糖肾康中药复方含药血清对高糖环境下RMCs的增殖及TGF-β1/Smad2/3信号通路的影响  

Effect of Tangshenkang Granule Containing Serum on Renal Mesangial Cells' Proliferation and TGF-β_1/Smad2/3 Pathway in the High Glucose Condition

作　　者：娄凯[1] 何永[2] 魏静 韩文霞[1] 刘丹丹[4] 宋玉文[5] 姜秀云[1] 于春晓[1] 高聆[6] 管庆波[1] 

机构地区：[1]山东大学附属省立医院内分泌科山东省内分泌代谢病临床医学中心山东省临床医学研究院内分泌与代谢研究所,济南250021 [2]山东中医药大学信息管理学院,济南250355 [3]山东省济南市中医院内分泌科,济南250012 [4]山东省胸科医院,济南250013 [5]山东省立医院西院麻醉科,济南250022 [6]山东大学附属省立医院中心实验室,济南250021

出　　处：《中国中西医结合杂志》2015年第1期88-92,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.30701130);国家自然科学基金资助项目(No.81273880);国家自然科学基金资助项目(No.30971408;81170764);山东省自然科学基金面上项目(No.ZR2009CM101);山东省自然科学基金资助项目(No.Y2008C113);山东省自然科学基金资助项目(No.ZR2009CM101);国家中医药管理局中医临床研究基地业务建设科研专项资助项目(No.JDZX2012007)

摘　　要：目的研究糖肾康中药复方含药血清对高糖环境下大鼠肾系膜细胞(RMCs)增殖及TGF-β1/Smad2/3信号通路的影响。方法 12只SD大鼠采用随机数字法分为小剂量中药组、中剂量中药组、大剂量中药组,空白组,每组3只,灌胃7天后门静脉取血。分别用含中药血清和普通大鼠血清培养RMCs。采用4-甲偶氮唑蓝(MTT)法测定系膜细胞的增殖率,以确定最佳中药血清浓度;实时定量、PCR及酶联免疫吸附剂(ELISA)法测定TGF-β1mRNA及蛋白的表达,蛋白免疫印迹(Western blot)及免疫荧光方法测定Smad2/3蛋白表达及磷酸化水平。结果不同剂量的糖肾康中药含药血清均能抑制大鼠肾小球系膜细胞的增殖;中药血清可显著抑制高糖诱导的TGF-β1表达及Smad2/3磷酸化。结论糖肾康中药含药血清能抑制高糖刺激下的RMCs增殖,其作用机制可能与抑制TGF-β1/Smad2/3通路有关。Objective To study the effect of Tangshenkang Granule（ TG） containing serum on renal mesangial cells’（ RMCs） proliferation and TGF-β1/ Smad2 /3 pathway in the high glucose condition. Methods Twelve SD rats were randomly divided into four groups,i. e.,the low dose TG group,the middle dose TG group,the high dose TG group,and the blank control group,3 in each group. After 7-day gastrogavage via portal vein blood,rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium（ MTT） assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2 /3 protein expression and phosphorylation were determined by We-stern blot and immunofluorescence. Results TG containing serum at different doses could inhibit high glucose induced RMC cells’ proliferation,TGF-β1over-expression and Smad2 /3 phosphorylation. Conclusion TG containing serum could inhibit high glucose induced RMC cells’ proliferation,and its mechanism might be possibly associated with inhibiting TGF-β1/ Smad2 /3 signaling pathway.

关 键 词：糖肾康中药复方 含药血清 TGF-β1/Smad2/3通路 增殖 糖尿病肾病 

分 类 号：R285.5[医药卫生—中药学]