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促肝细胞生长素对大鼠肝星状细胞增殖及α1(Ⅰ)胶原基因启动子活性的影响

Influence of hepatocyte growth-promoting factor on proliferation of hepatic stellate cells and activities of α1(Ⅰ ) procollagen gene promoter in rats

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【作者】 周长林高春芳朱樑宋维林继军袁志忠张忠兵

【Author】 ZHOU Chang-Lin,GAO Chun-Fang, ZHU Liang,SONG Wei, LI Ji-Jun, YUAN Zhi-Zhong1, ZHANG Zhong-Bing (1. Department of VIP Patient, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China; 2. Department of Gastroenterology .Changzheng Hospital; 3. Department of Laboratory Diagnosis,Changzheng Hospital)

【机构】 第二军医大学长征医院贵宾诊疗科长征医院实验诊断科长征医院消化内科长征医院消化内科 上海 200003 长征医院消化内科上海 200003上海 200003

【摘要】 目的:探讨促肝细胞生长素(pHGF)对大鼠肝星状细胞HSC-T6增殖及α1(Ⅰ)胶原基因启动子活性的影响。方法:(1)采用MTT法分析pHGF对HSC-T6细胞增殖的影响。(2)构建含人α1(Ⅰ)胶原基因5′侧翼序列(启动子)与氯霉素乙酰基转移酶(CAT)报告基因的重组体 pCOLH1.5,以脂质体法转染HSC-T6细胞,ELISA法测定不同浓度pHGF作用24 h后,转染了重组体质粒的HSC-T6细胞的CAT表达量。结果:(1)不同浓度pHGF对HSC-T6细胞增殖均有明显的抑制作用,与对照组比较有明显差异(P<0.01)。(2)pHGF在200 μg/ml时对转染重组体质粒pCOLH1.5的HSC-T6细胞的CAT活性具有明显的抑制作用,与对照组相比差异显著(P<0.05);增加pHGF浓度至400μg/ml时,对CAT活性抑制更为明显,与对照组相比差异非常显著(P<0.01)。结论:pHGF对HSC-T6细胞增殖具有明显的抑制作用;对α1(Ⅰ)胶原基因启动子活性具有负性调控作用。

【Abstract】 Objective: To study the effect of hepatocyte growth-promoting (pHGF) on the proliferation of hepatic stellate cell-T6(HSC-T6)and the activities of α1( Ⅰ) procollagen gene promotor. Methods; (1) Proliferation of HSC-T6 was analyzed by mitochondrial reduction of tetrazolium salt (MTT). (2)The recombinant plasmid pCOLH 1. 5 containing 5’ flank sequence of human α1( Ⅰ ) procollagen gene and chloramphenicol acetyltransf erase (CAT) was transfected into HSC-T6. The activities of CAT were assessed by ELISA method 24 h after pHGF treating. Results: (1) pHGF of different concentrations had inhibiting effects on the proliferation of HSC-T5 vs control group (P<0. 01). (2) pHGF at 200 jug/ml markedly inhibited the activities of CAT in HSC-T6 after transfection of pCOLH1. 5 vs control group (P<0. 05), and when pHGF at 400 μg/ml,the inhibiting effect was more obvious(P<0. 01). Conclusion: pHGF has inhibiting effects on the proliferation of HSC-T6 and negative effects on the activities of α1( Ⅰ ) procollagen gene promotor.

【基金】 国家自然科学基金(19872074);上海市科技攻关重点项目(011209014)
  • 【文献出处】 第二军医大学学报 ,Academic Journal of Second Military Medical University , 编辑部邮箱 ,2003年12期
  • 【分类号】R575.2
  • 【被引频次】4
  • 【下载频次】68
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