止消通脉宁干预转化生长因子-β_1诱导人肾小管上皮细胞表型转化的研究  

Effect of Zhixiaotongmaining on Phenotype Trans differentiation of Human Renal Tubular Epithelial Cell HK-2 Induced by TGF-β_1

作　　者：杨丽霞[1] 李晓东[1] 程涛[1] 刘铜华[2] 吴丽丽[2] Margetts Peter Joseph 

机构地区：[1]甘肃省中医药研究院,甘肃兰州730050 [2]北京中医药大学,北京100029 [3]加拿大麦克马斯特大学

出　　处：《中国中医药信息杂志》2013年第1期37-39,42,共4页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：科技部国际科技合作项目(2009DFA31520)

摘　　要：目的观察止消通脉宁药物血清对转化生长因子-β1(TGF-β1)诱导人肾小管上皮细胞(HK-2)表型转化的影响,并探讨其机制。方法将HK-2细胞用含10%胎牛血清的DMEM/F12(1∶1)培养基培养并分为空白对照组、TGF-β1诱导组(TGF-β110 ng/mL)、空白血清对照组(TGF-β110 ng/mL+10%空白血清)、干预1组(TGF-β110 ng/mL+10%低剂量止消通脉宁药物血清)、干预2组(TGF-β110 ng/mL+10%中剂量止消通脉宁药物血清)、干预3组(TGF-β110 ng/mL+10%高剂量止消通脉宁药物血清)。药物干预24 h后,免疫组化检测α-平滑肌肌动蛋白(α-SMA)和E-钙黏蛋白(E-cadherin)的表达。在24、48、72 h应用酶联免疫吸附法检测细胞培养上清中Ⅰ型胶原(ColⅠ)、Ⅲ型胶原(ColⅢ)和纤连蛋白(FN)的含量。结果正常HK-2细胞表达E-cadherin,不表达α-SMA,经TGF-β1刺激后细胞表型发生转变,E-cadherin的表达明显减弱,α-SMA表达明显增强,且ColⅠ、ColⅢ和FN的分泌明显增加,与空白对照组比较差异有统计学意义(P<0.05)。止消通脉宁药物血清干预后α-SMA表达明显减弱,E-cadherin表达明显增强;同时抑制了ColⅠ、ColⅢ和FN的分泌,与TGF-β1诱导组比较差异有统计学意义(P<0.05)。结论止消通脉宁在一定程度上能够抑制人肾小管上皮细胞表型转化,减少细胞外基质成分的分泌,具有防治肾间质纤维化的作用。Objective To observe the effect of Zhixiaotongmaining（ZXTMN） on epithelial-myofibroblast transdifferentiation of human renal tubular epithelial cells HK-2 induced by transforming growth factor-β1（TGF-β1）,and explore the mechanism.Methods The HK-2 cells were cultured by DMEM/F12（1∶1） with 10% fetal bovine serum and divided into control group,TGF-β1 group（TGF-β1 10 ng/mL）,animal serum control group（TGF-β1 10 ng/mL＋10% animal serum）,ZXTMN drug-containing serum therapy groups（TGF-β110 ng/mL＋10% low dose ZXTMN,TGF-β110 ng/mL＋10% medium dose ZXTMN,TGF-β1 10 ng/mL＋10% high dose ZXTMN）.After 24 h,α-SMA and E-cadherin expression were observed by immunohistochemistry assay.And after 24,48 and 72 h,the concentration of type Ⅰ collagen,type Ⅲ collagen and FN in culture medium supernatant were detected by ELISA assay.Results The expression of α-SMA of HK-2 cultured with TGF-β1 was much notable than the control,but significantly decreased in HK-2 cultured with TGF-β1 plus ZXTMN.The expression of E-cadherin was markedly decreased in HK-2 treated with TGF-β1,but not changed much in HK-2 treated with TGF-β1 plus ZXTMN.The secretion of type Ⅰ collagen,type Ⅲ collagen and FN were markedly inhibited in culture medium supernatant of HK-2 cells cultured with TGF-β1 plus ZXTMN compared with only TGF-β1（P〈0.05）.Conclution ZXTMN can prevent and treat renal fibrosis by inhibiting epithelial-myofibroblast transdifferentiation of HK-2 cell induced by TGF-β1,and reducing the synthesis and secretion of extracellular matrix.

关 键 词：止消通脉宁 转化生长因子-β1 人肾小管上皮细胞 表型转化 Ⅰ型胶原 Ⅲ型胶原 纤连蛋白 

分 类 号：R285.5[医药卫生—中药学]