肝细胞生长因子对系膜细胞增殖及TGF-β1启动子活性的影响  

Regulation of mesangial cell proliferation and promotor activity from TGF-β1 by HGF

作　　者：周春华[1] 于海[1] 高春芳[2] 包广宇[2] 

机构地区：[1]海军总医院肾内科,北京100037 [2]第二军医大学长征医院实验诊断科,上海200003

出　　处：《中国现代医学杂志》2006年第22期3409-3411,3414,共4页China Journal of Modern Medicine

摘　　要：目的观察肝细胞生长因子(HGF)对大鼠系膜细胞增殖及对TGF-β1启动子活性的作用。方法应用MTT法观察HGF(0、0.1、1.0、1.0和100.0ng/mL)对大鼠系膜细胞增殖的影响;构建含不同长度人TGF-β1基因启动子片段的重组体phTGF2.14、phTGF1.12,以氯霉素乙酰基转移酶(CAT)为报告基因,并利用脂质体法将其转染至大鼠系膜细胞中,ELISA方法检测报告基因CAT的活性,以观察不同浓度的HGF(1.0、10.0ng/mL)对TGF-β1启动子活性的作用。结果大鼠系膜细胞经不同浓度的HGF干预后,各实验组与对照组相比细胞增殖水平差异无显著性,P>0.05。在系膜细胞中,HGF的浓度分别为1.0和10.0ng/mL时对重组体phTGF2.14及phTGF1.12的表达活性无抑制作用。结论HGF不能直接抑制大鼠系膜细胞增殖。在系膜细胞中HGF对TGF-β1启动子的表达活性无作用,提示其拮抗TGF-β1生物学作用的靶点并不是发生在基因转录水平。[Objective] To study the effect of HGF on the proliferation of rat mesangial cell（RMC） and expressive activity of TGF-β1 gene promoter. [Methods] Proliferation of RMC was analyzed by mitochondrial reduction of tetrazolium salt （MTT）. Two plasmids containing various lengths of 5＇flank sequence of human TGF-β1 gene and CAT as reporter gene were constructed, and were transfected into RMC by FuGENE transfeetion reagent. The effects of HGF（1.0 ng/mL, 10.0 ng/mL）on two plasmids were determmed by CAT-ELISA. [Results] The proliferation of RMC were interfered with HGF of 0, 0.1, 1.0, 10.0, 100.0 ng/mL. There were no significant difference between cells treated with different concentration of HGF and control （P 〉0.05）. The RMC transfected with phTGF＇2.14 or phTGF1. 12 were treated with 1.0 ng/mL and 10 ng/mL. There were no significant difference between test groups and control（P 〉0.05）. [Conclusion] HGF can not affect the proliferation of RMC. HGF can not inhibit the activity of TGF-β1 gene promoter.

关 键 词：肝细胞生长因子 TGF—betal 启动子 

分 类 号：Q784[生物学—分子生物学]