs, expression levels of p-PI3K, p-AKT, and p-mTOR were upregulated with CoQ10. A rescue experiment using specific inhibitors of the PI3K-AKT-mTOR pathway demonstrated that the PI3K-AKT-mTOR signaling pathway was the underlying mechanism by which CoQ10 ameliorated fibrosis. With the addition of Rosup, expression levels of the autophagy biomarkers LC3 and Atg5 were elevated. Meanwhile, the levels of p-PI3K, p-AKT, and p-mTOR were lower.
    Conclusions: Our findings demonstrated that CoQ10 alleviates pancreatic fibrosis by the ROS-triggered PI3K/AKT/mTOR signaling pathway. CoQ10 may be a therapeutic candidate for antifibrotic methods.
    DOI:  https://doi.org/10.1155/2019/8039694
  57. Eur J Med Chem. 2019 Mar 09. pii: S0223-5234(19)30211-9. [Epub ahead of print]170 45-54
    Yang K, Li Y, Tang Q, Zheng L, He D.
      Mitochondria are considered as a therapeutic target for new drug design toward all kinds of cancer. Hence in order to enhance the dosage in mitochondrial fraction of cinnamamides, the mitochondria-targeted derivatives were designed by the incorporation of cinnamamides into a fluorophore carrier of coumarin-3-carboxamide with a 1:1 stoichiometry. Using the amide linkers, twenty-one compounds were synthesized and the cytotoxicity against a panel of cancer cells (MCF-7, Hela, HepG2, HL-60) was tested. In particular, compound 18c displayed the potent cytotoxicity toward HL-60 leukaemia cells, which could quickly and efficiently entry into HL-60 cells and specifically localize within mitochondria. And 18c preferred enrichment in HL-60 cells than in PBMC normal cells, accounting for the higher toxicity to cancer cells than to normal cells. Moreover, the dissipations of mitochondrial membrane potential and enhancement of cellular ROS level were also preceded upon 18c treatment, leading to cell cycle arrest and apoptosis/necrosis in HL-60 cells. Besides, acted as a Michael acceptor, 18c initiated a thia-Michael addition reaction toward cysteamine (1:2 stoichiometry), detecting by the UV-Vis spectrum and HRMS analysis. This could result in the blue emission of 18c in mitochondria after the procedure of cell fixation, owing to the formation of covalent bond with mitochondrial thiols. Our study reported 18c might be useful for the further development into a mitochondria-targeted anti-leukemia agent and the Michael acceptor might be a versatile functional group.
    Keywords:  Cinnamamide; Cytotoxicity; Fluorescent image; Targeting mitochondria; Thia-Michael addition; Uptake
    DOI:  https://doi.org/10.1016/j.ejmech.2019.03.001
  58. Nanomaterials (Basel). 2019 Mar 18. pii: E451. [Epub ahead of print]9(3):
    Jelinek M, Kocourek T, Jurek K, Jelinek M, Smolková B, Uzhytchak M, Lunov O.
      This paper deals with the synthesis and study of the properties of germanium-doped diamond-like carbon (DLC) films. For deposition of doped DLC films, hybrid laser technology was used. Using two deposition lasers, it was possible to arrange the dopant concentrations by varying the laser repetition rate. Doped films of Ge concentrations from 0 at.% to 12 at.% were prepared on Si (100) and fused silica (FS) substrates at room temperature. Film properties, such as growth rate, roughness, scanning electron microscopy (SEM) morphology, wavelength dependent X-ray spectroscopy (WDS) composition, VIS-near infrared (IR) transmittance, and biological properties (cytotoxicity, effects on cellular morphology, and ability to produce reactive oxygen species (ROS)) were studied in relation to codeposition conditions and dopant concentrations. The analysis showed that Ge-DLC films exhibit cytotoxicity for higher Ge doping.
    Keywords:  DLC; apoptosis; cytotoxicity; doped biomaterials; germanium; pulsed laser deposition; reactive oxygen species
    DOI:  https://doi.org/10.3390/nano9030451
  59. J Cell Physiol. 2019 Mar 19.
    Fu Y, Sun S, Sun H, Peng J, Ma X, Bao L, Ji R, Luo C, Gao C, Zhang X, Jin Y.
      Atherosclerosis (AS), a progressive disorder, is one of the tough challenges in the clinic. Scutellarin, an extract from Herba Erigerontis, is found to have oxygen-free radicals scavenging effects and antioxidant effects. In this study, we aimed to investigate the anti-AS effects of scutellarin is related to controlling the Hippo-FOXO3A and PI3K/AKT signal pathway. To establish an AS model, the rats in the scutellarin and model groups were intraperitoneally injected with vitamin D 3 and then fed a high-fat diet for 12 weeks. In addition, in vitro angiotensin II-induced apoptosis of human aortic endothelial cells (HAECs) were used to establish models. Scutellarin significantly reduced blood lipid levels and increased antioxidase levels in both models. Additionally, scutellarin inhibited reactive oxygen species generation and apoptosis in HAECs. The impaired vascular barrier function was restored by using scutellarin in AS rats and in HAECs cells characterized by inhibiting mammalian sterile-20-like kinases 1 (Mst1) phosphorylation, Yes-associated protein (YAP) phosphorylation, forkhead box O3A (FOXO3A) phosphorylation at serine 207, nuclear translocation of FOXO3A, and upregulating protein expression of AKT and FOXO3A phosphorylation at serine 253. Scutellarin significantly reduced Bcl-2 interacting mediator of cell death (Bim), caspase-3, APO-1, CD95 (Fas), and Bax: Bcl-2-associated X (Bax) levels and activated Bcl-2: B-cell lymphoma-2 (Bcl-2). Scutellarin also significantly inhibited the expression of Mst1, YAP, FOXO3A at the messenger RNA level. When Mst1 was overexpressed or phosphoinositide 3-kinases suppressed, the effects of scutellarin were significantly blocked. In conclusion, the results of the present study suggest that scutellarin exerts protective effects against AS by inhibiting endothelial cell injury and apoptosis by regulating the Hippo-FOXO3A and PI3K/AKT signal pathways.
    Keywords:  Hippo-FOXO3A; PI3K/AKT; antioxidation; apoptosis; atherosclerosis; scutellarin
    DOI:  https://doi.org/10.1002/jcp.28446
  60. Med Oncol. 2019 Mar 22. 36(4): 38
    Zhang J, Song J, Wu D, Wang J, Dong W.
      The original version of this article unfortunately contained a mistake. During final figure preparation and manuscript submission, the similar graphs (Fig. 6c) in different papers were accidentally switched. It was subsequently noted in a further review that Fig. 2a does not match with the hesperetin concentration. The corrected figures and graphs are herein (Figs. 2a, 6c). This correction does not affect the results or conclusions of the original paper.
    DOI:  https://doi.org/10.1007/s12032-019-1258-0
  61. Autophagy. 2019 Mar 20.
    Xian H, Liou YC.
      Mitochondrial dynamics is highly implicated in a plethora of cellular processes including apoptosis and mitophagy. However, little is known about the scope and precise functions of mitochondrial dynamics proteins for mitochondrial quality control and cellular homeostasis. Whether mitochondrial dynamics proteins serve in cellular processes reliant on mitochondrial fission-fusion is still not fully explored. MIEF1/MiD51 (mitochondrial elongation factor 1) is known to promote mitochondrial fission via the recruitment of GTPase protein DNM1L/DRP1 (dynamin 1 like), but the fundamental understandings of MIEF1 for mitochondrial-dependent cellular processes are largely elusive. Here, we report novel roles of MIEF1 in responding to apoptotic stimuli and mitochondrial damage. Given our result that staurosporine (STS) treatment induced the degradation of MIEF1 via the ubiquitin-proteasome system (UPS), we are motivated to explore the role of MIEF1 in apoptosis. MIEF1 loss triggered the imbalance of BCL2 family members on the mitochondria, consequently initiating the translocation of BAX onto the mitochondria, catalyzing the decrease of mitochondrial membrane potential and promoting the release of DIABLO/SMAC (diablo IAP-binding mitochondrial protein) and CYCS (cytochrome c, somatic). We further demonstrate that MIEF1 deficiency impaired mitochondrial respiration and induced mitochondrial oxidative stress, sensitizing cells to PINK1-PRKN-mediated mitophagy. The recruitment of PRKN to depolarized mitochondria modulated the UPS-dependent degradation of MFN2 (mitofusin 2) and FIS1 (fission, mitochondrial 1) specifically, to further promote mitophagy. Our findings uncover a bridging role of MIEF1 integrating cell death and mitophagy, unlikely dependent on mitochondrial dynamics, implying new insights to mechanisms determining cellular fate.
    Keywords:  BAX; MIEF1; PRKN; ROS; apoptosis; mitochondria; mitophagy
    DOI:  https://doi.org/10.1080/15548627.2019.1596494
  62. Onco Targets Ther. 2019 ;12 1729-1739
    Zhou L, Liu L, Chai W, Zhao T, Jin X, Guo X, Han L, Yuan C.
      Background: Metabolic reprogramming is a characteristic of tumor cells and is considered a potential therapeutic target. Even under aerobic conditions, tumor cells use glycolysis to produce energy, a phenomenon called the "Warburg effect". Pyruvate dehydrogenase kinase 1 (PDK1) is a key factor linking glycolysis and the tricarboxylic acid cycle. Dichloroacetic acid (DCA) reverses the Warburg effect by inhibition of PDK1 to switch cytoplasmic glucose metabolism to mitochondrial oxidative phosphorylation (OXPHOS).Methods: Cell viability was examined using a standard MTT assay. Glucose consumption and l-lactate production were measured using commercial colorimetric kits, and intracellular lactate dehydrogenase (LDH) activity was evaluated using cell lysates and an LDH Quantification Kit. Real-time PCR was used to detect the expression of related genes. The production of total ROS was evaluated by staining with dichlorofluorescin diacetate.
    Results: Comparison of various aspects of glucose metabolism, such as expression of key enzymes in glycolysis, lactate production, glucose consumption, mitochondrial oxygen consumption rate, and citric acid production, revealed that A2780/DDP cells were primarily dependent on glycolysis whereas A2780 cells were primarily dependent on mitochondrial OXPHOS. Mitochondrial uncoupling protein 2 (UCP2) protects against mitochondrial ROS while allowing energy metabolism to switch to glycolysis. Treatment of A2780 cells with various concentrations of DCA resulted in decreased expression of UCP2, a metabolic switch from glycolysis to mitochondrial OXPHOS, and an increase in oxidative stress induced by ROS. These effects were not observed in A2780/DDP cells with higher UCP2 expression suggesting that UCP2 might induce changes in mitochondrial functions that result in different sensitivities to DCA.
    Conclusion: Our results show that a drug targeting tumor metabolic changes affects almost the entire process of glucose metabolism. Thus, it is necessary to comprehensively determine tumor metabolic functions to facilitate individualized antitumor therapy.
    Keywords:  DCA; glucose; glycolysis; metabolism; mitochondrial function
    DOI:  https://doi.org/10.2147/OTT.S194329
  63. Arch Med Sci. 2019 Mar;15(2): 467-474
    Kamisli S, Basaran C, Batcioglu K, Oztanir MN, Gul M, Satilmis B, Uyumlu AB, Kayhan B, Genc M.
      Introduction: RS100642, a mexiletine analogue, is a novel sodium channel blocker with neuroprotective and antioxidant activities. The protectivity of RS100642, which has been shown against focal cerebral ischemia, was investigated in global cerebral ischemia in this study.Material and methods: Global cerebral ischemia was induced for five minutes in adult male Wistar Albino rats via the 4-vessel occlusion method. Intravenous administration of 1 mg/kg RS100642 following reperfusion for 30 min (RS100642 group) was compared with a sham treatment group (ischemia group) and nonischemized group (control) histologically based on morphology and caspase-3 immunohistochemistry, and biochemically based both on measurement of oxidative stress including malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) activities and on assessment of apoptosis including caspase-3 and -8 activities and tumor necrosis factor α (TNF-α) levels at the end of 6 h.
    Results: While the RS100642 group had significantly lower MDA levels and higher SOD activities than the sham treatment group (p < 0.05), GPx and CAT activities of the RS100642 and sham treatment groups were similar (p > 0.05) and significantly lower than those of the controls (p < 0.05). Necrosis and caspase-3 activity and immunoreactivity in the RS100642 group were significantly lower than those in the sham treatment group (p < 0.05), while there was no significant difference between groups regarding caspase-8 and TNF-α (p > 0.05).
    Conclusions: Na+ channel blockade by RS100642 has remarkable neuroprotective effects following global brain ischemia/reperfusion damage. Further research is required to determine the optimum dose and time of administration.
    Keywords:  brain injury; global cerebral ischemia; neuroprotection; sodium channels
    DOI:  https://doi.org/10.5114/aoms.2017.72550
  64. Aquat Toxicol. 2019 Feb 27. pii: S0166-445X(18)31134-2. [Epub ahead of print]210 242-250
    Wang H, Zhou L, Liao X, Meng Z, Xiao J, Li F, Zhang S, Cao Z, Lu H.
      Oxine-copper (OxCu) is generally used as an agricultural pesticide and induces harmful effects on ecosystems. In this study, zebrafish was used to assess the aquatic toxicity of OxCu. To detect the effects on development, embryos of 6 h post-fertilization (hpf) were exposed to 10 μg/L, 20 μg/L, 40 μg/L OxCu for 18 h; meanwhile, to evaluate the effects on the behavior, larval fish at 6 days post-fertilization (dpf) were exposed to the same concentrations for 24 h. Here, we show that there are embryonic developmental defects, including abnormalities of head and trunk, brain ventricle atrophy, reduced newborn neurons, disordered neurons, increased intercellular space, concentrated cytoplasm, decreased heart beat and blood flow velocity, and developmental delay of the vascular system; in addition, some embryos exposed to the high concentration of OxCu degraded from the tail. We also found that the spontaneous tail coiling frequency and AChE enzyme activity were reduced, while oxidative stress (free radical damage) and cell apoptosis were significantly increased. Moreover, the expression of genes involved in neurodevelopment, vascular development and apoptosis were dysregulated in the OxCu exposed embryos in a concentration-dependent manner. Finally, we found that after exposure to OxCu, larval locomotor activity was decreased and accompanied by Parkinson-like (increased absolute turn angle and sinuosity) and anxiety-like (preferred to the central area) behavior. These results indicate that OxCu induces developmental toxicity and behavioral alterations by affecting AChE enzyme activity and oxidative stress. Our data present new proofs of OxCu toxicity and a warning for its application.
    Keywords:  Developmental defect; Oxidative stress; Oxine-copper; Zebrafish
    DOI:  https://doi.org/10.1016/j.aquatox.2019.02.020
  65. Oncol Rep. 2019 Mar 13.
    Yang Y, Yang X, Li L, Yang G, Ouyang X, Xiang J, Zhang T, Min X.
      LAG1 longevity assurance homolog 2 (LASS2) is a candidate biomarker in cancer that is dysregulated in various types of tumor, potentially affecting cell growth, invasion and migration. Although its effects on liver cancer metastasis and invasion have been reported, specific phenotypic studies and potential molecular mechanisms have not been completely elucidated in hepatoblastoma (HB). In the present study, the effect of LASS2 on the proliferation, apoptosis and cell cycle of HepG2 HB cells was assessed, and the underlying mechanisms were investigated. The human LASS2 coding sequence was inserted into an adenovirus vector and transduced into HepG2 cells. It was determined that the overexpression of LASS2 inhibited HepG2 cell viability and proliferation, as determined by cell counting kit‑8 and colony formation assays, and induced apoptosis by increasing reactive oxygen species, reducing mitochondrial membrane potential and inducing intracellular Ca2+ overload. In addition, the overexpression of LASS2 induced G0/G1 cell cycle arrest through modulating the expression of cell cycle regulatory proteins, including p27, cyclin D1 and cyclin‑dependent kinase 4. Immunofluorescence was used to determine that nuclear factor (NF)‑κB p‑p65 was primarily expressed in the cytoplasm rather than in the nucleus; western blot analysis demonstrated that LASS2 downregulated the expression of NF‑κB p‑p65 relative to its inactive form in HepG2 cells. These findings suggest that LASS2 inhibits proliferation and induces apoptosis in HepG2 HB cells through the mitochondrial apoptotic, NF‑κB and cell cycle signaling pathways.
    DOI:  https://doi.org/10.3892/or.2019.7058
  66. Atherosclerosis. 2019 Feb 25. pii: S0021-9150(19)30088-7. [Epub ahead of print]284 110-120
    Zhu Y, Zhang Y, Huang X, Xie Y, Qu Y, Long H, Gu N, Jiang W.
      BACKGROUND AND AIMS: Oxidative stress-induced endothelial dysfunction is considered to exert a vital role in the development of atherosclerotic coronary heart disease (CHD). NRF2 is a key transcriptional factor against oxidative stress through activation of multiple ARE-mediated genes. Z-Lig is derived from the Ligusticum species with antitumor, anti-inflammation and neuroprotection activities. However, the antioxidant potentials of Z-Lig on endothelial dysfunction and atherosclerosis have not been well elucidated. Therefore, in the present work, we appraise the cytoprotective property and anti-atherosclerosis effect of Z-Lig.METHODS: Potential NRF2 activators were screened and verified by luciferase reporter gene assay. The protein and mRNA levels of NRF2 and ARE-mediated genes, and GSH/GSSG level in EA.hy926 cells treated with Z-Lig were detected. The cytoprotective property of Z-Lig was assessed in the tert-butyl hydroperoxide (t-BHP)-evoked oxidative stress model. Cell viability and reactive oxygen species (ROS) levels in EA.hy926 cells were determined. An atherosclerosis model induced by HFD was used to determine the anti-atherosclerosis effect of Z-Lig in HFD-fed Ldlr-deficient mice.
    RESULTS: In vitro, 100 μM Z-Lig upregulated expressions of NRF2 and ARE-driven genes, promoted accumulation of nuclear NRF2 and unbound NRF2- KEAP1 complex in EA.hy926 cells. Furthermore, Z-Lig alleviated oxidative stress and cell injury caused by t-BHP via stimulation of the NRF2/ARE pathway. In vivo, intervention with 20 mg/kg Z-Lig markedly restrained atherosclerosis progression, including attenuation of HFD-induced atherosclerotic plaque formation, alleviation of lipid peroxidation and increase in antioxidant enzyme activity in aortas of HFD-fed Ldlr-/- mice. The chemopreventive effects of Z-Lig might be associated with the activation of NRF2 and ARE-driven genes.
    CONCLUSIONS: The present study suggested that Z-Lig is an effective NRF2 activator, which can protect vascular endothelial cells from oxidative stress and rescue HFD-induced atherosclerosis.
    Keywords:  Atherosclerosis; Nrf2; Oxidative stress; Vascular endothelium; Z-Ligustilide
    DOI:  https://doi.org/10.1016/j.atherosclerosis.2019.02.010
  67. Onco Targets Ther. 2019 ;12 1765-1779
    Hou Y, Lan C, Kong Y, Zhu C, Peng W, Huang Z, Zhang C.
      Background and objective: Transcriptional coactivator with PDZ-binding motif (TAZ) has been found to be associated with tumor progression. Mitochondrial homeostasis regulates cancer cell viability and metastasis. However, the roles of TAZ and mitochondrial homeostasis in liver cancer viability have not been explored. The aim of our study was to investigate the influence of TAZ on HepG2 liver cancer cell apoptosis.Materials and methods: HepG2 liver cancer cell was used in the present study, and shRNA against TAZ was transfected into HepG2 cell to knockdown TAZ expression. Mitochondrial function was analyzed using Western blotting, immunofluorescence assay, and flow cytometry. Pathway blocker was used to confirm the role of CaMKII pathway in TAZ-mediated cancer cell death.
    Results: Our results indicated that TAZ deletion induced death in HepG2 cell via apoptosis. Biological analysis demonstrated that mitochondrial stress, including mitochondrial bioenergetics disorder, mitochondrial oxidative stress, and mitochondrial apoptosis, were activated by TAZ deletion. Furthermore, we found that TAZ affected mitochondrial stress by triggering mitochondrial elongation factor 1 (MIEF1)-related mitochondrial dysfunction. The loss of MIEF1 sustained mitochondrial function and promoted cancer cell survival. Molecular investigation illustrated that TAZ regulated MIEF1 expression via the CaMKII signaling pathway. Blockade of the CaMKII pathway prevented TAZ-mediated MIEF1 upregulation and improved cancer cell survival.
    Conclusion: Taken together, our results highlight the key role of TAZ as a master regulator of HepG2 liver cancer cell viability via the modulation of MIEF1-related mitochondrial stress and the CaMKII signaling pathway. These findings define TAZ and MIEF1-related mitochondrial dysfunction as tumor suppressors that act by promoting cancer apoptosis via the CaMKII signaling pathway, with potential implications for new approaches to liver cancer therapy.
    Keywords:  CaMKII signaling pathway; MIEF1; TAZ; death; liver cancer
    DOI:  https://doi.org/10.2147/OTT.S196142
  68. J Integr Med. 2019 Jan 31. pii: S2095-4964(19)30011-1. [Epub ahead of print]
    Wang FY, Jia J, Song HH, Jia CM, Chen CB, Ma J.
      OBJECTIVE: To investigate the protective effect and underlying mechanism(s) of icariin (ICA) in preventing hydrogen peroxide (H2O2)-induced vascular endothelial cell injury via endoplasmic reticulum stress (ERS).METHODS: To study the effects of ICA on H2O2-induced damage, we used the cell counting kit-8 assay to detect cell viability and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay to determine cell adhesion and apoptosis, respectively. Spectrophotometry and enzyme-linked immunosorbent assay were used to measure the expression levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Subsequently, glucose-regulated protein 78 (GRP78), activating transcription factor-4 (ATF4) and eukaryotic initiation factor-2α (eIF2α) were detected using Western blotting.
    RESULTS: In human umbilical vein endothelial cells, different concentrations of ICA exhibited multiple effects, including reduced H2O2 damage, improved cell viability and adhesion, reduced cell apoptosis and increased SOD and GSH-Px activity. Among the ICA concentrations used, only the H2O2 + 100 μmol/L ICA group had significant differences compared to the H2O2 group. ERS activators H2O2 and dl-dithiothreitol (DTT) significantly increased GRP78, ATF4 and eIF2α expressions, decreased cell activity and reduced SOD and GSH-Px activity. In contrast, the H2O2 + 100 μmol/L ICA and H2O2 + 100 μmol/L ICA + DTT groups had significant inhibitory effects on the expressions of GRP78, ATF4 and eIF2α proteins, showing enhanced cell viability and SOD and GSH-Px activity.
    CONCLUSION: The results showed the dose-dependent effects of ICA against H2O2-induced injury in vascular endothelial cells. The inhibition of GRP78, ATF4 and eIF2α protein expressions in the ERS, and the subsequent alleviation of oxidative stress damage, might be the molecular mechanism.
    Keywords:  Endoplasmic reticulum stress; Human umbilical vein endothelial cells; Icariin; Oxidative stress
    DOI:  https://doi.org/10.1016/j.joim.2019.01.011
  69. Probiotics Antimicrob Proteins. 2019 Mar 20.
    Wu Y, Xu H, Cao X, Liu R, Tang L, Zeng Z, Li W.
      Probiotics have always been considered as a supplementary therapy for many diseases especially gut disorders. The absorption and barrier function of the gut play a vital role in the maintenance of body homeostasis. This study was to investigate the protective effects of Bacillus amyloliquefaciens SC06 (Ba) on H2O2-induced oxidative stress on intestinal porcine epithelial cells (IPEC-1) based on the level of gene expression. We demonstrated that Ba was a safe probiotic strain in the first place. Results showed that treatment with H2O2 significantly increased the mRNA expression of absorptive transporters glucose transporter 2 (GLUT2), Ala/Ser/Cys/Thr transporter 1 (ASCT1), and ASCT2 compared with the control group. Meanwhile, oxidative stress induced a significant improvement in the mRNA expression of occludin (OCLN) and caspase-3, and remarkably inhibited the expression of L-type amino acid transporter 1 (LAT1) or B cell lymphoma-2 (Bcl-2), respectively. Pretreatment with Ba dramatically reversed the disturbance induced by oxidative stress on the mRNA expression of ASCT1, ASCT2, and OCLN, which also significantly prevented H2O2-inhibited LAT1 and Bcl-2 mRNA expression. However, Ba failed to exert any significant protective effect on GLUT2 and caspase-3 mRNA expression. We concluded that pretreatment with Ba could alleviate the damage caused by oxidative stress to a certain extent and conferred a protective effect to the intestine.
    Keywords:  Ba; H2O2; Oxidative stress; mRNA expression
    DOI:  https://doi.org/10.1007/s12602-019-09538-5
  70. Cell Biosci. 2019 ;9 18
    Zou W, Niu C, Fu Z, Gong C.
      Dexamethasone (Dex) are widely used for the treatment of asthma. However, they may cause apoptosis of bronchial epithelial cells and delay the recovery of asthma. Therefore, it is an urgent problem to find effective drugs to reduce this side effects. Panax notoginseng saponins R1 (PNS-R1) is known to exhibit anti-oxidative and anti-apoptotic properties in many diseases. We aim to investigate whether PNS-R1 can reduce Dex-induced apoptosis in bronchial epithelial cells. In this study, the anti-apoptotic effects of PNS-R1 were investigated by conducting in vitro and in vivo. Annexin V-FITC/PI staining flow cytometry analysis and TUNEL assay were conducted to detect apoptotic cells. Mitochondrial membrane potential was detected by JC-1 analysis. Western blotting and immunohistochemical analysis were conducted to measure caspase3, Bcl-2, Bax, Cyt-c, Apaf-1, cleaved-caspase3 and cleaved-caspase9 levels in lung tissues and 16HBE cells. Our findings demonstrated that Dex could induce apoptosis of bronchial epithelial cells and upregulate caspase3 expression of lung tissues. Western blot showed that Dex increased Bax, Cyt-c, Apaf-1, cleaved-caspase9, cleaved-caspase3 expression and decreased Bcl-2 expression. PNS-R1 could suppress Dex-induced apoptosis of bronchial epithelial cells by inhibiting Bax, Cyt-c, Apaf-1, cleaved-caspase9, cleaved-caspase3 expression and upregulating Bcl-2 expression. Flow cytometry analysis showed PNS-R1 alleviated JC-1 positive cells induced by Dex in 16HBE cells. These results showed that PNS-R1 alleviated Dex-induced apoptosis in bronchial epithelial cells by inhibition of mitochondrial apoptosis pathway. Furthermore, our findings highlighted the potential use of PNS-R1 as an adjuvant drug to treat asthma.
    Keywords:  Apoptosis; Asthma; Bronchial epithelial cells; Dex; PNS-R1
    DOI:  https://doi.org/10.1186/s13578-019-0279-x
  71. Stroke. 2019 Mar 20. STROKEAHA118021590
    Zhang T, Wu P, Budbazar E, Zhu Q, Sun C, Mo J, Peng J, Gospodarev V, Tang J, Shi H, Zhang JH.
      Background and Purpose- Mitoquinone has been reported as a mitochondria-targeting antioxidant to promote mitophagy in various chronic diseases. Here, our aim was to study the role of mitoquinone in mitophagy activation and oxidative stress-induced neuronal death reduction after subarachnoid hemorrhage (SAH) in rats. Methods- Endovascular perforation was used for SAH model of male Sprague-Dawley rats. Exogenous mitoquinone was injected intraperitoneally 1 hour after SAH. ML385, an inhibitor of Nrf2 (nuclear factor-E2-related factor 2), was given intracerebroventricularly 24 hours before SAH. Small interfering RNA for PHB2 (prohibitin 2) was injected intracerebroventricularly 48 hours before SAH. Nuclear, mitochondrial, and cytoplasmic fractions were gathered using nucleus and mitochondria isolation kits. SAH grade evaluation, short- and long- term neurological function tests, oxidative stress, and apoptosis measurements were performed. Pathway related proteins were investigated with Western blot and immunofluorescence staining. Results- Expression of Keap1 (Kelch-like epichlorohydrin-associated protein 1, 2.84× at 24 hours), Nrf2 (2.78× at 3 hours), and LC3II (light chain 3-II; 1.94× at 24 hours) increased, whereas PHB2 (0.46× at 24 hours) decreased after SAH compared with sham group. Mitoquinone treatment attenuated oxidative stress and neuronal death, both short-term and long-term. Administration of mitoquinone resulted in a decrease in expression of Keap1 (0.33×), Romo1 (reactive oxygen species modulator 1; 0.24×), Bax (B-cell lymphoma-2 associated X protein; 0.31×), Cleaved Caspase-3 (0.29×) and an increase in Nrf2 (2.13×), Bcl-xl (B-cell lymphoma-extra large; 1.67×), PINK1 (phosphatase and tensin-induced kinase 1; 1.67×), Parkin (1.49×), PHB2 (1.60×), and LC3II (1.67×) proteins compared with SAH+vehicle group. ML385 abolished the treatment effects of mitoquinone on behavior and protein levels. PHB2 small interfering RNA reversed the outcomes of mitoquinone administration through reduction in protein expressions downstream of PHB2. Conclusions- Mitoquinone inhibited oxidative stress-related neuronal death by activating mitophagy via Keap1/Nrf2/PHB2 pathway after SAH. Mitoquinone may serve as a potential treatment to relieve brain injury after SAH.
    Keywords:  Kelch-like epichlorohydrin-associated protein 1; mitoquinone; nuclear factor E2-related factor 2; prohibitin 2; rats; subarachnoid hemorrhage
    DOI:  https://doi.org/10.1161/STROKEAHA.118.021590
  72. Mol Med Rep. 2019 Mar 19.
    Shao D, Wu Z, Bai S, Fu G, Zou Z.
      The present study aimed to investigate the function of micro (mi)RNA‑153 against isoflurane‑induced neurotoxicity and its mechanism. In isoflurane‑induced mice, miRNA‑153 expression was downregulated compared with in the control group. Downregulation of miRNA‑153 induced neurocyte apoptosis, reduced cell growth and promoted oxidative stress in an in vitro model. Overexpression of miRNA‑153 reduced oxidative stress, promoted cell growth and inhibited neurocyte apoptosis within an in vitro model. Downregulation of miRNA‑153 suppressed nuclear erythroid‑2 related factor 2 (Nrf2)/antioxidant response element (ARE) signaling pathway, which was induced via the overexpression of miRNA‑153 in vitro. The Nrf2 agonist, dimethyl fumarate (2.5 µM), induced the Nrf2/ARE signaling pathway and reduced oxidative stress to induce neurocyte apoptosis in vitro following treatment with anti‑miRNA‑153. The results of the present study suggested the function of miRNA‑153 against neurotoxicity via Nrf2/ARE‑mediated cytoprotection.
    DOI:  https://doi.org/10.3892/mmr.2019.10056
  73. Curr Res Transl Med. 2019 Mar 19. pii: S2452-3186(19)30007-8. [Epub ahead of print]
    Wang J, De-Qiong X, Hong DQ, Zhang QQ, Zhang J.
      Studies have shown that the NRF-2 /HO-1 pathway participates in myocardial ischemic reperfusion injury (MI/R) and that Geniposide (GEN) could protect the myocardial against MI/R. This study aims to examine the protective effects of GEN on MI/R in diabetic rats and further explore the possible mechanism of action. During MI/R in rats, NRF-2 /HO-1signals changed significantly including NRF-2 and HO-1up-regulation, resulting in heart dysfuction, histological damage and increasing oxidative stress and cell apoptosis. Treatment with GEN can significantly improve the general condition and heart function in diabetic rats with decreasing the expression of cTnI, CK-MB, blood glucose, MDA, ROS, cell apoptosis and pathological damage in MI/R. In addition, GEN precondition can also significantly increase the weight of rats and the activity of SOD, CAT and GPx with up-regulating the expression of NRF-2 and HO-1 in MI/R. This study implied that Geniposide has a protective effect on myocardial ischemia reperfusion injury in diabetic rats, and its mechanism is associated with activating NRF2/HO-1 signaling pathway to suppress oxidative stress.
    Keywords:  Diabetes; Geniposide; Myocardial ischemia reperfusion injury; Oxidative stress
    DOI:  https://doi.org/10.1016/j.retram.2019.03.002
  74. Structure. 2019 Mar 04. pii: S0969-2126(19)30051-6. [Epub ahead of print]
    Li M, Mandal A, Tyurin VA, DeLucia M, Ahn J, Kagan VE, van der Wel PCA.
      The peroxidation of cardiolipins by reactive oxygen species, which is regulated and enhanced by cytochrome c (cyt c), is a critical signaling event in mitochondrial apoptosis. We probe the molecular underpinnings of this mitochondrial death signal through structural and functional studies of horse heart cyt c binding to mixed-lipid membranes containing cardiolipin with mono- and polyunsaturated acyl chains. Lipidomics reveal the selective oxidation of polyunsaturated fatty acid (PUFA) cardiolipin (CL), while multidimensional solid-state NMR probes the structure and dynamics of the membrane and the peripherally bound protein. The hydrophilic milieu at the membrane interface stabilizes a native-like fold, but also leads to localized flexibility at the membrane-interacting protein face. PUFA CL acts as both a preferred substrate and a dynamic regulator by affecting the dynamics of the cyt c N70-I85 Ω loop, which covers the heme cavity.
    Keywords:  PUFA; apoptosis; cardiolipin; cytochrome c; lipidomics; membrane oxidation; membrane protein; mitochondrial protein; protein structure and dynamics; solid-state NMR
    DOI:  https://doi.org/10.1016/j.str.2019.02.007
  75. Biochem Pharmacol. 2019 Mar 15. pii: S0006-2952(19)30111-X. [Epub ahead of print]
    Rajesh Y, Biswas A, Kumar U, Banik P, Bharti R, Nayak S, Ghosh SK, Mandal M.
      Glioblastoma (GBM) is the most malignant form of brain tumor posing a major threat to cancer amelioration. Temozolomide (TMZ) resistance is one of the major hurdles towards GBM prognosis. Oxidative stress and ECM remodeling are the two important processes involved in gaining chemo-resistance. Here, we established NFE2L2, an important member of oxidative stress regulation elevated in resistant cells, to be playing a transcriptional regulatory role on MMP-2, an ECM remodeling marker. This link led us to further explore targeted molecules to inhibit NFE2L2, thus affecting MMP-2, an important member promoting chemo-resistance. Thus, diosgenin was proposed as a novel NFE2L2 inhibitor acting as an alternative strategy to prevent the high dose administration of TMZ. Combinatorial therapy of diosgenin and TMZ significantly reduced the dosage regimen of TMZ and also showed affectivity in hitherto TMZ resistant GBM cells. GBM cells underwent apoptosis and early cell cycle arrest with significant reduction in MMP-2 levels. Thus preclinical validation of molecular interaction between diosgenin and NFE2L2 down-regulating MMP-2, EMT markers and promoting apoptosis, offers rationale for new therapeutic horizons in the field of glioblastoma management.
    Keywords:  Diosgenin; Glioblastoma; MMP-2; NFE2L2; Temozolomide resistance
    DOI:  https://doi.org/10.1016/j.bcp.2019.03.025
  76. Drug Chem Toxicol. 2019 Mar 20. 1-6
    Fouad AA, Abdel-Gaber SA, Abdelghany MI.
      The present investigation examined the prospective nephroprotective effect of hesperidin (HSN) in mice challenged with a single i.p. injection of cyclophosphamide (CPE) at a dose of 200 mg/kg. HSN (100 and 200 mg/kg/day, p.o.) was given for 10 days, starting 5 days prior to CPE administration. HSN significantly reduced the CPE-induced increments of serum creatinine and cystatin C. HSN also significantly reduced malondialdehyde, nitric oxide, Bax/Bcl-2 ratio, and caspase-3, and significantly raised total antioxidant capacity, and interleukin-10/tumor necrosis factor-α ratio in kidneys of mice received CPE. In addition, HSN significantly prevented the histopathological injury, and kidney injury molecule-1 expression in kidneys of mice given CPE. It was concluded that HSN guarded against nephrotoxic effect of CPE in mice by tackling oxidative/nitrative stress, inflammation, and apoptosis.
    Keywords:  Hesperidin; apoptosis; cyclophosphamide; inflammation; kidney; mice; oxidative stress
    DOI:  https://doi.org/10.1080/01480545.2018.1560467