化瘀降浊汤对转化生长因子-β1/母系同源物家族信号通路相关蛋白的影响  

Effects of Huayu Jiangzhuo Decoction on TGF-β1/Mothers against Decapentaplegic Homolog Signal Pathway Relevant Proteins

作　　者：尹学来[1] 宋竖旗[1] 庞然[1] 吕济源 卢建新[1] YIN Xue-lai;SONG Shu-qi;PANG Ran;LV Ji-yuan;LU Jian-xin(Urology of Guang’anmen Hospital, China Academy of Chinese Medicial Sciences, Beijing 100053, China)

机构地区：[1]中国中医科学院广安门医院泌尿外科,北京100053

出　　处：《中国中医药信息杂志》2016年第8期54-58,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81250032);北京市自然科学基金(7132226;7112119;7112118);北京中医药科技发展基金项目(JJ2013-63)

摘　　要：目的观察化瘀降浊汤对转化生长因子(TGF)-β1诱导HK-2细胞发生上皮间质转分化(EMT)过程TGF-β/母系同源物家族Smad通路中关键因子E-钙黏蛋白(E-cadherin)、Smad3、蜗形蛋白(Snail)的影响,探讨其保护肾功能的作用机制。方法 MTT法测定不同浓度化瘀降浊汤对TGF-β1诱导HK-2的细胞增殖抑制率,计算半数抑制率(IC50)。Western blot和RT-PCR检测E-cadherin、Smad3、Snail蛋白和基因表达,间接免疫荧光检测E-cadherin蛋白表达。结果化瘀降浊汤在一定浓度范围内抑制TGF-β1诱导的HK-2细胞增殖,呈现出一定的量效关系,IC_(50)为472.017μg/m L。Western blot和RT-PCR检测结果显示,化瘀降浊汤中、高剂量组E-cadherin、Smad3、Snail蛋白和m RNA表达与诱导组比较,差异有统计学意义(P<0.05,P<0.01);免疫荧光检测结果显示,化瘀降浊汤能促进E-cadherin蛋白表达,与Western blot分析结果一致。结论化瘀降浊汤可能通过调节HK-2细胞TGF-β/Smad信号途径中的关键蛋白达到抑制甚至逆转肾小管EMT,从而对肾小管间质纤维化起到保护作用。Objective To investigate the effects of Huayu Jiangzhuo Decoction on the progress of TGF-β1inducing HK-2 to generate epithelial-mesenchymal transition (EMT) and its crucial factors E-cadherin, Smad3, andSnail in the pathway of TGF/β-mothers against decapentaplegic homolog (Smad); To discuss the mechanism of actionfor protecting renal function. Methods MTT method was used to detect the inhibition ratio of Huayu JiangzhuoDecoction for TGF-β1 induced HK-2 cell proliferation. IC50 was counted. E-cadherin, Smad3, and Snail protein andgenetic expression were detected by Western blot and RT-PCR. E-cadherin protein expression was detected by indirectimmunofluorescence. Results Huayu Jiangzhuo Decotion can inhibit the TGF-β1 induced HK-2 cell proliferation tosome extent, with dose-effect relationship and IC50=472.017 μg/mL. The results of Western blot and RT-PCR showedthat the protein E-cadherin, Smad3, and Snail and mRNA expression in medium- and high-dose Huayu JiangzhuoDecotion groups had statistical significance compared with induced group (P<0.05, P<0.01). The results ofimmunofluorescence showed that Huayu Jiangzhuo Decoction could promote the protein expression of E-cadherinmRNA, and were in accordance with the results of Western blot (P<0.01).Conclusion Huayu Jiangzhuo Decoctioncan inhibit the TGF-β1 induced HK-2 cell proliferation, reverse the progress of EMT, and conduct protective effect oninterstitial fibrosis of kidney tubules by regulating its crucial protein E-cadherin, Smad3, and Snail in the pathway ofTGF/β-Smad.

关 键 词：HK-2 细胞 上皮间质转化 化瘀降浊汤 E-钙黏蛋白 SMAD3 蜗形蛋白 

分 类 号：R285.5[医药卫生—中药学]