检验医学与临床
檢驗醫學與臨床
검험의학여림상
JOURNAL OF LABORATORY MEDICINE AND CLINICAL SCIENCES
2015年
8期
1070-1073
,共4页
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
膠質纖維痠性蛋白啟動子%增彊綠色熒光蛋白%肝星狀細胞%肝纖維化
효질섬유산성단백계동자%증강록색형광단백%간성상세포%간섬유화
glial fibrillary acidic protein promoter%enhanced green fluorescent protein%hepatic stellate cells%hepatic fibrosis
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(DESM IN )和α‐平滑肌肌动蛋白(α‐SM A )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。
目的:探索體內特異性靶嚮肝星狀細胞(HSC)治療肝纖維化的技術方法。方法建立四氯化碳誘導的小鼠肝纖維化模型,構建膠質纖維痠性蛋白(GFAP)啟動子調控增彊綠色熒光蛋白(EGFP)錶達載體,採用尾靜脈高壓註射方法將EGFP錶達裸質粒轉染到纖維化小鼠的肝髒。熒光顯微鏡觀察EGFP在肝髒的錶達及免疫熒光共定位,觀測 EGFP 與 HSC 標誌蛋白(DESM IN )和α‐平滑肌肌動蛋白(α‐SM A )的共定位。結果 EGFP 與DESMIN和α‐SMA共定位說明EGFP主要在HSC中錶達,GFAP啟動子能夠調控外源基因在體內靶嚮 HSC。結論 GFAP啟動子介導體內靶嚮 HSC ,治療肝纖維化策略具有臨床應用的潛力。
목적:탐색체내특이성파향간성상세포(HSC)치료간섬유화적기술방법。방법건립사록화탄유도적소서간섬유화모형,구건효질섬유산성단백(GFAP)계동자조공증강록색형광단백(EGFP)표체재체,채용미정맥고압주사방법장EGFP표체라질립전염도섬유화소서적간장。형광현미경관찰EGFP재간장적표체급면역형광공정위,관측 EGFP 여 HSC 표지단백(DESM IN )화α‐평활기기동단백(α‐SM A )적공정위。결과 EGFP 여DESMIN화α‐SMA공정위설명EGFP주요재HSC중표체,GFAP계동자능구조공외원기인재체내파향 HSC。결론 GFAP계동자개도체내파향 HSC ,치료간섬유화책략구유림상응용적잠력。
Objective To find the possibility that the glial fibrillary acidic protein (GFAP) promoters modu‐late 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 in‐jected into mice liver by hydrodynamics based transfection method .EGFP expression in the liver was observated un‐der Fluorescence microscopy to evaluate the efficiency of GFAP promoter .The co‐expressions of EGFP with DESMIN and α‐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 and α‐SMA .Illustrated that GFAP promoter could drive EGFP specific expression in HSCS but not in hepatocytes of liver fibrosis murine .Conclusion These 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 .