摘要
目的 检测肝细胞生长因子 (HGF)及其受体c met在血管外膜 2种表型成纤维细胞中的表达 ,探索其在血管外膜重塑中的作用。方法 用转化生长因子 β1 建立大鼠主动脉外膜成纤维细胞和肌成纤维细胞 2种细胞表型 ,以实时定量逆转录聚合酶链反应技术和蛋白免疫印迹方法分析HGF及c met在 2种不同表型细胞中的表达 ,酶联免疫吸附试验检测 2种细胞无血清培养上清液中HGF的含量。结果 HGFmRNA及HGF蛋白、c metmRNA及c met蛋白在主动脉外膜成纤维细胞、肌成纤维细胞中均有表达 ,在主动脉外膜成纤维细胞转变为肌成纤维细胞后表达明显上调。随转化生长因子 β1 刺激浓度和时间的增加 ,在主动脉外膜成纤维细胞逐渐表达α 平滑肌肌动蛋白转变为肌成纤维细胞的过程中HGF表达逐渐上调。结论 血管外膜成纤维细胞存在局部HGF系统 。
Objective To investigate whether adventitia is a source of hepatocyte growth factor (HGF) and its receptor c met or not Methods Myofibroblasts (MF) were established by the treatment of adventitial fibroblasts (AF) with transforming growth factor β 1 Expression of HGFmRNA and c metmRNA in two types of cells were evaluated by Real time Quantitative Reverse Transcription Polymerase Reaction The protein level of HGF and c met were detected by western blotting and the content of HGF in the condition medium of the two cells were determined by Enzyme linked immunoabsorbent assay Results The expression of HGF and c met could be detected in both AF and MF either in the ribonucleinic acid level or in the protein level and the expression levels of them in MF were both significantly higher than those of in AF During the course of the phenotypic transformation of MF from AF identified by the expression α SM actin, the expression of HGF was gradually upregulated by transforming growth factor β 1 in a dose dependent and time dependent manner Conclusion Local HGF system really exists in adventitia It is suggested that HGF might play a role in the phenotypic transformation of AF
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2003年第2期132-135,共4页
Chinese Journal of Cardiology
基金
国家重点基础研究"973"(G2 0 0 0 0 56 90 0 4 )
国家自然科学基金 ( 30 0 70 30 3)