Zhao Yintao, Pei Xiaoxin, Liu Yuan, Xu Yawei, Peng Meng, Yang Haibo
Department of Cardiovascular Medicine, the First Affiliated Hospital of Zhengzhou University, Henan 450052, China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2021 Dec 10;38(12):1199-1203. doi: 10.3760/cma.j.cn511374-20200915-00669.
To study the effect of down-regulating miR-488 targeting Jag1 on the injury of hypoxia-reoxygenation myocardial H9c2 cells.
A hypoxic-reoxygenated myocardial H9c2 cell injury model was constructed. miR-488 inhibitor was used to transfect the cells. CCK-8 method and flow cytometry were used to detect cell proliferation and apoptosis in each group. Lactate dehydrogenase (LDH), superoxide dismutase (SOD), malonaldehyde (MDA), catalase (CAT) levels were detected. Western blotting was used to detect the expression of Bcl-2 associated X Protein (Bax) and B cell lymphoma/lewkmia-2 (Bcl-2). Target genes of miR-488 were predicted, and a luciferase reporter system was used to verify the targeting relationship between the two. Myocardial H9c2 cells were co-transfected with miR-488 inhibitor and Jag1 siRNA, and treated with hypoxia and reoxygenation, cell proliferation, apoptosis, LDH, SOD, MDA, CAT levels, and Bax, Bcl-2 protein expression were detected.
The expression of miR-488 in the hypoxia-reoxygenated myocardial H9c2 cells was increased, along with reduced cell proliferation, increased apoptosis, increased Bax protein expression, decreased Bcl-2 protein expression, increased MDA, decreased CAT and SOD, and increased LDH level in the supernatant of cell culture. When myocardial H9c2 cells were transfected with miR-488 inhibitor and treated with hypoxia and reoxygenation, the expression of miR-488 was decreased, along with increased cell proliferation, decreased apoptosis, decreased Bax protein expression, increased Bcl-2 protein expression, decreased MDA, increased CAT and SOD, and decreased LDH level in the supernatant of cell culture. Down-regulation of miR-488 could target and down-regulate Jag1 expression. And Jag1 siRNA could reverse the effect of miR-488 inhibitor on the proliferation, apoptosis, LDH, SOD, MDA, CAT levels and the expression of Bax and Bcl-2 of hypoxic-reoxygenated myocardial H9c2 cells.
Down-regulating miR-488 targeted Jag1 can attenuate hypoxia-reoxygenation induced myocardial H9c2 cell injury.
研究下调靶向Jag1的miR-488对缺氧复氧心肌H9c2细胞损伤的影响。
构建缺氧复氧心肌H9c2细胞损伤模型。用miR-488抑制剂转染细胞。采用CCK-8法和流式细胞术检测各组细胞增殖和凋亡情况。检测乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）、丙二醛（MDA）、过氧化氢酶（CAT）水平。采用蛋白质免疫印迹法检测Bcl-2相关X蛋白（Bax）和B细胞淋巴瘤/白血病-2（Bcl-2）的表达。预测miR-488的靶基因，并用荧光素酶报告系统验证二者的靶向关系。将miR-488抑制剂与Jag1 siRNA共转染心肌H9c2细胞，进行缺氧复氧处理，检测细胞增殖、凋亡、LDH、SOD、MDA、CAT水平以及Bax、Bcl-2蛋白表达。
缺氧复氧心肌H9c2细胞中miR-488表达升高，同时细胞增殖减少、凋亡增加、Bax蛋白表达增加、Bcl-2蛋白表达减少、MDA增加、CAT和SOD减少，细胞培养上清液中LDH水平升高。用miR-488抑制剂转染心肌H9c2细胞并进行缺氧复氧处理后，miR-488表达降低，同时细胞增殖增加、凋亡减少、Bax蛋白表达减少、Bcl-2蛋白表达增加、MDA减少、CAT和SOD增加，细胞培养上清液中LDH水平降低。下调miR-488可靶向并下调Jag1表达。Jag1 siRNA可逆转miR-488抑制剂对缺氧复氧心肌H9c2细胞增殖、凋亡、LDH、SOD、MDA、CAT水平以及Bax和Bcl-2表达的影响。
下调靶向Jag1的miR-488可减轻缺氧复氧诱导的心肌H9c2细胞损伤。
