槲皮黄酮通过AMPK信号通路保护营养缺乏心肌H9c2细胞的损伤

Quercetin protects H9c2 cells against nutritional stress-induced injury via AMPK signalling

目的研究槲皮黄酮对营养缺乏心肌H9c2细胞损伤的保护作用,及其对AMP依赖的蛋白激酶(AMPK)信号通路的影响。方法体外培养H9c2细胞,并将其分为对照组、营养缺乏组(NS组)、槲皮黄酮组(NS+Qu组)、Compound C组(NS+CC组)、Compound C+槲皮黄酮组(NS+CC+Qu组)。CCK8法和Hoechst染色法检测H9c2细胞增殖活性,流式细胞术检测H9c2细胞凋亡率和线粒体膜电位,ATP测定分析H9c2细胞中ATP含量,Western blot分析H9c2细胞中剪切型半胱天冬酶3(Cleaved caspase-3)、PTEN诱导假定激酶1(PINK1)、线粒体融合蛋白2(MFN2)和磷酸化AMP依赖的蛋白激酶(p-AMPK)蛋白的含量。结果与对照组比较,NS组H9c2细胞增殖活力明显降低(P<0.05),细胞凋亡率和促凋亡蛋白Cleaved caspase-3的含量明显升高(P<0.05),ATP含量、线粒体膜电位(AMRE)和线粒体自噬蛋白PINK1的含量明显降低(P<0.05)。与NS组比较,NS+Qu组H9c2细胞增殖活力明显升高(P<0.05),细胞凋亡率和Cleaved caspase-3蛋白的含量明显降低(P<0.05),ATP含量、AMRE值、PINK1和p-AMPK蛋白的含量明显升高(P<0.05)。与NS+Qu组比较,NS+CC+Qu组H9c2细胞增殖活力及AMPK磷酸化水平明显降低(P<0.05)。结论槲皮黄酮可通过调节AMPK信号通路活性改善营养缺乏诱导的心肌H9c2细胞损伤。

Aim To investigate the protective effects of quercetin on H9c2 cells injury induced by nutritional deprivation,and observe its effect on adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)signalling pathway.Methods H9c2 cells were cultured in vitro and divided into five groups:control group,nutritional stress group(NS group),quercetin group(NS+Qu group),compound C group(NS+CC group),and quercetin+compound C group(NS+CC+Qu group).The viability of H9c2 cells was determined by CCK8 assay and Hoechst staining.The apoptosis and mitochondrial membrane potentials of H9c2 cells were measured using flow cytometry.ATP content in H9c2 cells were observed.Western blot was used to detect the expression levels of Cleaved caspase-3,PTEN induced putative kinase 1(PINK1),mitofusin2(MFN2)and p-AMPK in H9c2 cells.Results Compared with control group,the viability of H9c2 cells in NS group was significantly decreased(P<0.05),the apoptotic rate and Cleaved caspase-3 expression were greatly increased(P<0.05),ATP level,mitochondrial membrane potentials(AMRE)and PINK1 expression were significantly decreased(P<0.05).Compared with NS group,the viability of H9c2 cells in NS+Qu group was significantly increased(P<0.05),the apoptotic rate and Cleaved caspase-3 expression were greatly decreased(P<0.05),ATP level,mitochondrial membrane potentials(AMRE)and PINK1 expression were significantly increased(P<0.05).Compared with NS+Qu group,the viability of H9c2 cells and p-AMPK expression level in NS+CC+Qu group was significantly decreased(P<0.05).Conclusion Quercetin can improve the injury of H9c2 cells via regulating AMPK signaling pathway.