党参多糖联合沉默调节蛋白4对体外人脐静脉糖尿病血管内皮细胞凋亡的影响

目的:研究党参多糖联合SIRT4对体外人脐静脉糖尿病血管内皮细胞凋亡的影响。方法:血管内皮细胞分成空白对照组(常规细胞培养液培养)、高糖模型组(30mmol/L葡萄糖处理)、转染阴性对照组(转染阴性对照载体并经30mmol/L葡萄糖处理)、转染SIRT4组(转染SIRT4过表达载体并经30mmol/L葡萄糖处理)、转染阴性+党参多糖组(转染阴性对照载体并经30mmol/L葡萄糖处理和100μg/ml党参多糖处理)、转染SIRT4+党参多糖组(转染SIRT4过表达载体并经30mmol/L葡萄糖处理和100μg/ml党参多糖处理)。MTT检测细胞增殖,流式细胞术检测细胞凋亡,ELISA检测ROS、MDA、SOD、CAT水平,JC-1染色法检测线粒体膜电位,Western blot检测C-Caspase-3蛋白和cyt-c蛋白表达。结果:与空白对照组比较,高糖模型组细胞增殖能力降低,细胞凋亡和C-Caspase-3蛋白表达增多,细胞中ROS和MDA水平升高,SOD、CAT水平下降,胞浆cyt-c蛋白增多,线粒体cyt-c蛋白减少,线粒体膜电位降低。与转染阴性对照组比较,转染SIRT4组和转染阴性+党参多糖组细胞增殖能力升高,细胞凋亡和C-Caspase-3蛋白表达减少,细胞中ROS和MDA水平降低,SOD、CAT水平升高,线粒体膜电位升高,胞浆cyt-c蛋白减少,线粒体cyt-c蛋白增多。与转染SIRT4组和转染阴性+党参多糖组比较,转染SIRT4+党参多糖组细胞增殖能力升高,细胞凋亡和C-Caspase-3蛋白表达减少,细胞中ROS和MDA水平降低,SOD、CAT水平升高,线粒体膜电位升高,胞浆cyt-c蛋白减少,线粒体cyt-c蛋白增多。结论:党参多糖联合SIRT4可抑制体外糖尿病血管内皮细胞凋亡,作用机制可能与其抑制线粒体凋亡途径有关。

Effects of Radix Codonopsis Polysaccharides Combined with Sirtuin 4 on Apoptosis of Vascular Endothelial Cells Induce by High Glucose in Vitro

Objective:To study the effect of radix codonopsis polysaccharides combined with sirtuin 4(SIRT4)on the apoptosis of vascular endothelial cells induce by high glucose in vitro.Method:Vascular endothelial cells were divided into control group,model group(high glucose treatment),vector group(transfection negative control vector,high glucose treatment),SIRT4 group(transfection SIRT4 overexpression vector,high glucose treatment),vector+Radix codonopsis polysaccharides group(transfection negative control vector,high sugar and radix codonopsis polysaccharides treatment),SIRT4+Radix codonopsis polysaccharides group(transfected with SIRT4 overexpression,high sugar and radix codonopsis polysaccharides treatment).MTT was used to detect cell proliferation,flow cytometry was used to detect apoptosis,ELISA kits were used to detect ROS,MDA,SOD,CAT levels,JC-1 staining was used to detect mitochondrial membrane potential,and Western blot was used to detect C-Caspase-3 protein expression,Cyt-c protein expression in mitochondria.Results:Compared with the control group,the cell proliferation ability of the model group decreased,apoptosis and C-Caspase-3 protein expression increased,ROS and MDA levels increased,SOD and CAT levels decreased,mitochondrial membrane potential decreased,and cyt-c in the cytoplasm increased,cyt-c decreased in mitochondria.Compared with the vector group,SIRT4 group and vector+RCPS group had increased cell proliferation ability,decreased apoptosis and C-Caspase-3 protein expression,decreased ROS and MDA levels,increased SOD and CAT levels,and increased mitochondrial membrane potential,cyt-c decreased in cytoplasm,and cyt-c increased in mitochondria.Compared with SIRT4 group and vector+Radix codonopsis polysaccharides group,SIRT4+Radix codonopsis polysaccharides group had increased cell proliferation ability,decreased apoptosis and C-Caspase-3 protein expression,decreased ROS and MDA levels in cells,increased SOD and CAT levels,and increased mitochondrial membrane,the potential increased,cyt-c decreased in cytoplasm,and cyt-c increased in mitochondria.Conclusion:Radix codonopsis polysaccharides combined with SIRT4 can inhibit the apoptosis of diabetic vascular endothelial cells in vitro,and the mechanism may be related to the mitochondrial apoptosis pathway.