Snail1 siRNA对高糖诱导肾小管上皮细胞表型转变的影响  

Effects of Snail1 siRNA on tubular epithelial-to-mesenchymal transition induced by high glucose

作　　者：方开云[1,2] 石明隽[1] 肖瑛[1] 桂华珍[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 [2]贵州省人民医院麻醉科,贵州贵阳550002

出　　处：《中国病理生理杂志》2009年第12期2424-2429,共6页Chinese Journal of Pathophysiology

基　　金：教育部高等学校博士学科点专项科研基金资助项目(No.20060660001);贵州省科技厅攻关资助项目(No.2008-3041)

摘　　要：目的:观察Snail1 siRNA对高糖诱导的肾小管上皮细胞向间充质细胞转变(TEMT)的影响。方法:原代培养肾小管上皮细胞分为5组:(1)对照组(含糖5.5mmol/L);(2)高糖组(含糖25mmol/L);(3)Snail1 siRNA处理组,转染Snail1 siRNA,6h后更换为高糖(含糖25mmol/L)培养;(4)control siRNA处理组,转染control siRNA作为siRNA阴性对照,6h后换为高糖(含糖25mmol/L)培养;(5)高渗组(含D-manntio19.5mmol/L);72h后收集细胞,用Western blotting和半定量RT-PCR检测Snail1、TGF-β1、α-平滑肌肌动蛋白(α-SMA)、vimentin和E-cadherin蛋白和mRNA表达。结果:与高糖组比较,肾小管上皮细胞转染Snail1 siRNA后,Snail1 mRNA和蛋白表达水平分别下降62%和68%(P<0.01)。同时,Snail1 siRNA处理组α-SMA和vimentin蛋白和mRNA表达显著下调(P<0.01),而E-cadherin蛋白和mRNA表达显著上调(P<0.01)。结论:Snail1参与了高糖诱导TEMT的调节。AIM： To explore the effect of Snaill siRNA on high - glucose induced tubular epithelial - to - mesenchymal transition （TEMT）. METHODS： Subconfluent renal tubular epithelial cells were incubated in serum -free DMEM for 24 h to arrest and synchronize the cell growth. Then cells were treated with normal glucose （5.5 mmol/L D - glucose） or high glucose （25 mmol/L D - glucose） for 72 h. Meanwhile 19. 5 mmol/L D - manntiol was used as high osmotic control. Snaill siRNA was transfected into tubular epithelial cells. In parallel, cells were transfected with non - specific siRNA which served as the control data sets. Cells were then treated with 25 mmol/L D - glucose for 72 h. RNA and cell lysates were collected to determine the protein and mRNA levels of Snaill, TGF - β1, α - SMA, vimentin and E - cadherin. RESULTS： Transfection caused the decreases in Snaill at mRNA and protein levels by 62% and 68% respectively as compared to those in untransfected cells cultured in high glucose medium. Western blotting exhibited that Snaill siRNA transfection restored E - cadherin protein expression by 61% compared to that in high - glucose - treatment cells, whereas it inhibited high - glucose - induced induction of α - SMA protein by 58%. Similarly, RT - PCR revealed that Snaill siRNA transfection dramatically suppressed the high - glucose - induced mRNA expressions of α - SMA and vimentin by 72% and 61%, respectively, while E -cadherin mRNA increased by 53%. CONCLUSION： Our study provides direct evidence that Snaill is able to control TEMT.

关 键 词：SNAIL1 SIRNA 肾小管上皮细胞向间充质细胞转变 高糖 

分 类 号：R363[医药卫生—病理学]