摘要
目的:检测转化生长因子β1(TGFβ1)对大鼠肺成纤维细胞(RLF)中转录因子SP1、AP1和Smad3-Smad4活性的影响并探讨其意义。方法:进行大鼠肺成纤维细胞的原代分离培养,应用10μg/L TGFβ1进行处理,通过凝胶阻滞电泳(EMSA)、免疫印迹和免疫组化染色等方法,检测照射后转录因子SP1、AP1和Smad3-Smad4活性的变化及Ⅰ型胶原和Ⅰ型组织纤溶酶原激活物抑制剂(plasminogen activator inhibi-torⅠ,PAI-1)表达的变化。结果:正常RLF中AP1和Smad3-Smad4形成弱阻滞条带,SP1无阻滞条带形成。以10μg/LTGFβ1处理后,3种转录因子所形成的阻滞条带明显增强,PAI-1和Ⅰ型胶原的表达增多。结论:一定浓度的TGFβ1可促进RLF中转录因子SP1、AP1和Smad3-Smad4的活化,并进而增加PAI-1和Ⅰ型胶原的表达。
AIM: To investigate the effects of TGFβ1 on the transcriptive activity of SP1, AP1 and Smad3-Smad4 in rat lung fibroblasts(RLF). METHODS: RLFS were isolated and cultured and then treated with 10 μg/L TGFβ1. The transcriptive activity of SP1, AP1 and Smad3-Smad4 and the expression of plasminogen activator inhibitor I ( PAI-1) and collagen I were detected by means of electrophoretic mobility shift assay ( EMSA), Western blot and immunohistochemical staining. RESULTS: After treatment with 10μg/L TGFβ1, AP1, SP1 and Smad3-Smad4 were activated and the expression of collagen Ⅰ and PAI-1 were increased. CONCLUSION:TGFβ1 may activate SP1, AP1 and Smad3-Smad4 and thus increase the expression of PAI-1 and collagen Ⅰ.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2005年第6期679-682,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(No.30170292)