外源性NO供体对缺氧/复氧诱导H9c2细胞凋亡、肌钙蛋白及组蛋白去乙酰化酶的影响  

Effects of Exogenous NO Donor on Apoptosis,Troponin,and Histone Deacetylase of H9c2 Cells Induced by Hypoxia/Reoxygenation

作　　者：李俐 王志鑫 吕雅萱 郭彦青 LI Li;WANG Zhixin;LYU Yaxuan;GUO Yanqing(Shanxi Cardiovascular Hospital,Taiyuan 030024,Shanxi,China)

机构地区：[1]山西省心血管病医院,太原030024

出　　处：《中西医结合心脑血管病杂志》2025年第7期1022-1027,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：山西省心血管病医院科研激励计划项目(No.XYS20200108,XYS20220101);山西省卫生健康委员会人才专项科研项目(No.2020036)。

摘　　要：目的:探讨外源性一氧化氮(NO)供体对缺氧/复氧诱导H9c2细胞损伤的影响。方法:将H9c2细胞分为对照组、缺氧/复氧组和缺氧/复氧+NO组[外源性NO供体硝普钠(SNP)处理H9c2细胞]。通过细胞计数试剂盒(CCK-8)检测细胞活力;流式细胞仪检测细胞凋亡率;免疫荧光法检测各组心肌肌钙蛋白T(cTnT)表达;蛋白免疫印迹法(Western Blot)检测慢肌肌钙蛋白C(TNNC1)、骨骼肌快肌肌钙蛋白I(TNNI2)、肌钙蛋白T(TNNT2)表达;酶联免疫吸附试验(ELISA)试剂盒检测组蛋白去乙酰化酶(HDACs)水平。结果:1 mmol/L浓度的SNP溶液处理H9c2细胞48 h,细胞活性最高。与对照组相比,缺氧/复氧组细胞凋亡率明显升高(P<0.01),而缺氧/复氧+NO组细胞凋亡率明显降低(P<0.05),缺氧/复氧+NO组细胞凋亡率明显低于缺氧/复氧组(P<0.01)。与对照组相比,缺氧/复氧组TNNC1、TNNI2和TNNT2蛋白表达水平明显升高(P<0.05);与缺氧/复氧组比较,TNNI2和TNNT2蛋白表达水平下降(P<0.05);缺氧/复氧组HDACs浓度低于对照组,缺氧/复氧+NO组HDACs浓度高于缺氧/复氧组,差异均有统计学意义(P<0.01)。结论:外源性NO供体可能通过抑制心肌细胞凋亡、降低肌钙蛋白表达、上调HDACs等多种途径保护H9c2心肌细胞免受缺氧/复氧损伤。ObjectiveObjective:To explor the effect of exogenous nitric oxide(NO)donor on hypoxia/reoxygenation induced H9c2 cell damage.MethodsMethods:H9c2 cells were divided into control group,hypoxia/reoxygenation group,and hypoxia/reoxygenation+NO group[H9c2 cells treated with exogenous NO donor sodium nitroprusside(SNP)].Cell viability was detected by cell counting reagent(CCK-8).The apoptosis rate was detected by flow cytometry.The expression of cardiac troponin T(cTnT)was detected by immunofluorescence.The expression of troponin C(TNNC1)in slow muscle,troponin I(TNNI2)in fast muscle,and troponin T(TNNT2)in skeletal muscle were detected by Western Blot.The level of histone deacetylases(HDACs)was detected by enzyme-linked immunosorbent assay(ELISA)kit.ResultsResults:H9c2 cells were treated with 1 mmol/L SNP solution for 48 h,and the cell activity was the highest.Compared with the control group,the apoptosis rate of hypoxia/reoxygenation group increased(P<0.01),while the apoptosis rate of hypoxia/reoxygenation+NO group decreased(P<0.05),and the apoptosis rate of hypoxia/reoxygenation+NO group lower than that of the hypoxia/reoxygenation group(P<0.01).Compared with the control group,the expression levels of TNNC1,TNNI2,and TNNT2 protein in hypoxia/reoxygenation group increased(P<0.05),the expression levels of TNNI2 and TNNT2 protein decreased compared with those of the hypoxia/reoxygenation group(P<0.01).The HDACs concentration in hypoxia/reoxygenation group was lower than that in the control group,and the HDACs concentration in hypoxia/reoxygenation+NO group was higher than that in the hypoxia/reoxygenation group,the differences were statistically significant(P<0.01).ConclusionConclusion:Exogenous NO donors may protect H9c2 cardiomyocytes from hypoxia/reoxygenation injury by inhibiting myocardial apoptosis,reducing troponin expression,and upregating HDACs.

关 键 词：外源性一氧化氮 缺氧/复氧 H9C2细胞 缺血再灌注损伤 硝普钠 实验研究 

分 类 号：R541[医药卫生—心血管疾病]