金樱子总黄酮上调miR-1247-3p表达对缺氧/复氧诱导心肌细胞损伤的影响  

The Effect of Rosa Laevigata Michx Total Flavonoids on Hypoxia/Reoxygenation-Induced Cardiomyocyte Injury by Up-Regulating the Expression of miR-1247-3p

作　　者：刘晓东 刘晓琴 闫业军[1] LIU Xiaodong;LIU Xiaoqin;YAN Yejun(Baoji Central Hospital,Baoji 721008,Shaanxi,China)

机构地区：[1]宝鸡市中心医院,陕西宝鸡721008

出　　处：《中西医结合心脑血管病杂志》2023年第4期649-654,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：陕西省自然科学基础研究计划项目(No.2021JM-587)。

摘　　要：目的:观察金樱子总黄酮(RLMTF)对缺氧/复氧(H/R)诱导的心肌细胞损伤的影响及可能作用机制。方法:采用H/R诱导大鼠心肌细胞H9C2建立细胞损伤模型,采用不同剂量的金樱子总黄酮处理细胞,实验分为Con组、H/R组、H/R+RLMTF-L组、H/R+RLMTF-M组、H/R+RLMTF-H组、H/R+miR-NC组、H/R+miR-1247-3p组、H/R+RLMTF+anti-miR-NC组、H/R+RLMTF+anti-miR-1247-3p组。使用试剂盒检测丙二醛(MDA)水平与超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性;流式细胞术检测细胞凋亡率;实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-1247-3p表达。结果:与Con组比较,H/R组MDA水平、细胞凋亡率和Cleaved-Caspase 3、Cleaved-Caspase 9蛋白水平升高(P<0.05),miR-1247-3p表达量和SOD、GSH-Px活性降低(P<0.05)。与H/R组比较,H/R+RLMTF-L组、H/R+RLMTF-M组、H/R+RLMTF-H组MDA水平、细胞凋亡率降低(P<0.05),miR-1247-3p表达量和SOD、GSH-Px活性升高(P<0.05),且呈剂量依赖性。与H/R+miR-NC组比较,H/R+miR-1247-3p组MDA水平、细胞凋亡率降低(P<0.05),SOD、GSH-Px活性升高(P<0.05);与H/R+RLMTF+anti-miR-NC组比较,H/R+RLMTF+anti-miR-1247-3p组MDA水平、细胞凋亡率升高(P<0.05),SOD、GSH-Px活性降低(P<0.05)。结论:金樱子总黄酮通过促进miR-1247-3p表达,抑制氧化应激及细胞凋亡,从而减轻H/R诱导的心肌细胞损伤。Objective: To explore the effect of rosa laevigata michx total flavonoids(RLMTF)on the cardiomyocyte injury induced by hypoxia/reoxygenation(H/R).Methods: H/R-induced rat cardiomyocyte H9C2 was used to establish a cell injury model, and different doses of rosa laevigata michx total flavonoids were used to treat the cells.Experimental groups were Con group, H/R group, H/R+RLMTF-L group, H/R+RLMTF-M group, H/R+RLMTF-H group, H/R+miR-NC group, H/R+miR-1247-3p group, H/R+RLMTF+anti-miR-NC group, H/R+RLMTF+anti-miR-1247-3p group.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-Px) activities were measured using kits.The apoptosis rate was measured by flow cytometry.The expression of miR-1247-3p was measured by real-time fluorescence quantitative enzyme chain polymerization reaction(qRT-PCR).Results: Compared with the Con group, the level of MDA,apoptosis rate, the levels of Cleaved-Caspase 3 and Cleaved-Caspase 9 protein in the H/R group were increased(P<0.05),while the expressions of miR-1247-3p, and the activities of SOD,GSH-Px were decreased(P<0.05).Compared with the H/R group, the level of MDA,apoptosis rate, the levels of Cleaved-Caspase 3 and Cleaved-Caspase 9 protein in the H/R+RLMTF-L group, H/R+RLMTF-M group, and H/R+RLMTF-H group were decreased(P<0.05),while the expressions of miR-1247-3p, and the activities of SOD,GSH-Px were increased(P<0.05),with dose-dependence.Compared with the H/R+miR-NC group, the level of MDA,and apoptosis rate in the H/R+miR-1247-3p group were decreased(P<0.05),while the activities of SOD and GSH-Px were increased(P<0.05).Compared with the H/R+RLMTF+anti-miR-NC group, the level of MDA,and apoptosis rate in the H/R+RLMTF+anti-miR-1247-3p group were increased(P<0.05),while the activities of SOD and GSH-Px were decreased(P<0.05).Conclusion: Rosa laevigata michx total flavonoids could inhibit oxidative stress and apoptosis by promoting the expression of miR-1247-3p, thereby reducing H/R-induced cardiomyocyte injury.

关 键 词：心肌细胞 H9C2 金樱子总黄酮 缺氧/复氧 miR-1247-3p 氧化应激 细胞凋亡 大鼠 实验研究 

分 类 号：R285[医药卫生—中药学]