姜黄素调控PI3K/Akt信号通路抑制H9c2心肌细胞的炎症损伤

Curcumin inhibits inflammatory injury in H9c2 cardiac myocytes through regulating PI3K/Akt signaling pathway

目的探讨姜黄素对脂多糖(LPS)诱导的心肌细胞炎症损伤的保护效应,并对其作用机制进行初步研究。方法将H9c2心肌细胞分为对照组、LPS组和姜黄素干预组。除对照组外,其余各组构建LPS诱导的心肌细胞炎症损伤模型。姜黄素干预组分别以10μmol/L、20μmol/L和40μmol/L姜黄素处理细胞。培养24 h后,CCK8法检测各组细胞的存活率,ELISA法检测各组细胞分泌炎症因子IL-6和TNF-α的水平,Western blot法检测各组细胞PI3K、Akt和p-Akt蛋白表达水平。采用PI3K抑制剂LY294002(5μmol/L)预先处理细胞4 h后,加入LPS诱导细胞损伤,再以20μmol/L姜黄素干预细胞,检测各组(对照组、LPS组、LPS+抑制剂组、LPS+姜黄素组和抑制剂+LPS+姜黄素组)细胞的相对存活率、炎症因子水平及PI3K/Akt通路蛋白的表达。结果与对照组相比,LPS组细胞的相对存活率显著降低(P<0.05),细胞上清中炎症因子IL-6和TNF-α水平显著升高(P<0.05),且PI3K和p-AKT蛋白表达量显著降低(P<0.05)。与LPS组细胞相比,不同浓度姜黄素干预后,H9c2细胞相对存活率显著升高(P<0.05),细胞分泌的IL-6和TNF-α水平显著降低(P<0.05),且PI3K和p-AKT蛋白表达量显著升高(P<0.05)。与LPS+姜黄素组相比,抑制剂LY294002干预细胞后,姜黄素对LPS诱导的H9c2细胞炎症损伤的保护作用减弱,细胞存活率明显下降(P<0.05),IL-6和TNF-α水平显著升高(P<0.05),且PI3K和p-AKT蛋白表达量显著降低(P<0.05)。结论姜黄素对LPS诱导的H9c2心肌细胞炎症损伤具有保护效应,其作用机制可能与激活PI3K/Akt信号通路有关。

Objective To observe the protective effects of curcumin on lipopolysaccharide（ LPS） induced inflammatory injury in H9c2 cardiac myocytes,and preliminarily explore the potential mechanism.Methods The H9c2 cardiac myocytes were divided into the control group,LPS group and curcumin treatment groups.Except the control group,the LPS induced inflammatory injury model in cardiac myocytes was established in the other groups.The cells in the curcumin treatment groups were treated with curcumin at concentrations of 10 μmol/L,20 μmol/L and 40 μmol/L,respectively.After cultured for 24 hours,the survival rate of H9c2 cells was measured by CCK8 kit,the levels of inflammatory cytokines such as IL-6 and TNF-α were detected by ELISA,and the protein expression levels of PI3 K,Akt and p-Akt were detected by Western blot.After pre-treatment with 5 μmol/L PI3 K inhibitor（ LY294002） for 4 hours,the LPS was added into the cells to induce the injury,then the cells were treated with 20 μmol/L curcumin.The survival rate of cells,the levels of inflammatory cytokines and the expression levels of PI3 K/Akt pathway related proteins were detected in each group,including the control group,LPS group,LPS ＋ LY294002 group,LPS ＋ curcumin group and LY294002＋LPS＋ curcumin group.Results Compared with the control group,the relative survive rate of H9c2 cells was significantly decreased in the LPS group（ P〈0.05）,the IL-6 and TNF-α levels in the cell supernatant were increased significantly（ P〈0.05）,and the protein expressions of PI3 K and p-Akt were significantly decreased（ P〈0.05）.Compared with the LPS group,after treatment with curcumin at different concentrations,the relative survive rate of H9c2 cells was significantly increased（ P〈0.05）,the IL-6 and TNF-α levels were significantly decreased（ P〈0.05）,and the protein expressions of PI3 K and p-Akt were significantly increased（ P〈0.05）.Compared with the LPS ＋ curcumin group,after treatment with LY294002,the protective effects of curcumin on LPS induced inflammatory injury in H9c2 cardiac myocytes were significantly reduced,the cell survival rate was obviously decreased in LY294002 ＋ LPS ＋ curcumin group（ P〈0.05）,the levels of IL-6 and TNF-α were significantly increased（ P〈0.05）,and the protein expressions of PI3 K and p-Akt were significantly decreased（ P〈0.05）.Conclusion Curcumin exerts protective effects on LPS induced inflammatory injury in H9c2 cardiac myocytes,and its mechanism may be associated with activating PI3 K/Akt signaling pathway.