糖耐康对TGF-β_1诱导的HK-2细胞Smads通路的影响  

To Explore the Effect of Tangnaikang on Smads Signaling Pathway of Human Renal Tubular Epithelial HK-2 Cell Induced by TGF-β_1

作　　者：杨丽霞[1,2] 舒畅[2] 吴丽丽[3] 孙文[3] 刘铜华[3] 

机构地区：[1]甘肃省中医药研究院,兰州730050 [2]甘肃中医学院,兰州730000 [3]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2014年第20期160-163,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(30973909);北京中医药大学创新团队项目(2011-CXTD-19)

摘　　要：目的:探讨糖耐康(TNK)含药血清对转化生长因子-β1(TGF-β1)诱导的人肾小管上皮细胞(HK-2)转分化Smad信号通路的影响。方法:将HK-2细胞用含10%胎牛血清的DMEM/F12(1∶1)培养基培养;实验分为6组:空白对照组、单纯TGF-β1诱导组(TGF-β110μg·L-1)、空白血清对照组(TGF-β110μg·L-1+10%空白血清)、TNK高浓度组(TGF-β110μg·L-1+20%糖耐康含药血清)、TNK中浓度组(TGF-β110μg·L-1+10%糖耐康含药血清)、TNK低浓度组(TGF-β110μg·L-1+5%糖耐康含药血清)。药物干预24 h后,荧光定量PCR检测TGF-β1及其Ⅰ,Ⅱ受体(TβRI,TβRⅡ)的mRNA表达,Western blot检测Smad 2、Smad 3的蛋白表达。结果:HK-2细胞经TGF-β1诱导后,TβRⅠ,TβRⅡ的mRNA表达和Smad 2,Smad 3的蛋白表达显著上升,与空白对照组相比有显著性差异(P<0.05),但经TNK含药血清干预后,其表达逐步下降,与单纯TGF-β1诱导组及TGF-β1+空白血清对照组相比有显著性差异(P<0.05)。而空白血清无此作用。结论:TNK能够调控TGF-β1诱导的人肾小管上皮细胞转分化Smad信号通路,在一定程度上具有抑制肾间质纤维化的作用。Objective： To explore the effect of Tangnaikang （TNK） on the smads signaling pathway of human renal tubular epithelial cells （HK-2） induced by transforming growth factor-β1 （TGF-β1）. Method： 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 10μg ·L^-1）, rat serum control group （TGF-β1 10g·L^-1 ＋ 10% rat serum）, TNK - containing rat serum therapy groups （TGF-β,10 μg·L^-1 ＋ 20% TNK or ＋ 10% TNK or ＋ 5% TNK）. After 24 h, the expression of transforming growth factor ]3 receptor I （TβRI） , transforming growth factor β receptor II （TβRII） mRNA were tested by fluorescence quantitatiye PCR assay, and the expression of Smad 2, Smad 3 protein by Western-blot assay. Result： The expression of TβRI, TβRII mRNA and Smad 2, Smad 3 protein of HK-2 cultured with TGF-β1 were much notable than the control, and significantly decreased in HK-2 cultured with TGF-β1 plus TNK compared with only TGF-β1 （P 〈 0.05） , but only rat serum had no such effect. Conclusion：TNK could regulate the smads signaling pathway of human renal tubular epithelial cells HK-2 induced by transforming growth factor-β1 , and could prevent the development of renal fibrosis to a certain extent.

关 键 词：糖耐康 转化生长因子β1 HK-2细胞 转化生长因子-Β1 I受体 转化生长因子-Β1 Ⅱ受体 SMAD2 SMAD3 

分 类 号：R285.5[医药卫生—中药学]