血管紧张素Ⅱ对人α1(Ⅰ)胶原基因启动子与活性的影响  

Angiotensin Ⅱ Modulate The Activity Of Promoter From α1(Ⅰ) Procollagen Gene

作　　者：蒋百华[1] 马小琴[2] 徐玲玲[3] 赵文静[3] 

机构地区：[1]浙江省嘉兴市第一医院,浙江嘉兴314000 [2]嘉兴市中医院 [3]第二军医大学长征医院全军免疫中心

出　　处：《中国实验诊断学》2006年第11期1328-1330,共3页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨血管紧张索Ⅱ（AngⅡ）对人皮肤成纤维细胞增殖活性及人α1（Ⅰ）原胶原基因启动调控序列的作用。方法用组织块法原代培养人皮肤成纤维细胞并进行传代培养作为本研究的模型细胞。①采用四甲基偶氮唑盐（MTT）掺入法测定经不同浓度血管紧张素Ⅱ处理24h后成纤维细胞的增殖情况。②构建含长短不一的人α1（Ⅰ）原胶原基因5侧翼区序列与氯霉素乙酰基转移酶（CAT）报告基因的重组体pCOLH1 1．5、pCOLH1 2．5，用FUGENE6将重组体转染至人皮肤成纤维细胞，ELISA法测定不同浓度血管紧张素Ⅱ作用24h后，转染有重组体的成纤维细胞的CAT表达量。结果①在1×10^-9 -1×10^-5mol／L血管紧张素Ⅱ处理24h后，各浓度组成纤维细胞增殖值与对照组之间不存在剂量依赖关系（P〉0．05）。②2种重组体转染成纤维细胞，并经血管紧张素Ⅱ处理24h后，转染细胞的CAT相对表达量测定：pCOLH1 2．5组与对照组之间，较高浓度组（1×100mol／L）与较低浓度组（10×10μmol／L）之间存在显著差异（P〈0.05），且存在剂量依赖关系，pCOLH1 1．5组与对照组之间不存在明显差异（P〉0.05）。结论血管紧张素Ⅱ在1×10^-9-1×10^-5mol／L浓度范围内，对所研究的模型细胞——人皮肤成纤维细胞增殖不存在剂量依赖关系。血管紧张素Ⅱ对胶原基因启动序列pCOLH1 2．5具有正性调控作用，并存在剂量依赖关系。Objective To investigate the effects of Angioteusin Ⅱ on the promoter activities of human α1 （ Ⅰ） procollagen gene.Methods Fibroblasts from human shin were primarily cultured and subcuhmed.① The effects of AngⅡ on human skin fibroblasts were detemrined by MTT incorporation test.② Two plasmids of pCOLH1 1.5 and pCOLH1 2.5 containing various lengths of 5＇ flank sequence of humancd （ Ⅰ ） procollagen gene and CAT as reporter gene were constructed, and were transfected into the human shin fibroblasts by FuGENE traasfection reagent.The effects of AngⅡ on 2 plasminds were determined by CAT-ELISA.Results ①After 24 h of treatment with 1 × 10^-9 - 1 × 10^-5 mol/L Ang Ⅱ in DMEM containing 2% FCS, MMT O .D. values showed no difference among fibroblasts treated with Ang Ⅱ and control （ P 〉 0.05）, ② The 2 plasmids were transfected in fibroblasts and then treated with 10^-5 and 10^-6 mol/L AngⅡ for 24 h,relative CAT value were different between group pCOLH1 2.5 and control,higher AngⅡ （1× 10^-5 mol/L） and lower AngⅡ （1 × 10^-6）（P 〈0.05）. Conclusion AngⅡ has no effect on the proliferation of human skin fibroblasts,and it has positive effect on the promoter activity of pCOLH1 2.5 of human a（ Ⅰ ） procollagen gene,which is dose-depen- dent.

关 键 词：血管紧张素Ⅱ 基因 胶原 启动子 

分 类 号：R33[医药卫生—人体生理学]