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目的探讨血管紧张素Ⅱ(AngⅡ)对人α1(Ⅰ)胶原基因启动子活性的调控作用和川芎嗪干预的影响环节。方法将2.5kb长度的人α1(I)胶原基因启动子片段与氯霉素乙酰基转移酶(CAT)“报告基因”组成的重组体pCOLH2.5,采用脂质体法转染至培养的大鼠心肌成纤维细胞,分别加入AngⅡ和川芎嗪,ELISA法测定CAT活性。结果AngⅡ可明显促进pCOLH2.5片段的启动活性,川芎嗪能抑制AngⅡ的促pCOLH2.5启动活性(P<0.05)。结论川芎嗪能在转录水平上阻断AngⅡ诱导的人α1(I)胶原基因启动子活性增加,从而抑制I型胶原的合成,阻滞和延缓心肌纤维化的发生发展。
Objective To investigate the regulation of angiotensin II (Ang II) on the promoter activity of human α 1 (I) collagen gene and the influence of tetramethylpyrazine on its regulation. Methods The recombinant pCOLH2.5 consisting of a 2.5 kb-length human α1(I) collagen gene promoter fragment and a chloramphenicol acetyltransferase (CAT) reporter gene was transfected into large cultures by liposome method. Rat cardiac fibroblasts were treated with AngII and Tetramethylpyrazine, and CAT activity was measured by ELISA. Results AngII significantly promoted the activation of pCOLH2.5. Ligustrazine could inhibit the activation of pCOLH2.5 by AngII (P<0.05). Conclusion Ligustrazine can block the increase of AngII-induced human α1(I) collagen gene promoter activity at the transcriptional level, inhibit the synthesis of type I collagen, block and delay the occurrence and development of myocardial fibrosis.