花旗松素通过抑制NF-κB信号通路对缺氧复氧诱导H9c2心肌细胞损伤的保护作用

Protective effect of Taxifolin on H9c2 myocardial cell injury induced by hypoxia-reoxygenation by inhibiting NF-κB signaling pathway

目的探讨花旗松素通过抑制核转录因子-κB(NF-κB)信号通路对缺氧复氧诱导H9c2心肌细胞损伤的保护作用。方法将H9c2细胞分为A组、B组、C组、D组、E组,A组于37℃、5%CO2恒温箱中培养30 h;B组缺氧(O2浓度<6%的厌氧培养箱)培养24 h,再复氧(37℃、5%CO2恒温箱)培养6 h;C组加入1μmol/L花旗松素,缺氧培养24 h,复氧培养6 h;D组加入10μmol/L花旗松素,缺氧培养24 h,复氧培养6 h;E组加入100μmol/L花旗松素,缺氧培养24 h,复氧培养6 h。比较各组H9c2细胞形态学变化、存活率、凋亡率、NF-κB蛋白表达量、NF-κB抑制蛋白α(IκB-α)蛋白表达量。结果HE染色显示,A组细胞形态结构正常,B组H9c2细胞形态结构异常,横纹模糊,可见大量点状坏死灶,间质水肿明显,炎性细胞浸润明显;C、D、E组H9c2细胞病理改变较B组显著改善,随着花旗松素剂量增大,H9c2细胞病理改善越明显。B组H9c2细胞存活率、IκB-α蛋白表达量较A组明显下降(P<0.05);C、D、E组H9c2细胞存活率、IκB-α蛋白表达量较B组明显升高(P<0.05),呈剂量依赖性。B组H9c2细胞凋亡率、NF-κB蛋白表达量较A组明显升高(P<0.05);C、D、E组H9c2细胞凋亡率、NF-κB蛋白表达量较B组明显下降(P<0.05),呈剂量依赖性。结论花旗松素通过抑制NF-κB信号通路对缺氧复氧诱导H9c2心肌细胞损伤起到保护作用,并这种作用呈剂量依赖性,这可为心肌缺血再灌注损伤的防治提供新的方向。

Objective To investigate the protective effect of taxifolin on H9c2 myocardial cell injury induced by hypoxia-reoxygenation by inhibiting nuclear factor-kappaB(NF-κB)signaling pathway.Methods H9c2 cells were divided into group A,group B,group C,group D,and group E.Group A was cultured in an incubator at 37℃and 5%CO2 for 30 h.Group B was cultured for 24 hours under hypoxia(anaerobic incubator with O2 concentration<6%),then reoxygenated for 6 h(incubator at 37℃and 5%CO2).In group C,1μmol/L was added for24 h hypoxia culture and 6 h reoxygenation culture.Group D was added with 10μmol/L of taxifolin,cultured in hypoxia for 24 h,and reoxygenated for 6 h.Group E was added with 100μmol/L of taxifolin,cultured for 24 h in hypoxia,and cultured for 6 h in reoxygenation.The morphological changes,survival rate,apoptotic rate,NF-κB protein expression and NF-κB inhibitory proteinα(IκB-α)protein expression in H9c2 cells were compared.Results HE staining showed that the morphology and structure of H9c2 cells in group B were abnormal,with blurred horizontal lines,a large number of spot-like necrotic lesions,obvious interstitial edema,and obvious inflammatory cell infiltration.The pathological changes of H9c2 cells in group C,D and E were significantly improved compared with group B.With the increase of dose of Taxifolin,the pathological improvement of H9c2 cells was more obvious.The survival rate and IκB-αprotein expression of H9c2 cells among five groups were the highest in group A,followed by group E,D,C and B,with statistic difference(P<0.05).The apoptosis rate and NF-κB protein expression of H9c2 cells were the lowest in group A,followed by group E,D,C and B,with statistic difference(P<0.05).Conclusion Taxifolin protects H9c2 cardiomyocytes against hypoxia-reoxygenation injury by inhibiting NF-κB signaling pathway,which may provide a new direction for prevention and treatment of myocardial ischemia-reperfusion injury.