氧化苦参碱抑制高糖诱导的大鼠肾小管上皮-间充质转化及其机制研究  

Inhibitory effect of oxymatrine on high glucose-induced rat renal tubular epithelial-mesenchymal transition

作　　者：刘丽荣[1] 李霜[2] 王圆圆[2] 石明隽[2] 肖瑛[2] 郭兵[2] 

机构地区：[1]贵阳医学院医学检验学院临床生化教研室,贵州贵阳550004 [2]贵阳医学院基础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》2013年第12期2152-2159,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81160094);贵州省中药现代化科技产业研究开发专项项目(黔科合中药字[2012]5037号)

摘　　要：目的:探讨氧化苦参碱(OM)对高糖诱导的大鼠肾小管上皮-间充质转化(EMT)的抑制作用及其可能机制。方法:体外培养大鼠近端肾小管上皮NRK52E细胞,随机分为:对照组、高糖组、高糖+OM不同浓度组和高糖+0.50 g/L OM动态观察组。采用real-time PCR和Western blotting方法检测转化生长因子β1(TGF-β1)、Smad7、α-平滑肌肌动蛋白(α-SMA)、E-钙黏素(E-cadherin)mRNA和蛋白的表达。结果:(1)与对照组相比,高糖组TGF-β1、α-SMA mRNA和蛋白表达水平均进行性增高,Smad7蛋白表达进行性降低,E-cadherin mRNA和蛋白表达进行性降低,呈时间依赖性(P<0.05),而Smad7 mRNA表达进行性增高(P<0.05);(2)与高糖组相比,高糖+OM不同浓度组随OM剂量增加,TGF-β1、α-SMA mRNA和蛋白表达均逐渐降低,Smad7蛋白表达水平逐渐增高,Ecadherin mRNA和蛋白表达水平逐渐增高,且呈剂量依赖性(P<0.05),而Smad7 mRNA表达无明显差异;(3)与高糖组相比,高糖+0.50 g/L OM动态观察组TGF-β1、α-SMA mRNA和蛋白表达持续降低,Smad7蛋白表达持续增高,E-cadherin mRNA和蛋白表达持续增高(P<0.05),而Smad7 mRNA表达无明显差异。结论:OM可抑制高糖诱导的NRK52E细胞发生EMT,其机制可能与OM下调TGF-β1表达及上调Smad7蛋白表达,进而抑制TGF-β1/Smads信号通路的致纤维化效应有关。AIM ： To investigate the inhibitory effect of oxymatrine （OM） on high glucose-induced rat renal tu- bular epithelial-mesenchymal transition （EMT）. METHODS： The rat renal tubular epithelial NRK52E cells were cultured in vitro. The ceils were divided into control group, high glucose group, high glucose ＋ different concentrations of OM groups and high glucose ＋ 0.50 g/L OM dynamic observation group. The expression of TGF-β1, Smad7, α-SMA and E-cadhefin at mRNA and protein levels was detected by real-time PCR and Western blotting. The viability of NRK52E cells was deter- mined by MTT assay. RESULTS： （1） Compared with control group, the expression of TGF-β1 and α-SMA at mRNA and protein levels in high glucose group gradually increased, and Smad7 protein and E-cadherin mRNA and protein gradually reduced, but the mRNA expression of Smad7 gradually increased. （2） Compared with high glucose group, as increases in OM doses, the expression of TGF-β1 and α-SMA at mRNA and protein levels in high glucose ＋ different concentrations of OM groups gradually reduced, and Smad7 protein and E-cadherin mRNA and protein gradually increased, but the mRNA expression of Smad7 had no significant change. （3） Compared with high glucose group, the expression of TGF-β1 and α- SMA at mRNA and protein levels was significantly reduced, the expression of E-cadherin at mRNA and protein levels significantly increased, and the protein expression of Smad7 significantly increased, but the mRNA expression of Smad7 had no significant change in high glucose ＋ O. 50 g/L OM dynamic observation group. CONCLUSION： In NRK52E cells, oxymatrine inhibits high glucose induced EMT by down-regulating TGF-β1 and up-regulating Smad7, thus preventing the fi- brosis effect of TGF-β1/Smads signaling.

关 键 词：氧化苦参碱 NRK52E细胞 转化生长因子Β1 SMAD7蛋白 上皮-间充质转化 

分 类 号：R363.2[医药卫生—病理学]