丹皮酚通过抑制miR-155-5p减轻体外培养心肌细胞受到的缺氧/复氧损伤

Alleviation of hypoxia/reoxygenation injury of cardiomyocytes cultured in vitro with paeonol by inhibiting miR-155-5p

目的:探究丹皮酚对受到缺氧/复氧损伤的H9C2细胞的影响及其作用机制。方法:使用或不使用丹皮酚处理H9C2细胞,并建立体外H9C2细胞缺氧/复氧损伤模型。使用qRT-PCR检测miR-155-5p的相对表达水平,通过MTT和流式细胞术检测H9C2细胞的增殖活性和凋亡水平。使用乳酸盐脱氢酶和肌酸激酶对应的ELISA试剂盒检测H9C2细胞损伤情况。结果:丹皮酚显著增强了H9C2细胞活性,减弱了细胞凋亡水平及乳酸盐脱氢酶和肌酸激酶的释放。丹皮酚显著抑制了H9C2细胞中miR-155-5p的表达。单独抑制miR-155-5p产生的效果与丹皮酚一致。同时,使用丹皮酚处理H9C2细胞并向H9C2细胞中转染miR-155-5p mimics后丹皮酚的效果被过表达的miR-155-5p逆转了。结论:丹皮酚通过下调miR-155-5p水平减弱了缺氧/复氧导致的H9C2细胞损伤。

Objective:To investigate the effect of paeonol on H9 C2 cells damaged by hypoxia/reoxygenation and its underlying mechanism.Methods:H9 C2 cells were pre-treated with paeonol or not,and the hypoxia/reoxygenation injury model of H9 C2 cells in vitro was established.Expression level of miR-155-5 p was measured by qRT-PCR.Proliferation activity and apoptosis level of H9 C2 cells were validated using MTT assay and flow cytometry.ELISA kits were used to detect myocardial injury markers creatine kinase(CK)and lactate dehydrogenase(LDH).Results:Paeonol significantly enhanced the activity of H9 C2 cells and decreased the level of apoptosis and the release of CK and LDH after hypoxia/reoxygenation treatment.Paeonol notably inhibited the expression of miR-155-5 p in H9 C2 cells.It was observed that inhibition of miR-155-5 p alone had similar effects on H9 C2 cells with paeonol.After paeonol pre-treatment and miR-155-5 p mimics transfection at the same time,effects of paeonol on H9 C2 cells were reversed by over-expressed miR-155-5 p.Conclusion:Paeonol can alleviate injuries in H9 C2 cells induced by hypoxia/reoxygenation by down-regulating the level of miR-155-5 p.