转化生长因子-β_1/Smad信号通路在肾小球系膜细胞的表达及血管紧张素转化酶抑制剂对其影响  

Expression of Transforming Growth Factor-β_1/Smad Signaling Pathway in Rat Glomerular Mesangial Cell and Interventional Effects of Angiotensin-Converting Enzyme Inhibitor

作　　者：邓颖[1] 于力[1] 陈蓉燕[1] 温跃强[1] 温捷[1] 郝志宏[1] 

机构地区：[1]广州医学院附属广州市第一人民医院儿科,广东广州510180

出　　处：《中华妇幼临床医学杂志（电子版）》2011年第4期243-247,共5页Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition)

基　　金：广州市卫生局科研基金项目(2009-YB-028);广东省医学科研基金项目(A2010450)~~

摘　　要：目的探讨转化生长因子(transforming growth factor,TGF)-β1/Smad信号传导通路在肾小球系膜细胞(glomerular measanial cell,GMC)中的作用,观察血管紧张素转化酶抑制剂(angiotensin-converting enzyme inhibitor,ACEI)福辛普利(fosinopril,Fos)在该通路中对磷酸化Smad2/3(phosphorylationSmad2/3,P-Smad2/3)和Ⅰ型胶原蛋白(collagenⅠ,ColⅠ)表达的影响,阐明福辛普利的肾保护作用机制。方法采用经典方法进行大鼠肾小球系膜细胞复苏、培养和传代后,将其按处理方式分为3组:TGF-β1组(TGF-β15ng/mL处理);②Fos组(TGF-β15ng/mL+Fos1×10-4mol/L处理);③对照组(空白)。甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)比色法分别检测3组肾小球系膜细胞增殖光密度(optical density,OD)值,计算细胞抑制率;间接免疫荧光法检测磷酸化Smad2/3表达情况;酶联免疫吸附测定(enzyme linkedi mmunosorbent assay,ELISA)法测定Ⅰ型胶原蛋白的变化,计算抑制率。结果各时间点肾小球系膜细胞增殖荧光强度比较,TGF-β1组明显高于对照组,Fos组低于TGF-β1组,差异有显著意义(P<0.05)。磷酸化Smad2/3在对照组仅极微量表达,TGF-β1组在刺激30min后,荧光强度显著增强,且集中于细胞核。与TGF-β1组相比,Fos组磷酸化Smad2/3表达下降,细胞荧光强度较弱,差异有显著意义(P<0.05)。对照组Ⅰ型胶原蛋白仅少量基础性分泌。与对照组相比,TGF-β1组在各时间点Ⅰ型胶原蛋白分泌量显著上升,差异有显著意义(P<0.05);与TGF-β1组比较,Fos组Ⅰ型胶原蛋白在15min和6h的表达量比较,差异无显著意义(P>0.05);在12h和24h比较,差异有显著意义(P<0.05)。结论人重组转化生长因子-β1能激活体外培养的大鼠肾小球系膜细胞磷酸化-Smads表达,并激活转化生长因子-β/Smads信号途径,该途径激活能致Ⅰ型胶原分泌增多,并能诱导肾小球系膜细胞增殖,其作用呈一定的时间依赖性关系。福辛普利能部分阻断转化生长因子-β/Objective To investigate the role of transforming growth factor （TGF）-β1/Smad signaling pathway in glomerular mesangial cell （GMC）, and the influence of angiotensin-converting enzyme inhibitor （ACEI） fosinopril （Fos） on the expression of phosphorylation-Smad2/3 （P-Smad2/3） and collagen I （Col I ） in this way. Methods Glomerular mesangial ceils of rat in vitro were cultured, and they were divided into 3 groups： OTGF-β1 group （TGF-131 5 ng/mL） ; ②Fos group （TGF-β1; 5 ng/mL＋Fos 1 X 10^-4 mol/L）; @control group （blank）. Methyl thiazolyl tetrazolium （MTT） colorimetric method detected cellular proliferation. The change of phosphorylation-Smad2/3 expression was detected by indirect immunofluorescence. The expression of collagen I in cell culture supernatant was detected by the enzyme- linked immunosorbent assay （ELISA）. Results The optical density（OD） value of cellular proliferation was higher than that in TGF-131 group when compared with control group（P〈0.05）, while lower in Fos group when compared with TGFq3a group （P〈0. 05）. Phosphorylation-Smad2/3 slightly expressed in control group, while increased obviously in TGF-β1; group compared with control group（P〈0.05）. In Fos group, the expression of phosphorylation-Smad2/3 were decreased apparently at 12 h and 24 h than those in TGF-β group（ P 〈0. 05 ）. Conclusion TGF-β1 can activate the expression of phosphorylation-Smads and TGF-β/Smad signaling pathway in glomerular mesangial cell, which will cause the increased expression of collagen I and proliferation of glomerular mesangial cell. Fos can Dartiallv block the activation oftransforming growth factor-β1/Smad signaling pathway in glomerular mesangial cell, inhibit the excretion of collagen I and proliferation of glomerular mesangial cell.

关 键 词：肾小球系膜细胞 Ⅰ型胶原蛋白 转化生长因子-Β1 SMAD蛋白 血管紧张素转化酶抑制剂 

分 类 号：R692[医药卫生—泌尿科学]