转换生长因子-β1对肺成纤维细胞窖蛋白-1与细胞外基质表达的影响  

Expression of caveolin-I and extracellular matrix induced by transforming growth factor -β1 in human fetal lung fibroblasts

作　　者：丁辉[1] 周凤秋[1] 蔡后荣[1] 周乙华[2] 孟凡青[1] 

机构地区：[1]南京医科大学附属鼓楼临床医学院呼吸内科,210008 [2]南京医科大学附属鼓楼临床医学院 检验科,210008

出　　处：《中华结核和呼吸杂志》2010年第4期280-283,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：基金项目：南京市医学科技发展重点项目资助项目（ZKX08023）

摘　　要：目的研究转换生长因子-β1（TGF—β1）对人胚肺成纤维细胞（HFLF）的窖蛋白一1、I型胶原和d一平滑肌肌动蛋白（α—SMA）表达的影响。方法体外培养HFLF，以未加TGF-β1的HFLF为对照组。分别以不同终浓度（2、5和10μg／L）TGF-β1处理的HFLF为实验组，每组5×10^8细胞，实验重复3次，逆转录一PCR和Westernblot法分别检测TGF-β1刺激3、6、12和24h后各组窖蛋白-1及其mRNA、I型胶原和α—SMA蛋白表达。组间均值比较采用单因素方差分析，采用直线回归模型进行相关性分析。结果TGF-β1作用后，HFLF窖蛋白-1的mRNA和蛋白表达逐渐减少，呈时间一浓度依赖关系；TGF-β1作用12h，中剂量组（5μg／L）窖蛋白一1的mRNA和蛋白表达的积分吸光度值（0．59±0．06和0．53±0．04）明显低于对照组（1．22±0．12和1．45±0．06），差异均有统计学意义（F值分别为29．279和95．786，均P〈0．01）；TGF-β1作用24h，中剂量组（5μg/L）I型胶原和α-SMA蛋白表达的积分吸光度值（1．35±0．09和0．75±0．06）明显高于对照组（0．28±0．04和0．18±O．04），差异均有统计学意义（，值分别为81．221和65．477，均P〈0．01）；相关分析结果显示，HFLF窖蛋白-1的蛋白表达下调与I型胶原和d—SMA蛋白表达上调呈显著负相关（r值分别为-0．923和-0．793，均P〈0．05）。结论TGF-β1可下调HFLF窖蛋白．1的mRNA和蛋白表达，并与I型胶原和Ot—SMA蛋白表达上调呈负相关，提示HFLF窖蛋白-1下调可能与肺间质纤维化的发生和发展有关。Objective To investigate the expressions of caveolin-1, collagen- I and α-SMA in human fetal lung fibroblasts induced by transforming growth factor -β1 （ TGF--β1）- Methods Human fetal lung fibroblasts （HFLF） were cultured in vitro, and exposed to TGF-β1 with different final concentrations （2, 5, 10 μ.g/L）. Cells without TGF-β1 exposure served as the control. Caveolin-1 mRNA and protein, collagen- I and α-SMA proteins in cell （ at least 5 × 10^8 ） lysates, were detected at different points after the treatment of TGF-β1, by RT-PCR, Western blot and Immunohistochemistry. Each experiment was repeated 3 times. One-way analysis of variance （ANOVA） was used to compare data among groups, and linear regression was established for correlation analysis. Results TGF-β1 reduced caveolin-1 mRNA and protein expressions in a dose- and time-dependent manner, measured by integral optical density. Compared with the control group （ 1.22 ±0. 12 and 1.45 ±0.06）, caveolin-1 mRNA and protein expressions （0. 59 ±0. 06 and 0. 53 ± 0.04） were significantly reduced, with statistical significance （ F = 29. 279 and F = 95. 786, P 〈 0.01）, in cells exposed to TGF-β1 （5 μg/L） for 12 h. The collagen-I and ct-SMA protein expressions （ 1.35 ± 0. 09 and 0. 75 ± 0. 06） measured by integral optical density increased after stimulation, peaked at 24 h in 5 μg/L TGF-131 group. At that time point, the collagen- I and α-SMA protein expressions were 4. 2 and 4. 8 times of the control group （0. 28 ± 0. 04 and 0. 18 ± 0. 04）, F = 81. 221 and F = 65.477, P 〈 0. 01. Caveolin-1 expression had a negative eorrelation with collagen- I （ r = - 0. 923, P 〈 0. 05 ） and α-SMA protein expression （ r = - O. 793, P 〈 0. 05 ） . Conclusion The reduced caveolin-1, which was negatively correlated with expressions of collagen- I and α-SMA proteins in HFLF cells exposed to TGF-β1, may be related to the development and progress of pulmonary fibrosis.

关 键 词：成纤维细胞 窖蛋白1 肌动蛋白类 转化生长因子Β1 

分 类 号：R563[医药卫生—呼吸系统]