Mechanism of interactions between endoplasmic reticulum stress and autophagy in hypoxia/reoxygenation-induced injury of H9c2 cardiomyocytes
Mechanism of interactions between endoplasmic reticulum stress and autophagy in hypoxia/reoxygenation-induced injury of H9c2 cardiomyocytes
复制标题
H9c2心肌细胞缺氧/复氧损伤中内质网应激与自噬相互作用的机制
DOI：
10.3892/mmr.2019.10228
复制
发表时间：
2019-07-01
影响因子：
3.4
通讯作者：
Gu, Xiang
中科院分区：
文献类型：
--
作者：
Guan, Gaopeng;Yang, Lei;Gu, Xiang
内质网（ER）应激和自噬参与心肌缺血再灌注（I/R）损伤;然而，它们在这种类型的损伤中的作用尚不清楚。本研究探讨了内质网应激和自噬在H9 c2细胞缺氧/复氧（H/R）损伤中的作用及其机制。CCK-8法检测细胞活力。用mCherry-GFP-LC 3-腺病毒转染监测自噬通量。Western blotting检测自噬相关蛋白和内质网应激相关蛋白的表达水平。流式细胞术和免疫印迹法检测细胞凋亡。结果表明，H/R损伤诱导H9 c2细胞自噬，激活内质网应激，促进细胞凋亡。4-苯基丁酸或C/EBP同源蛋白（CHOP）靶向小干扰RNA（siRNA）抑制ER应激可减少H/R损伤期间的自噬并改善细胞凋亡。雷帕霉素激活自噬可减轻内质网应激，改善细胞凋亡。3-甲基腺嘌呤或Beclin 1靶向siRNA抑制自噬可加重ER应激并加剧细胞凋亡，毒胡萝卜素激活ER应激可降低自噬并诱导细胞凋亡。总的来说，本研究的结果表明，H/R通过ER应激诱导H9 c2细胞的凋亡和自噬，CHOP可能在ER应激诱导的自噬和凋亡中起重要作用。自噬作为一种适应性反应，可被内质网应激激活，减轻内质网应激诱导的细胞凋亡。
Endoplasmic reticulum (ER) stress and autophagy are involved in myocardial ischemia-reperfusion (I/R) injury; however, their roles in this type of injury remain unclear. The present study investigated the roles of ER stress and autophagy, and their underlying mechanisms, in H9c2 cells during hypoxia/reoxygenation (H/R) injury. Cell viability was detected by CCK-8 assay. The autophagy flux was monitored with mCherry-GFP-LC3-adenovirus transfection. The expression levels of autophagy-related proteins and ER stress-related proteins were measured by western blotting. Apoptosis was detected by flow cytometry and western blotting. The results indicated that autophagy was induced, ER stress was activated and apoptosis was promoted in H9c2 cells during H/R injury. The inhibition of ER stress by 4-phenylbutyrate or C/EBP homologous protein (CHOP)-targeting small interfering RNA (siRNA) decreased autophagy and ameliorated cell apoptosis during H/R injury. Activation of autophagy by rapamycin attenuated ER stress and ameliorated cell apoptosis. Inhibition of autophagy by 3-methyladenine or Beclin1-targeting siRNA aggravated ER stress and exacerbated cell apoptosis, and activation of ER stress by thapsigargin decreased autophagy and induced cell apoptosis. Collectively, the findings of the present study demonstrated that H/R induced apoptosis and autophagy via ER stress in H9c2 cells, and that CHOP may serve an important role in ER stress-induced autophagy and apoptosis. Autophagy, as an adaptive response, was activated by ER stress and alleviated ER stress-induced cell apoptosis during H/R injury.