GAPDH-knockdown reduce rotenone-induced H9C2 cells death via autophagy and anti-oxidative stress pathway
GAPDH-knockdown reduce rotenone-induced H9C2 cells death via autophagy and anti-oxidative stress pathway
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GAPDH 敲低通过自噬和抗氧化应激途径减少鱼藤酮诱导的 H9C2 细胞死亡。
DOI：
10.1016/j.toxlet.2015.02.017
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发表时间：
2015-05-05
影响因子：
3.5
通讯作者：
Zhang Ping
中科院分区：
文献类型：
--
作者：
Shao Liang;Figtree, Gemma;Zhang Ping
背景：GAPDH 以其管家功能而闻名，也被证明参与饥饿、化学损伤和氧化应激等应激条件下的细胞损伤、凋亡和死亡。本研究探讨了 GAPDH 敲低对鱼藤酮反应引起的细胞损伤的影响。方法：在暴露于鱼藤酮（0 nM、20 nM、40 nM 和 80 nM）之前，使用 siRNA 敲低 H9C2 成肌细胞中的 GAPDH。通过蛋白质印迹检测自噬蛋白（Beclin-1、Atg5、LC-3A/B 和 p62）和 MDC 染色检测酸性物质。使用促凋亡蛋白和流式细胞术评估细胞凋亡和死亡，并测量细胞内ATP相对浓度。通过测量 DCFH-DA、TBARS、GSH 和 SOD 来评估氧化应激。结果：在这项研究中，GAPDH 敲除增强了鱼藤酮诱导的 H9C2 细胞的自噬，减少了氧化应激并增加了抗氧化途径；并减少细胞凋亡和死亡。此外，GAPDH 敲低保留了细胞能量。结论：siRNA 介导的 GAPDH 敲低通过自噬和抗氧化应激途径减少了鱼藤酮诱导的 H9C2 细胞死亡。这项研究丰富了对GAPDH病理生理学作用的理解，并为以氧化应激为特征的心脏病状态提供了潜在的新治疗靶点。 (C) 2015 Elsevier Ireland Ltd. 保留所有权利。
Background: GAPDH, well known for its house-keeping functions, has also been shown to be involved in cell injury, apoptosis and death under conditions of stress such as starvation, chemical injury and oxidative stress. This study examines the effect of GAPDH knockdown on cell injury in response to Rotenone.Methods: GAPDH was knocked down in H9C2 cardiomyoblasts using siRNA prior to exposure to rotenone (0 nM, 20 nM, 40nM and 80 nM). Autophagy was detected by western blot for autophagy proteins (Beclin-1, Atg5, LC-3A/B and p62) and MDC staining for acidic substances. Pro-apoptosis protein and flow cytometry were used to assess cell apoptosis and death and intracellular ATP relative concentration was measured. Oxidant stress was assessed by measuring DCFH-DA, TBARS, GSH and SOD.Results: In this study, GAPDH-knockdown enhanced autophagy in rotenone-induced H9C2 cells, decreased oxidant stress and increased antioxidant pathways; and reduced cell apoptosis and death. Furthermore, GAPDH-knockdown preserved cell energy.Conclusion: siRNA-mediated GAPDH knockdown reduced rotenone-induced H9C2 cell death occurring via autophagy and anti-oxidative stress pathway. This study enriches the understanding of GAPDH pathophysiology role, and provides potential new therapeutic targets for cardiac disease states characterized by oxidative stress. (C) 2015 Elsevier Ireland Ltd. All rights reserved.