CSD肽对肺成纤维细胞细胞外基质及Smad信号表达的影响  

Effects of Caveolin-1 Scaffolding Domain Peptide on Expressions of Extracellular Matrix and Smads in Human Fetal Lung Fibroblasts

作　　者：曹敏[1] 丁辉[1] 周乙华[2] 蔡后荣[1] 

机构地区：[1]南京大学医学院附属鼓楼医院呼吸科,江苏南京210008 [2]南京大学医学院附属鼓楼医院科研部,江苏南京210008

出　　处：《中国呼吸与危重监护杂志》2011年第5期442-445,共4页Chinese Journal of Respiratory and Critical Care Medicine

基　　金：南京市医学科技发展重点项目(编号:ZKX08023)

摘　　要：目的研究CSD肽(CSD-p)对转化生长因子β1(TGF-β1)刺激下人胚肺成纤维细胞的细胞外基质及Smad信号表达的影响。方法体外培养人胚肺成纤维细胞,分为4组:对照组(无TGF-β1处理)、TGF-β1处理组、CSD-p处理组、TGF-β1及CSD-p联合处理组。RT-PCR测定各组窖蛋白1mRNA的表达;Western blot检测各组窖蛋白1、α平滑肌肌动蛋白(α-SMA)、胶原Ⅰ、p-Smad2、p-Smad3及Smad7蛋白的表达。结果 TGF-β1刺激人胚肺成纤维细胞后,窖蛋白1的mRNA和蛋白表达较对照组明显降低(mRNA:0.404±0.027比1.540±0.262;蛋白:0.278±0.054比1.279±0.085;P<0.01)。经TGF-β1刺激,人胚肺成纤维细胞的胶原Ⅰ(1.127±0.078比0.234±0.048)、α-SMA(1.028±0.058比0.295±0.024),p-Smad2(1.162±0.049比0.277±0.014)、p-Smad3(1.135±0.057比0.261±0.046)蛋白表达增加(P<0.01);Smad7表达较对照组明显降低(0.379±0.004比1.249±0.046,P<0.001)。与TGF-β1处理组比较,CSD肽明显抑制TGF-β1诱导的胶原Ⅰ(0.384±0.040)、α-SMA(0.471±0.071)、p-Smad2(0.618±0.096)、p-Smad3(0.461±0.057)蛋白表达;上调Smad7的蛋白表达(0.924±0.065比0.379±0.004)。结论 CSD肽下调TGF-β1刺激下人胚肺成纤维细胞的胶原Ⅰ及α-SMA蛋白的表达,可能通过抑制TGF-β1/Smads信号通路激活,提示使用CSD肽增加窖蛋白1功能可能对肺纤维化具有一定的干预作用。Objective To investigate the effects of caveolin-1 scaffolding domain peptide（CSD-p）on expressions of extracellular matrix and Smads in human fetal lung fibroblasts.Methods Human fetal lung fibroblasts were cultured in vitro and divided into four groups.A control group：the cells were cultured in DMEM without TGF-β1 or CSD-p.A CSD-p treatment group：the cells were cultured in DMEM containing 5 μmol/L CSD-p.A TGF-β1 treatment group：the cells were cultured in DMEM containing 5 μg/L TGF-β1.A TGF-β1＋ CSD-p treatment group：the cells were cultured in DMEM containing 5 μg/L TGF-β1 and 5 μmol/L CSD-p.Caveolin-1 mRNA was detected by RT-PCR.Caveolin-1,collagen-Ⅰ,α-SMA,p-Smad2,p-Smad3 and Smad7 proteins were measured by Western blot.Results Compared with the control group,the Caveolin-1 mRNA and protein expressions in the cells of TGF-β1 group significantly reduced（mRNA：0.404±0.027 vs.1.540±0.262;protein：0.278±0.054 vs.1.279±0.085;P0.01）,and the expression levels of collagen-Ⅰ and α-SMA proteins significantly increased（collagen-Ⅰ：1.127±0.078 vs.0.234±0.048;α-SMA：1.028±0.058 vs.0.295±0.024）.Meanwhile,the expression levels of p-Smad2（1.162±0.049 vs.0.277±0.014）and p-Smad3 proteins（1.135±0.057 vs.0.261±0.046）increased with statistical significance（P0.01）,but the expression level of Smad7 protein significantly reduced（0.379±0.004 vs.1.249±0.046,P0.001）.In the CSD-p group,CSD-p had no significant effects on the expressions of above proteins compared with the control group.But in the TGF-β1＋CSD-p group,the overexpressions of collagen-Ⅰ,α-SMA,p-Smad2 and p-Smad3 induced by TGF-β1 were obviously inhibited by CSD-p（collagen-Ⅰ：0.384±0.040 vs.1.127±0.078;α-SMA：0.471±0.071 vs.1.127±0.078;p-Smad2：0.618±0.096 vs.1.162±0.049;p-Smad3：0.461±0.057 vs.1.135±0.057;P0.01）.Otherwise,the up-regulation of Smad7（0.924±0.065 vs.0.379±0.004）was found.Conclusions CSD-p can reduce fibroblast collagen-I and α-SMA protein expressions stimulat

关 键 词：成纤维细胞 窖蛋白1 肺纤维化 转化生长因子Β1 

分 类 号：R563[医药卫生—呼吸系统]