丹参提取物联合瑞芬太尼通过调控Nrf2信号通路减轻缺氧复氧诱导的心肌细胞损伤

Salvia miltiorrhiza Bge.Extract Combined with Remifentanil Attenuates Hypoxia/Reoxygenation-induced Cardiomyocyte Injury by Regulating Nrf2 Signaling Pathway

为探讨丹参提取物联合瑞芬太尼在减轻缺氧/复氧诱导的心肌细胞损伤中的作用及其机制。本研究将H9c2心肌细胞分为对照组、模型组、丹参提取物组(Tan I组)、瑞芬太尼组(RPC)组和联合组。对照组细胞不做处理,模型组缺氧/复氧(HR)诱导的心肌细胞损伤模型,Tan I组在模型组的基础上给与1μmol/LTan I孵育24h,RPC组在模型组的基础上给与1μmol/LRPC孵育24h联合组在模型组的基础上同时给与1μmol/L Tan I和1μmol/L RPC孵育24 h。采用CCK-8测定各组细胞活力流式细胞术测定各组细胞凋亡率,采用Western blot测定各组细胞Nrf2和HO-1表达水平进一步转染Nrf2-siRNA分析Nrf2信号通路对Tan I联合RPC保护HR诱导H9c2心肌细胞损伤的影响。结果表明,HR诱导后H9c2心肌细胞的活力显著下降,药物处理后,Tan I组、RPC组和联合组H9c2细胞的细胞活力显著增加,且联合组的细胞活力改善与Tan I组和RPC组相比更显著;HR诱导后H9c2心肌细胞的凋亡率显著升高,药物处理后,Tan I组、RPC组和联合组H9c2细胞的细胞凋亡率显著下降,且联合组的细胞凋亡率下降水平与Tan I组和RPC组相比更显著;HR引起H9c2细胞Nrf2总蛋白表达和核Nrf2蛋白表达水平显著降低,各组给药后增加了总Nrf2蛋白和核Nrf2蛋白的表达,降低了细胞质中的Nrf2蛋白,且与Tan I组和RPC组相比联合组总Nrf2蛋白和核Nrf2蛋白的表达水平显著增高,细胞质中的Nrf2蛋白表达水平显著降低;转染Nrf2-siRNA后,Tan I联合RPC处理的H9c2心肌细胞中Nrf2和HO-1的表达均下降。丹参提取物联合瑞芬太尼可通过调控Nrf2信号通路减轻HR诱导的心肌细胞损伤。

In order to investigate the effect and mechanismof Salvia miltiorrhiza Bge.extract combined with remifentanil in alleviating hypoxia/reoxygenation-induced myocardial cell injury.H9c2 cardiomyocytes were divided into control group,model group,danshen extract group(Tan I group),RPC group and combination group.The cells in the control group were not treated.The myocardial cell injury model was induced by hypoxia/reoxygenation(HR)in the model group.The Tan I group was incubated with 1 mol/L Tan I for 24 h on the basis of the model group.The RPC group was incubated with 1 mol/L RPC for 24 h on the basis of the model group.The combined group was incubated with 1 mol/L Tan I and 1 mol/L RPC for 24 h on the basis of the model group.The cell viability of each group was determined by CCK-8,the apoptosis rate of each group was determined by flow cytometry,and the expression levels of Nrf2 and HO-1 in each group were determined by western blot.Nrf2-siRNA was further transfected to analyze the effect of Nrf2 signaling pathway on the protection of Tan I combined with RPC against HR-induced H9c2 cardiomyocyte injury.The viability of H9c2 cardiomyocytes was significantly decreased after HR induction.After drug treatment,the cell viability of H9c2 cells in Tan I group,RPC group and combination group was significantly increased,and the improvement of cell viability in combination group was more significant than that in Tan I group and RPC group.The apoptosis rate of H9c2 cardiomyocytes increased significantly after HRinduction.After drug treatment,the apoptosis rate of H9c2 cells in Tan I group,RPCgroup and combination group decreased significantly,and the decrease of apoptosis rate in combination group was more significant than that in Tan I group and RPC group.HRcaused a significant decrease in the expression of total Nrf2 protein and nuclear Nrf2 protein in H9c2 cells.After administration,the expression of total Nrf2 protein and nuclear Nrf2 protein was increased,and the Nrf2 protein in the cytoplasm was decreased.Compared with Tan I group and RPC group,the expression levels of total Nrf2 protein and nuclear Nrf2 protein in the combined group were significantly increased,and the expression level of Nrf2 protein in the cytoplasm was significantly decreased.After transfection of Nrf2-siRNA,the expression of Nrf2 and HO-1 in H9c2 cardiomyocytes treated with Tan I combined with RPC decreased.Salvia miltiorrhiza extract combined with remifentanil attenuates HR-induced cardiomyocyte injury by modulating the Nrf2 signalling pathway.