摘要
目的探讨淫羊藿次苷Ⅱ(IcarisideⅡ,ICSⅡ)对过氧化氢(H_(2)O_(2))诱导的心肌细胞氧化应激损伤的影响与机制。方法培养人心肌细胞(AC16),筛选ICSⅡ、NADPH氧化酶(NOXs)抑制剂(Apocynin)和NOXs激活剂(TBCA)的作用浓度。将AC16细胞分为对照组、H_(2)O_(2)组、Apocynin组(20μmol/L)、ICSⅡ组(40μmol/L)、ICSⅡ+TBCA组(40μmol/L ICSⅡ+5μmol/L TBCA),除对照组外其余组采用H_(2)O_(2)(200μmol/L)诱导。CCK-8法检测细胞活力;TUNEL法检测心肌细胞凋亡;DCFH-DA荧光探针检测细胞内活性氧(ROS)水平;试剂盒检测细胞乳酸脱氢酶(LDH)释放量、丙二醛(MDA)水平和超氧化物歧化酶(SOD)活性;Western Blot检测细胞NOX2蛋白表达;qRT-PCR检测细胞NOX2 mRNA水平。结果40μmol/L ICSⅡ、20μmol/L Apocynin和5μmol/L TBCA为最佳作用浓度。与对照组相比,H_(2)O_(2)组细胞活力、SOD活性降低,细胞凋亡率、ROS相对荧光强度、LDH、MDA水平、NOX2 mRNA和蛋白水平升高(P<0.05);与H_(2)O_(2)组相比,Apocynin组、ICSⅡ组细胞活力、SOD活性升高,细胞凋亡率、ROS相对荧光强度、LDH、MDA水平、NOX2 mRNA和蛋白水平降低(P<0.05);TBCA可明显削弱ICSⅡ对心肌细胞的保护作用(P<0.05)。结论ICSⅡ可能通过抑制NOX2表达,减轻H_(2)O_(2)诱导的心肌细胞氧化应激,抑制心肌细胞凋亡。
Objective To investigate the effects and mechanisms of icarisideⅡ(ICSⅡ)on oxidative stress injury induced by hydrogen peroxide(H_(2)O_(2))in cardiomyocytes.Methods Human cardiac muscle cells(AC16)were cultured to determine the optimal working concentrations of ICSⅡ,the NADPH oxidase(NOXs)inhibitor Apocynin,and the NOXs activator TBCA.Cells were grouped into control group,H_(2)O_(2) group,Apocynin group(20μmol/L),ICSⅡgroup(40μmol/L),and ICSⅡ+TBCA group(40μmol/L ICSⅡ+5μmol/L TBCA).Except the control group,all other groups were induced with H_(2)O_(2)(200μmol/L).CCK-8 method was applied to detect cell viability.TUNEL method was applied to detect myocardial cell apoptosis.DCFH-DA fluorescent probe was applied to detect intracellular reactive oxygen species(ROS)level.The reagent kit was used to detect the release of lactate dehydrogenase(LDH),malondialdehyde(MDA)levels,and superoxide dismutase(SOD)activity in cells.Western Blot was applied to detect the expression of NOX2 protein in cells.QRT-PCR was applied to detect the level of NOX2 mRNA in cells.Results The optimal concentrations were 40μmol/L ICSⅡ,20μmol/L Apocynin,and 5μmol/L TBCA.Compared with the control group,the cell viability and SOD activity in the H_(2)O_(2) group decreased,while the apoptosis rate,relative fluorescence intensity of ROS,LDH,MDA levels,NOX2 mRNA and protein levels increased(P<0.05).Compared with the H_(2)O_(2) group,the cell viability and SOD activity increased in the Apocynin group and ICSⅡgroup,while the apoptosis rate,relative fluorescence intensity of ROS,LDH,MDA levels,NOX2 mRNA and protein levels decreased(P<0.05).TBCA could obviously weaken the protective effect of ICSⅡon myocardial cells(P<0.05).Conclusion ICSⅡmay alleviate H_(2)O_(2)-induced oxidative stress and inhibit cardiomyocyte apoptosis by suppressing NOX2 expression.
作者
吴立国
马蓉
闫广有
耿学斌
Wu Liguo;Ma Rong;Yan Guangyou(Department of Cardiology,Nanhu Hospital Tangshan,Tangshan,Hebei 063000;Department of Cardiology,982 Hospital of the Joint Service Support Force of the Chinese People's Liberation Army,Tangshan,Hebei 063000,China)
出处
《四川医学》
2025年第12期1344-1350,共7页
Sichuan Medical Journal
基金
河北省2024年度医学科学研究课题计划(编号:20241923)。
