消癥散结法对转化生长因子β_1诱导人肾小管上皮细胞表达整合素连接激酶的影响  

Influence of therapy of resolving accumulation and dissipating binds on expression of integrin-linked kinase induced by TGF-β_1 in human renal tubular epithelial cells

作　　者：刘兰英[1] 李伟 王耀献[2] 赵宗江[3] 张玉苹[3] 

机构地区：[1]北京中医药大学第三附属医院肾内科,北京100029 [2]北京中医药大学东直门医院肾内科 [3]北京中医药大学基础医学院

出　　处：《北京中医药大学学报》2012年第3期178-182,共5页Journal of Beijing University of Traditional Chinese Medicine

基　　金：高等学校博士学科点专项科研基金资助项目(No.20060026008);北京中医大学自主课题资助项目(No.JYB22-JS076)

摘　　要：目的研究消癥散结法对转化生长因子β1(TGF-β1)刺激人肾小管上皮细胞表达整合素连接激酶(ILK)的影响,探讨其在肾间质纤维化中的干预作用。方法将消癥散结法复方总方拆为补方、散方,运用血清药理学方法分别制备总方、补方、散方、盐酸贝那普利含药血清。体外培养人肾小管上皮细胞,分为6组:空白组、TGF-β1组、贝纳普利组、总方组、补方组、散方组。将5%各含药血清培养基加入体外培养的肾小管上皮细胞,除空白组外均加入TGF-β110μg/L,48 h后,提取细胞总蛋白及总RNA,分别采用免疫印迹法及逆转录-聚合酶链反应方法检测各组ILK蛋白及其mR-NA的表达。结果空白组肾小管上皮细胞胞浆内ILK及其mRNA有少量表达,TGF-β1刺激后能够诱导肾小管上皮细胞ILK及其mRNA表达显著增加(与空白组相比P<0.01);与TGF-β1组相比,贝纳普利组和总方组、散方组肾小管上皮细胞ILK及其mRNA含量明显降低(P<0.01);补方组与TGF-β1组相比差异不明显(P>0.05)。结论消癥散结法复方总方含药血清能抑制细胞ILK及其mRNA的表达,干预TGF-β1/ILK信号通路可能是其防治肾间质纤维化的机制之一。Objective To study the influence of therapy of resolving accumulation and dissipating binds on expression of integrin-linked kinase（ILK） induced by transforming growth factor β1（TGF-β1） in human renal tubular epithelial cells,and expound its intervening effect in renal interstitial fibrosis（RIF）.Methods The compound formula of whole formula with actions of resolving accumulation and dissipating binds was split to tonifying formula and dispersing formula.The medicinal serum samples of whole formula,tonifying formula,dispersing formula and benazepril hydrochloride were prepared by applying serum pharmacologic method.Human renal tubular epithelial cells were cultured in vitro and divided into blank group,TGF-β1 group,benazepril group,whole formula group,tonifying formula group and dispersing formula group.The different serum samples（5%） were added into renal tubular epithelial cells,and then all groups,except blank group,were added TGF-β1（10 ng/mL）.After 48 hours,the total protein and total RNA were extracted.The expressions of ILK protein and its mRNA were detected by using immune Western blotting method and reverse transcription-polymerase chain reaction（RT-PCR）.Results The expressions of ILK and its mRNA were less in blank group and increased significantly in TGF-β1 group（P0.01 compared with blank group）.The expressions of ILK and its mRNA decreased significantly in benazepril group,whole formula group and dispersing formula group compared with TGF-β1 group（P0.01）.The difference between tonifying formula group and benazepril group was not significant（P0.05）.Conclusion The medicinal serum of whole formula with actions of resolving accumulation and dissipating binds can inhibit the expressions of ILK protein and its mRNA,and one of the mechanisms of preventing RIF may be related to that it can intervene of signal pathway of TGF-β1/ILK.

关 键 词：消癥散结法 肾小管上皮细胞 整合素连接激酶 转化生长因子Β1 

分 类 号：R285.5[医药卫生—中药学]