张昌志,石明隽,王圆圆,刘丽荣,肖瑛,石春花,严瑞,郭兵
ZHANG Changzhi,SHI Mingjun,WANG Yuanyuan,LIU Lirong,XIAO Ying,SHI Chunhua,YAN Rui,GUO Bing

• 1:贵阳医学院病理生理学教研室

摘要(Abstract)：

目的:探讨丹酚酸B对高糖诱导肾小管上皮细胞分化的影响及可能机制。方法:体外培养大鼠肾小管上皮细胞NRK52E随机分为正常糖(NG)组、高渗(HM)组、高糖(HG)组和高糖+Sal B(HB)组,分别培养24 h;采用免疫酶细胞化学染色对NRK52E细胞进行鉴定,MMT法检测丹酚酸B对细胞生长活性的影响,免疫荧光细胞化学和Western blotting检测各组TGF-β1、E-caderin、α-SMA和Col-Ⅳ蛋白的表达。结果:与NG组相比较,HG组肾小管上皮细胞中TGF-β1、α-SMA和Col-Ⅳ蛋白表达增多(P<0.05),而E-cadherin蛋白表达降低(P<0.05);与HG组比较,HB组肾小管上皮细胞中TGF-β1、α-SMA和Col-Ⅳ蛋白表达减少(P<0.05),而E-cadherin蛋白表达增多(P<0.05)。结论:丹酚酸B对高糖诱导肾小管上皮细胞的上皮-间充质转化(EMT)有明显抑制作用,其机制可能是通过抑制TGF-β1的表达,改善了肾小管纤维化病变的发生发展。
Objective: To investigate the effect of Salvianolic acid B( Sal B) on the high-glucose induced epithelial-mesenchymal transition( EMT) in renal tubule epithelial cells and the possible mechanism. Methods: NRK52 E cells,the rat renal tubular epithelial cells,were cultured in vitro. The cells were randomly divided into normal glucose group( group NG),high mannitol group( group HM),high glucose group( group HG) and high glucose and Sal B group( group HB),and each group was cultured for 24 h. Immunocytochemistry was used to identify the renal tubule epithelial cells. MTT was adopted to the effect of Salvianolic acid B on cell growth activity. Immunofluorescence cytochemistry staining was used to assess the protein expression of E-cadherin and α-SMA. Western blotting was used to detect the protein expression of TGF-β1,E-caderin,α-SMA and Col-Ⅳ under different conditions. Results: Compared with group NG,the protein expression of TGF-β1,α-SMA and Col-Ⅳ increased significantly( P < 0. 05) while the expression of E-cadherin significantly decreased( P < 0. 05) in renal tubular epithelium after treated with high glucose for 24 h. Compared with group HG,in group HB,Sal B decreased the expression of TGF-β1,α-SMA and Col-Ⅳ( P < 0. 05)and upregulated E-cadherin protein expression in renal tubular epithelium cells after treated with Sal B for 24 h( P < 0. 05). Conclusion: Sal B can significantly inhibit HG-induced expression of EMT through reducing the expression of TGF-β1,which may ameliorate occurrence and development of renal tubule fibrosis lesion.

关键词(KeyWords)： 肾小管上皮细胞;高糖培养;丹酚酸B;转化生长因子-β1;上皮-间充质细胞转化
renal tubular epithelial cells;high-glucose;Salvianolic acid B;transforming growth factor betal;epithelial-mesenchymal transition

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(编号:81360116);; 贵州省社会发展攻关项目[编号:黔科合SY字(2012)3088号];; 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]

作者(Author): 张昌志,石明隽,王圆圆,刘丽荣,肖瑛,石春花,严瑞,郭兵
ZHANG Changzhi,SHI Mingjun,WANG Yuanyuan,LIU Lirong,XIAO Ying,SHI Chunhua,YAN Rui,GUO Bing

DOI: 10.19367/j.cnki.1000-2707.2015.04.003

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