Akt/GSK-3β介导高糖上调肾小管上皮细胞Snail1的表达  

Up-regulation of Snail1 expression in renal tubular epithelial cells through Akt/GSK-3β pathway under high-glucose condition

作　　者：余红[1] 石明隽[1] 肖瑛[1] 刘瑞霞[1] 王圆圆[1] 郭兵[1] 张国忠[1] 

机构地区：[1]贵阳医学院病理生理学教研室,贵州贵阳550004

出　　处：《中国病理生理杂志》2012年第12期2222-2226,共5页Chinese Journal of Pathophysiology

基　　金：贵州省科技厅资助项目(No.2008-3042)

摘　　要：目的:观察高糖对原代肾小管上皮细胞Snail1和蛋白激酶B/糖原合成酶激酶3β(Akt/GSK-3β)信号通路的影响,探讨糖尿病肾病时Snail1表达的调节机制。方法:原代培养大鼠肾小管上皮细胞(RTECs),随机分为正常糖对照组、高渗组和高糖组。Western blotting检测不同处理组RTECs不同培养时点(30 min、2 h、12 h、24h、48 h和72 h)Snail1、Akt1、GSK-3β、磷酸化Akt(p-Akt,Ser473)和磷酸化GSK-3β(p-GSK-3β,Ser9)蛋白的水平。RT-PCR检测Snail1、Akt1和GSK3βmRNA的表达。RTECs以磷脂酰肌醇3-激酶(PI3K)抑制剂LY294002(25μmol/L)预处理50 min,再与高糖共同培养24 h,Western blotting检测上述指标蛋白的表达。结果:与正常糖对照组比较,高糖组RTECs Snail1和Akt1蛋白和mRNA的表达上调,p-Akt及p-GSK-3β蛋白表达增加,但总GSK-3β蛋白和mRNA表达无变化。以LY294002处理后,高糖组RTECs Snail1、p-Akt及p-GSK-3β蛋白表达水平较未处理高糖组明显下降,但LY294002不影响总Akt1和GSK-3β蛋白表达。结论:Akt/GSK-3β可能介导了高糖诱导的RTECs锌指转录因子Snail1的表达上调。AIM ： To examine the effects of high glucose （HG） on the expression of Snail1 and protein kinase B （Akt）/glycogen synthase kinase 3β （ GSK - 3β） in primary renal tubular epithelial cells （RTECs）. METHODS ： The primary RTECs were randomly treated with normal glucose, high glucose or D - mannitol for 30 min - 72 h. RT - PCR and Western blotting were used to observe the expression of Snail1, Akt and GSK - 3β at mRNA and protein levels in these ceils. The primary cultured RTECs were pretreated with LY294002 （a PI3K inhibitor, 25 μmol/L） to observe the specific inhibitory effects of phosphatidylinositol 3 -kinase （PISK） on HG -induced expression of Snail1 protein. RESULTS： Treatment of RTECs with HG resulted in increased mRNA and protein levels of Snail1, Aktl, and phosphorylation of Akt and GSK-3β. LY294002 blocked the HG- induced up- regulation of p- Akt, p- GSK- 3β and Snail1 expression at protein level, but no effect of LY294002 was seen on the total protein expression of Aktl and GSK - 3β. HG did not affect the expression of GSK- 3β at mRNA and protein levels. CONCLUSION： HG- induced up -regulation of Snail1 may be regulated by Akt/GSK- 3β pathway in RTECs.

关 键 词：Snail1蛋白 蛋白激酶B 糖原合成酶激酶3Β 糖尿病肾病 大鼠 

分 类 号：R363[医药卫生—病理学]