芍药苷对高糖诱导肾小球系膜细胞凋亡的影响及机制探究

Effect of paeoniflorin on apoptosis of mesangial cells induced by high glucose and its mechanism

目的观察芍药苷对高糖诱导肾小球系膜细胞凋亡的影响,并探讨其作用机制。方法将SV40 MES 13小鼠肾小球系膜细胞分为5组:对照组、高糖组(60 mmol·L^(-1))和芍药苷组(2.5、10、30μmmol·L^(-1)),Hoechst 33258观察细胞形态,Annexin V-FITC/PI双标记检测细胞凋亡,JC-1染色检测细胞线粒体膜电位,透射电镜观察细胞线粒体形态,Western blot检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关蛋白X(Bcl-2 associated X protein,Bax)、半胱氨酸天冬氨酸蛋白水解酶-3(cysteinyl aspartate specific protease-3,Caspase-3)、细胞色素c(cytochrome c,Cyt-c)、线粒体融合蛋白2(mitofusion 2,MFN2)和线粒体动力相关蛋白(dynamin-related protein 1,DRP1)的表达水平。结果与对照组比较,高糖组细胞凋亡率明显升高(P<0.05),线粒体空泡化、嵴融合现象严重,线粒体膜电位明显降低(P<0.05),Bax、Caspase-3、Cyt-c、DRP1蛋白表达明显升高(P<0.05),Bcl-2和MFN2表达明显降低(P<0.05),而经芍药苷预处理后,高糖诱导所引起的上述变化均得到缓解,且呈现出芍药苷浓度依赖性。结论芍药苷预处理肾小球系膜细胞,可减轻高糖诱导的细胞凋亡,其作用机制可能与保护细胞线粒体结构和功能有关。

Objective To observe the effect and mechanism of paeoniflorin on apoptosis of mesangial cells induced by high glucose.Methods SV40 MES 13 of mouse mesangial cells were divided into 5 groups:control,high glucose(60 mmol·L^(-1)),paeoniflorin(2.5,10 and 30μmol·L^(-1)).Cell morphology was observed by Hoechst 33258.Cell apoptosis was detected by Annexin V-FITC/PI double labeling.JC-1 was used to detect mitochondrial membrane potential and transmission electron microscopy was used to observe the morphology of mitochondria.The protein expressions of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cysteinyl aspartate specific protease-3(Caspase-3),cytochrome c(Cyt-c),mitofusion 2(MFN2)and dynamin-related protein 1(DRP1)were detected by Western blot.Results Compared with the control group,the apoptosis rate was significantly increased(P<0.05);mitochondrial vacuolation and cristae fusion were serious;mitochondrial membrane potential was significantly reduced(P<0.05);the proteins expressions of Bax,Caspase-3,Cyt-c and DRP1 were significantly increased(P<0.05);Bcl-2 and MFN2 expressions were significantly decreased(P<0.05)in the high glucose group.However,after pretreatment with paeoniflorin,the above changes induced by high glucose were alleviated,which was related to the concentration dependence of paeoniflorin.Conclusion Pretreatment of mesangial cells with paeoniflorin can reduce the apoptosis induced by high glucose,and its mechanism may be related to the protection of mitochondrial structure and function.