摘要
目的 探讨脂肪特异性蛋白27(Fsp27)基因对活体肝纤维化进程的调节作用.方法 分离原代肝星状细胞(HSCs),采用PCR技术检测在原代HSCs及活化HSCs中的Fsp27基因表达;构建携带Fsp27目的基因的慢病毒;建立肝纤维化模型;慢病毒转染模型鼠肝脏,对实验大鼠行肝脏病理切片检查,采用ELISA法和放射免疫法分别检测肝脏及血浆纤维蛋白含量.结果 Fsp27基因在原代HSCs及活化HSCs中表达差异有统计学意义(P<0.01);成功构建了Fsp27基因的慢病毒载体;成功建立了肝纤维化活体模型;Fsp27基因慢病毒成功转染大鼠肝脏,Fsp27基因减轻了肝脏的纤维化程度.结论 Fsp27基因具有延缓活体肝纤维化进程的作用.
Objective To investigate the influence of fat-specific protein 27 (Fsp27) gene on the regulation of liver fibrogenesis in vivo.Methods Hepatic stellate cells (HSCs) were isolated from rat liver.Fsp27 gene was detected in primary HSCs and activated HSCs by real-time quantitative PCR (RTqPCR).Lentiviral vector carrying Fsp27 gene was constructed.The model of liver fibrosis was established by infusing carbon tetrachloride (CC14).The rats with liver fibrogenesis were infected by the virus.Liver sections were made to observe the structure and form of liver histocytes.The content of fibrous protein in liver and serum was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay.Resukts HSCs were isolated and cultured successfully.The difference of Fsp27 gene between primary HSCs and activated HSCs was significant(P 〈 0.01).The model of liver fibrosis was achieved.After infecting the model rats,we found the fibrosis level in treatment group was lower compared with control group.Conclusions Fsp27 treatment can decrease collagen deposition in the liver and inhibit the formation of fibrosis.
出处
《中国医师杂志》
CAS
2013年第8期1028-1031,共4页
Journal of Chinese Physician
基金
基金项目:温州市科技计划项目(Y20120191)
浙江省重中之重外科学组资助项目(浙教高科2008-255)