GSK-3β在高糖环境下足细胞转分化效应中的作用

摘要

目的 探讨糖原合成酶激酶3β(GSK-3β)在高糖环境下足细胞转分化效应中的作用.方法 用含不同浓度葡萄糖的1640培养基处理足细胞36 h ,采用Western blot与间接免疫荧光的方法检测足细胞表面标记蛋白nephrin、podocin和间充质细胞表型标志物α平滑肌肌动蛋白(α-SMA)、Fibronectin的表达 ;用 Transwell小室检测不同处理组足细胞清蛋白流入量的变化.用GSK-3β激酶检测试剂盒检测高糖环境下足细胞GSK-3β表达量及活性的变化 ;应用GSK-3βsiRNA干扰高糖组GSK-3β后检测足细胞表型及功能变化.结果 足细胞标记蛋白nephrin、podocin的蛋白表达水平随葡萄糖浓度的升高呈剂量依赖性下调(P＜0 .05) ,间充质标记蛋白α-SMA的表达水平随葡萄糖浓度的升高呈剂量依赖性上调(P＜0 .05);清蛋白流入量随葡萄糖浓度的升高呈剂量依赖性上调(P＜0 .05).高糖组足细胞GSK-3β表达量增多(P＜0 .05).GSK-3βsiRNA处理组与对照组相比足细胞表面标记蛋白nephrin、podocin表达量增多 ,间充质标记蛋白α-SM A表达减少.结论 足细胞在高糖环境下发生表型改变与功能受损 ;GSK-3β参与高糖环境下足细胞的转分化过程.%Objective To study the action of GSK-3βin podocyte transdifferentiation effecundethe high glucose condition . MethodPodocytewere treated in RPMI-1640 medium with differenconcentrationof glucose fo36 h .The expressionof neph-rin ,podocin ,α-Smand Fibronectin were detected by both Western bloand indirecimmunofluorescence analysi.Athe same time ,the change of albumin inflow amounof podocytein differentreatmengroupwadetected by using the Transwell cham-be.The changeof expression amounand activity of GSK-3βin high glucose condition were detected by using the GSK-3 activity assay ki.The phenotypiand functional changeof podocytewere detected aftedisturbing GSK-3βby GSK-3βsiRNin the high glucose group .ResultThe expression levelof nephrin and podocin protein were down-regulated with the increase of the glucose concentration in dose-dependenmanner(P＜0 .05) ,and the expression level of α-Smwaup-regulated with the increase of the glucose concentration in dose-dependenmanner(P＜0 .05);the albumin inflow amounof podocytewaup-regulated with the in-crease of the glucose concentration in dose-dependenmanner(P＜0 .05) .The expression amounof GSK-3βin the high glucose group waincreased(P＜0 .05) .Compared with the control group ,the expression amounof nephrin and podocin aftehigh glucose treatmenin the GSK-3β siRNtreatmengroup waincreased ,while the expression of α-Smwadecreased .Conclusion The high glucose condition could induce the phenotypichange and functional impairmenof mice podocyte;GSK-3βparticipatein the epithelial-mesenchymal transdifferentiation procesof podocyte undethe high glucose condition .