加味当归补血汤对肾小管上皮细胞TGF-β_1/SMADs信号转导通路的影响  

Effect of Jia Wei Dang Gui Bu Xue Tang on TGF-β_1/SMADs Signaling Pathway in Renal Tubular Epithelia Cells

作　　者：张敏鸥[1] 卜爽剡 王永钧[1] 朱晓玲[1] 杨汝春[1] 王豫巍[3] 

机构地区：[1]浙江省杭州市中医院肾内科,杭州310007 [2]浙江省东阳市人民医院内科,东阳322100 [3]浙江中医药大学,2004级研究生杭州310053

出　　处：《中国中西医结合肾病杂志》2007年第9期503-506,F0002,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：浙江省科技厅基金资助项目(No.021110685)

摘　　要：目的：观察加味当归补血汤对转化生长因子β1（TGF-β1）刺激的小鼠肾小管上皮细胞Smad3、Smad4、Smad7信号通路的影响，探讨该方防治肾间质纤维化的细胞分子生物学机制。方法：原代培养BalB／C小鼠肾小管上皮细胞．用TGF-β（1ng／ml）刺激24h，加味当归补血汤及洛汀新含药血清干预。共分6组：正常组、模型组、加味当归补血汤低中高剂量组（含药血清浓度分别为2．5％、5％、10％）、洛汀新组，观察各组细胞形态学变化，检测细胞Smad3、Smad4、Smad7的mRNA和蛋白质表达情况（PT—PCR、Western-Blotting）。结果：（1）TGF-β1刺激后，肾小管上皮细胞部分形态发生变大，伸长，呈梭形，经中药加味当归补血汤和洛汀新干预后，细胞形态又恢复正常；（2）TGF-β1刺激肾小管上皮细胞后，Smad3、Smad4 mRNA和蛋白质表达显著增加，Smad7 mRNA和蛋白质则显著减少（P〈0、05～0、01）；（3）各浓度加味当归补血汤能显著下调异常增高的Smad3，上调Smad7的基因和蛋白质表达水平（P〈0．05～0．01），能显著下调Smad4基因表达。结论：加味当归补血汤防治肾间质纤维化可能与调控肾小管上皮细胞TGF-β1／Smads信号转导途径相关。Objective：To study the effect and molecule mechanism of Jia Wei Dang Gui Bu Xue Tang （JDBT） on signal protein Smad3/4/-/in renal tubular epithelial cells （RTEC） stimulated by TGF- β1-Methods： Primary- cultured mouse RTEC was stimulated by TGF - 61 and treated by herb serum of JDBT/Lotinsin for 24 hours. There were 6 groups： normal, model, 2.5 %/5 % / 10% JDBT herb serum group and Lotensin serum group. In all groups, the end serum concentration was 10%. The morphology of RTEC was observed under inversion microscope. The expression of Smad3/4/-/mRNA and protein production of RTEC were measured by RT - PCR and Western Blotting, respectively. Results： （ 1 ） The morphology of RTEC in model group became bigger and elongated with a fusiform shape. The morphology of RTEC in JDBT herb/Lotensin serum groups was in cobble shape as natural RTEC. （2） The expression of Smad3/4 mRNA and protein production were significantly increased and that of smad7 decreased in model group （P〈0.05～0.01）. （3） High expression of smad3/4 mRNA and smad3 protein production could be significantly down - regulated and smad7 mRNA and protein production up - regulated by JDBT herb serum （P〈 0.05～0.01 ）. Conclusion： JDBT may prevent renal interstitial fibrosis through regulating TGF - β1/Smads signal protein in RTEC.

关 键 词：加味当归补血汤 肾小管上皮细胞 转化生长因子Β1 SMADS 

分 类 号：R285.5[医药卫生—中药学]