黄芪多糖对硫酸吲哚酚诱导的心肌细胞损伤的保护作用

Protective Effect of Astragalus Polysaccharide on Myocardial Cell Injury Induced by Indolophenol Sulfate

目的:研究黄芪多糖对硫酸吲哚酚诱导的大鼠心肌细胞(H9c2)损伤的保护作用。方法:体外培养H9c2细胞,实验分为空白对照组、硫酸吲哚酚组、硫酸吲哚酚+黄芪多糖不同浓度组。CCK-8检测各组细胞的细胞活力,荧光分光光度计检测各组细胞ROS水平,利用RT-qPCR检测各组细胞中的Caspase-3、Caspase-9的mRNA表达水平,western blot检测各组细胞Caspase-3,Caspase-9蛋白表达水平。结果:与空白对照组比较,硫酸吲哚酚组能够显著降低H9c2的细胞活力,同时增加细胞中ROS的含量(P<0.01)。与硫酸吲哚酚组比较,硫酸吲哚酚+不同浓度黄芪多糖组,细胞活力逐渐上升,ROS含量逐渐下降(P<0.05)。此外,与硫酸吲哚酚组相比,硫酸吲哚酚+不同浓度黄芪多糖组能够明显下调caspase-3、caspase-9的mRNA及蛋白表达水平(P<0.01)。结论:硫酸吲哚酚可能通过引起氧化应激从而诱导H9c2细胞损伤,而黄芪多糖可能通过降低ROS水平,以及降低细胞中Caspase-3、Caspase-9基因及蛋白表达水平,从而改善硫酸吲哚酚对H9c2细胞的损伤。

Objective:To investigate the protections of astragalus polysaccharide on the injury of rat cardiomyocytes(H9 c2)induced by Indole sulfate(IS).Methods:H9 c2 cells were cultured in vitro,and they were divided into control group,IS group,IS+xanthine polysaccharide 50 μg/mL group,IS+xanthine polysaccharide 100 μg/mL group and IS+xanthine polysaccharide 200 μg/mL group.The morphological changes of cells in each group were observed by inverted phase contrast microscope.By detecting The cell viability,ROS levels,the mRNA expression levels of caspase-3 and caspase-9 with RT-qPCR and caspase-3,caspase-9 protein expression levels with Western blot of each group.Results:Compared with the blank control group,the IS group significantly reduced the cell viability of H9 c2 and increased the ROS content in the cells(P<0.01).The cell viability of IS+xanthine polysaccharide 50/100/200 μg/mL groups increased gradually and the ROS content decreased gradually(P<0.05).In addition,compared with IS group,IS+xanthine polysaccharide 50/100/200 μg/ml groups can significantly down-regulate the mRNA and protein expression levels of caspase-3 and caspase-9(P<0.01).Conclusion:Different concentrations of Astragalus polysaccharide can increase the cell viability of H9 c2 cells induced by IS and reduce the content of ROS.In addition,different concentrations of Astragalus polysaccharide can decrease the mRNA and protein expression of caspase-3 and caspase-9 in H9 c2 cells induced by IS,so as to achieve the purpose of inhibiting apoptosis and ultimately protect the myocardial damage caused by IS.