六味地黄丸含药血清及梓醇对肾小管上皮细胞HK-2转化生长因子β1/Smad通路的影响  

Effects of Chinese herbal medicine Liuwei Dihuang Pill and its main monomer catalpol on transforming growth factor-β1/Smad pathway in HK-2 cells

作　　者：刘煜敏[1] 张悦[2] 郝艳鹏[1] 李志杰[1] 

机构地区：[1]上海中医药大学科技实验中心,上海201203 [2]上海中医药大学基础医学院,上海201203

出　　处：《中西医结合学报》2011年第7期783-788,共6页Journal of Chinese Integrative Medicine

基　　金：上海市教育委员会重点学科(第五期)资助项目(No.J50301);上海高校创新团队建设项目

摘　　要：目的:观察六味地黄丸含药血清及其主要单体梓醇对肾小管上皮细胞株HK-2转化生长因子-β1(transforming growth factor-β1,TGF-β1)/Smad信号通路的影响。方法:制备大鼠六味地黄丸含药血清,采用四甲基偶氮唑盐法和乳酸脱氢酶释放实验确定药物最适作用浓度。六味地黄丸含药血清和梓醇作用于TGF-β1刺激(或不刺激)的HK-2细胞,并设肝细胞生长因子(hepatocyte growthfactor,HGF)为阳性对照,采用蛋白质印迹法观察Smad通路分子Smad2、Smad7和Ski相关活性蛋白N(Ski-related novel protein N,SnoN)蛋白表达的情况。结果:与HGF作用相似,10%浓度六味地黄丸含药血清可抑制TGF-β1诱导的HK-2细胞Smad2蛋白磷酸化并促进核转录抑制因子SnoN的表达。梓醇亦可显著抑制HK-2细胞Smad2蛋白磷酸化。六味地黄丸含药血清及梓醇对HK-2细胞Smad7蛋白表达未见明显调控作用。结论:在TGF-β1刺激HK-2细胞模型中,六味地黄丸含药血清拮抗Smad通路活化的作用与HGF相似,梓醇是六味地黄丸中主要发挥作用的单体之一。Objective：To observe the effects of sera containing Liuwei Dihuang Pill（LDP）,a compound traditional Chinese herbal medicine,and its monomer catalpol on transforming growth factor-β1（TGF-β1）/Smad pathway in HK-2 cells.Methods：Wistar rats were intragastrically administered with LDP twice daily for 3 days and then sera were obtained.HK-2 cells were cultured at different concentrations of serum containing LDP.Lactate dehydrogenase（LDH）activity in the medium and cell activity,tested by methyl thiazolyl tetrazolium（MTT）method,were combined to determine the best concentration of the pill.Hepatocyte growth factor（HGF）was used as a positive control,while Western blotting was applied to observe the effects of sera containing LDP and catalpol on Smad2 phosphorylation and protein expressions of Smad2,Smad7 and SnoN.Results：The 10% concentration of serum containing LDP was selected to carry out this study,as it showed no change of LDH activity in the medium and vitality of cells cultured with serum from normal rats.Similar to the mechanisms of HGF,the 10% concentration of serum containing LDP significantly inhibited the phosphorylation of Smad2 protein and up-regulated the expression of the Smad transcriptional co-repressor SnoN in TGF-β1-stimulated HK-2 cells.Catalpol inhibited phoshporylation of Smad2 without affecting SnoN protein expression.Neither serum containing LDP nor catalpol showed typical regulation effects on Smad7 expression in TGF-β1-stimulated HK-2 cells.Conclusion：Serum containing LDP at 10% concentration has a potent antagonistic action on TGF-β1/Smad pathway in TGF-β1-stimulated HK-2 cells,which is similar to HGF.Catalpol is one of the most important monomers that plays a key role in LDP.

关 键 词：中药 六味地黄丸 中药单体 肝细胞生长因子 HK-2细胞 Smad蛋白类 

分 类 号：R285.5[医药卫生—中药学]