GFAP启动子介导EGFP靶向肝星状细胞治疗肝纤维化的价值探索  

The value of treatment to hepatic fibrosis with GFAP promoter directs EGFP targeting to hepatic stellate cell

作　　者：范建华[1] 徐曼妮[2] 

机构地区：[1]上海市黄浦区中心医院消化内科,200002 [2]同济大学医学院预防医学教研室,上海200092

出　　处：《检验医学与临床》2015年第8期1070-1073,共4页Laboratory Medicine and Clinic

摘　　要：目的探索体内特异性靶向肝星状细胞(HSC)治疗肝纤维化的技术方法。方法建立四氯化碳诱导的小鼠肝纤维化模型,构建胶质纤维酸性蛋白(GFAP)启动子调控增强绿色荧光蛋白(EGFP)表达载体,采用尾静脉高压注射方法将EGFP表达裸质粒转染到纤维化小鼠的肝脏。荧光显微镜观察EGFP在肝脏的表达及免疫荧光共定位,观测EGFP与HSC标志蛋白(DESMIN)和α-平滑肌肌动蛋白(α-SMA)的共定位。结果 EGFP与DESMIN和α-SMA共定位说明EGFP主要在HSC中表达,GFAP启动子能够调控外源基因在体内靶向HSC。结论 GFAP启动子介导体内靶向HSC,治疗肝纤维化策略具有临床应用的潜力。Objective To find the possibility that the glial fibrillary acidic protein（GFAP）promoters modulate the exogenous gene specific expression in HCS for hepatic fibrosis treatment.Methods Established a model of carbon tetrachloride-induced hepatic fibrosis in mice.pCDNA3.1-mGFAP-p-EGFP plasmid was constructed and injected into mice liver by hydrodynamics based transfection method.EGFP expression in the liver was observated under Fluorescence microscopy to evaluate the efficiency of GFAP promoter.The co-expressions of EGFP with DESMIN andα-SMA was detected by immunofluorescence.Results The expression of EGFP in liver driven by GFAP promoter was co-localized with HCS specific markers such as DESMIN andα-SMA.Illustrated that GFAP promoter could drive EGFP specific expression in HSCS but not in hepatocytes of liver fibrosis murine.ConclusionThese observation demonstrate that GFAP promoter could in vivo regulate expression of target gene in HSC,offering a useful tool for gene therapy against hepatic fibrosis.

关 键 词：胶质纤维酸性蛋白启动子 增强绿色荧光蛋白 肝星状细胞 肝纤维化 

分 类 号：R975[医药卫生—药品]