Department of Cardiac Surgery, Affiliated Hospital of Jining Medical University, Jining, Shandong, China.
Biomed Res Int. 2020 Jun 24;2020:8590861. doi: 10.1155/2020/8590861. eCollection 2020.
This project is aimed at investigating whether CircANXA2 can promote the apoptosis of myocardial cells by inhibiting miR-133 expression and thereby participate in the development of myocardial ischemia-reperfusion injury. . Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of CircANXA2 in H9c2 cells after hypoxia/reoxygenation (H/R) treatment. Evaluation of myocardial injury markers in H9c2 cells was performed using commercial kits, including lactate dehydrogenase (LDH), malonaldehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidation (GSH-PX). MTT analysis and flow cytometry were used to detect myocardial cell proliferation and apoptosis, respectively. Western blot was used to detect the protein expression of apoptosis-related genes.
qRT-PCR results showed that compared with the control, the expression of CircANXA2 was upregulated and the expression level of miR-133 was significantly decreased in H/R-treated H9c2 cells. CircANXA2 overexpression increased LDH, MDA, SOD, and GSH-PX activity in H/R-treated H9c2 cells. At the same time, CircANXA2 overexpression inhibited the proliferation of H/R-treated cells, and CircANXA2 was able to induce cardiomyocyte apoptosis. Western blot results showed that after overexpression of CircANXA2, the proapoptotic genes Bax and cytochrome C was upregulated, while the antiapoptotic gene Bcl-2 was downregulated. In H9c2 cells, upregulating miR-133 can reverse the inhibition of proliferation induced by CircANXA2 overexpression and increase apoptosis.
CircANXA2 promotes cardiomyocyte apoptosis in myocardial ischemia-reperfusion injury by inhibiting the expression of miR-133. CircANXA2 may be a potential target for myocardial ischemia-reperfusion injury.
本项目旨在研究 CircANXA2 是否可以通过抑制 miR-133 的表达来促进心肌细胞凋亡，从而参与心肌缺血再灌注损伤的发生。采用定量实时聚合酶链反应（qRT-PCR）检测缺氧/复氧（H/R）处理后 H9c2 细胞中 CircANXA2 的表达水平。采用商业试剂盒检测 H9c2 细胞中心肌损伤标志物，包括乳酸脱氢酶（LDH）、丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-PX）。MTT 分析和流式细胞术分别用于检测心肌细胞增殖和凋亡。Western blot 用于检测凋亡相关基因的蛋白表达。
qRT-PCR 结果显示，与对照组相比，H/R 处理的 H9c2 细胞中 CircANXA2 的表达上调，miR-133 的表达水平明显降低。CircANXA2 过表达增加了 H/R 处理的 H9c2 细胞中的 LDH、MDA、SOD 和 GSH-PX 活性。同时，CircANXA2 过表达抑制了 H/R 处理细胞的增殖，并且 CircANXA2 能够诱导心肌细胞凋亡。Western blot 结果显示，过表达 CircANXA2 后，促凋亡基因 Bax 和细胞色素 C 上调，而抗凋亡基因 Bcl-2 下调。在 H9c2 细胞中，上调 miR-133 可以逆转 CircANXA2 过表达引起的增殖抑制并增加凋亡。
CircANXA2 通过抑制 miR-133 的表达促进心肌缺血再灌注损伤中的心肌细胞凋亡。CircANXA2 可能是心肌缺血再灌注损伤的潜在靶点。
