摘要
背景:研究发现胶原三螺旋重复蛋白-1(Cthrc1)可能在肝脏疾病尤其是肝纤维化中发挥重要作用,但对肝星状细胞增殖的影响尚未完全阐明。目的:研究Cthrc1对肝星状细胞增殖的影响。方法:构建Cthrc1重组腺病毒载体,通过尾静脉注射小鼠体内,以实时PCR和蛋白质印迹法分别检测Cthrc1 mRNA和蛋白表达。通过胆总管结扎和3,5-二乙氧基羰基-1,4-二氢-2,4,6-三甲基吡啶(DDC)饲料喂养分别构建小鼠肝纤维化模型,分别尾静脉注射AdCthrc1或Ad-GFP,以免疫荧光法检测肝星状细胞数量。结果:成功构建Cthrc1重组腺病毒载体;实时PCR和蛋白质印迹法证实尾静脉注射Ad-Cthrc1后小鼠肝内Cthrc1 mRNA和蛋白表达均较对照组增高。HE和Masson染色示成功建立了小鼠肝纤维化模型。免疫荧光法显示Cthrc1过表达能明显抑制肝纤维化模型小鼠肝星状细胞的增殖。结论:成功构建了重组腺病毒载体Ad-Cthrc1并可在体内高效稳定表达,能抑制肝星状细胞的增殖,有望成为肝纤维化的治疗靶点。
Background： Collagen triple helix repeat containing protein-1（ Cthrc1） has been reported playing an important role in liver diseases,especially in liver fibrosis,however,its effect on hepatic stellate cells proliferation is not fully clear.Aims： To investigate the effect of Cthrc1 on proliferation of hepatic stellate cells. Methods： Recombinant adenovirus vector of Cthrc1 was constructed. After injecting Ad-Cthrc1 through tail vein,mRNA and protein expressions of Cthrc1 were determined by real-time PCR and Western blotting,respectively. Liver fibrosis model was established by bile duct ligation and fed with 3,5-diethoxycarbonyl-1,4-dihydrocollidine（ DDC） in mice,respectively. The liver fibrosis mice were injected with Ad-Cthrc1 or Ad-GFP through tail vein. Immunofluorescence was used to determine number of hepatic stellate cells. Results： Recombinant Cthrc1-adenovirus vector was successfully constructed. Real-time PCR and Western blotting showed that mRNA and protein expressions of Cthrc1 were increased in Ad-Cthrc1 group than in control group. HE and Masson staining demonstrated that mice model of liver fibrosis was successfully established. Immunofluorescence showed that overexpression of Cthrc1 inhibited significantly the proliferation of hepatic stellate cells. Conclusions： Recombinant adenovirus vector of Ad-Cthrc1 constructed can express stably in vivo,and inhibit the proliferation of hepatic stellate cells.Therefore,Cthrc1 may become a potential target for treatment of liver fibrosis.
出处
《胃肠病学》
2016年第10期592-596,共5页
Chinese Journal of Gastroenterology
基金
江苏省自然科学基金(BK20160420)
南通市科技局南通市社会发展项目(MS22015105)
关键词
胶原三螺旋重复蛋白-1
腺病毒载体
肝星状细胞
细胞增殖
肝硬化
Collagen Triple Helix Repeat Containing Protein-1
Adenovirus Vector
Hepatic Stellate Cells
Cell Proliferation
Liver Cirrhosis
