益肾胶囊通过上调SIRT1抑制高糖诱导足细胞凋亡的机制研究

Mechanism of Yishen Capsule Inhibiting Podocyte Apoptosis Induced by High Glucose by Raising SIRT1

目的:探讨益肾胶囊通过调控SIRT1表达,抑制炎症反应,减轻高糖诱导的足细胞凋亡。方法:体外培养小鼠足细胞,(1)给予SIRT1及SIRT1siRNA质粒转染分为:(1)NC组(空白质粒组);(2)过表达组(SIRT1质粒);(3)si组(SIRT1siRNA质粒);(2)给予益肾胶囊干预,实验分为:(1)NG组(5.5 mmol/L葡萄糖);(2)HG组(30 mmol/L葡萄糖);(3)YS组(30 mmol/L葡萄糖+10%益肾胶囊含药血清);(4)R组(30 mmol/L葡萄糖+10 mmol/L白藜芦醇),western blot及RTPCR检测SIRT1、NF-κBp65、BCL-2、MCP-1的蛋白和mRNA表达,流式细胞检测细胞凋亡。结果:(1)SIRT1过表达组的SIRT1、Bcl-2的蛋白和mRNA表达升高(P<0.05),乙酰化NF-κBp65、MCP-1的蛋白和mRNA表达下降(P<0.05);SIRT1siRNA组的SIRT1、Bcl-2的蛋白和mRNA表达下降(P<0.05),乙酰化NF-κBp65、MCP-1的蛋白和mRNA表达升高(P<0.05)。(2)高糖组的SIRT1、Bcl-2的蛋白和mRNA表达较正常组下降(P<0.05),乙酰化NF-κBp65、MCP-1蛋白和mRNA表达升高(P<0.05)。益肾胶囊含药血清组和白藜芦醇组的SIRT1、Bcl-2的蛋白和mRNA表达升高(P<0.05),乙酰化NF-κBp65、MCP-1蛋白和mRNA表达下降(P<0.05)。(3)高糖组足细胞凋亡率较正常组高(P<0.05),益肾胶囊含药血清组和白藜芦醇组的足细胞凋亡率低于高糖组(P<0.05)。结论:益肾胶囊可能通过上调足细胞SIRT1表达,抑制足细胞炎症及凋亡,修复足细胞损伤。

Objective:To study the expression of Yishen capsule by regulating SIRT1,Suppresses the inflammatory response,attenuate high glucose-induced podocyte apoptosis.Methods:(1)In vitro culture of mouse podocytes,were divided into:(1)NC group(blank plasmid group);(2)Overexpression group(SIRT1 plasmid);(3)Si Group(SIRT1 siRNA plasmid).(2)Mouse podocytes were divided into:(1)N group(5.5 mmol/L glucose);(2)HG group(30 mmol/L glucose);(3)YS group(30 mmol/L glucose+10%Yishen Capsule Containing Serum);(4)R group(30 mmol/L glucose+10 mmol/L resveratrol).To detect protein and mRNA expression in western blot and RT-PCR,Flow cytometry was used to detect apoptosis.Results:(1)The expression of protein and mRNA in SIRT1 overexpression group increased(P<0.05),The protein and mRNA expression of acetylated NF-κBp65、MCP-1 decreased(P<0.05);SIRT1 siRNA SIRT1、Bcl-2 protein and mRNA expression decreased in the group(P<0.05),protein and mRNA expression in acetylated NF-κBp65、MCP-1 increased(P<0.05).(2)The expression of SIRT1、Bcl-2 protein and mRNA decreased in the high glucose group(P<0.05),The expression of acetylated NF-κBp65、MCP-1 protein and mRNA increased(P<0.05).The expression of SIRT1、Bcl-2 protein and mRNA in Yishen capsule containing serum group and triglyceride group increased(P<0.05),The expression of acetylated NF-κBp65、MCP-1 protein and mRNA decreased(P<0.05).(3)The apoptosis rate of podocytes in high sugar group was higher than that in normal group(P<0.05),Apoptosis rate of podocytes in Yishen capsule serum group and triglyceride group was lower than that in high glucose group(P<0.05).Conclusion:Yishen capsule may regulate the expression SIRT1 podocytes,inhibit podocyte inflammation and apoptosis,and repair podocyte injury.