丹酚酸B对TGF-β1诱导的人肾小管细胞转分化的影响  

Prevent effects of salvianolic acid B on TGF-β1-induced epithelial mesenchymal transition in HK2 cells

作　　者：芮国华[1] 潘荣华[1] 姚刚[2] 武晓春[2] 徐玲玲[2] 

机构地区：[1]溧阳市中医院肾内科,江苏溧阳213300 [2]南京医科大学第二附属医院肾内科,江苏南京210011

出　　处：《南京医科大学学报（自然科学版）》2009年第12期1685-1689,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省高校自然科学基础研究项目(08KJB320006);江苏省常州市社会发展项目(CS2006217)

摘　　要：目的:探讨丹酚酸B对转化生长因子(TGF)-β1诱导的肾小管上皮细胞—间充质细胞转分化的影响。方法:将体外培养的人近端肾小管上皮细胞系(HK-2)细胞分为3组:①对照组:未加入丹酚酸B或者TGF-β1;②TGF-β1组:在细胞培养基中加入TGF-β1(浓度为5ng/ml);③丹酚酸B+TGF-β1组:在细胞培养基中分别加入丹酚酸B和TGF-β1(浓度为5ng/ml),按丹酚酸B的浓度分为A、B、C、D4个亚组:丹酚酸B的浓度分别为A组:0.1μmol/L,B组:1μmol/L,C组:10μmol/L,D组:100μmol/L。72h后,采用倒置相差显微镜观察细胞形态学变化;采用逆转录聚合酶链反应(RT-PCR)和免疫细胞化学染色检测细胞α-平滑肌肌动蛋白(α-SMA)和E-cadherin的表达情况。结果:TGF-β1组HK-2细胞从原有典型的上皮细胞形态转变为长梭形肌成纤维细胞形态;胞浆内大量表达α-SMA,同时E-cadherind的表达明显减少;不同剂量丹酚酸B治疗可减轻TGF-β1诱导的细胞形态学改变和胞浆内α-SMA的表达,同时E-cadherind的表达得到明显恢复,且呈剂量依赖性。结论:丹酚酸B具有阻止慢性肾脏疾病进行性发展的潜能,而这一作用与其能有效阻止TGF-β1诱导的HK-2细胞转分化有关。Objective：To investigate the protective effects of salvianolic acid-B （Sal B） on the trans-differentiation of renal tubular epithelial cells and to elucidate the mechanism of Sal B on renal fibrosis. Methods：Human kidney proximal tubular cell line （HK-2） was used as the proximal tubular cell model. Cells were divided into six groups as follows：control group,transforming growth factor-β1 （TGF-β1） （5 ng/ml） group,TGF-β1 （5 ng/ml） plus Sal B （0.1 μmol/L,1 μmol/L,10 μmol/L,and 100 μmol/L） groups. Epithelialto- mesenchymal transition（EMT） was induced with 5 ng / mL of human TGF-β1. The effect of Sal B on cell morphology was observed by phase contrast microscopy,and the expressions of α-smooth muscle actin （α-SMA） and E-cadherin were measured by immunocyto- chemistry and RT-PCR. Results：Our results revealed that Sal B not only prevented the expression of α-SMA but also prohibited the decrease of the epithelial marker E-cadherin in HK-2 cells in a dose-dependent manner. Simultaneous incubation of Sal B with TGF-β1 could protect the change to the myofibroblast phenotype and restored the epithelial morphology of the HK-2 cells. Conclusion：These observations strongly suggest that Sal B is a potent inhibitor of TGF-β1-induced EMT and may be a promising agent for treating tubulointerstitial fibrosis.

关 键 词：丹酚酸B 转化生长因子-Β1 上皮细胞-间充质细胞转分化 

分 类 号：R391.11[医药卫生—基础医学]