利用Cre-Loxp技术构建标记肝星状细胞的小鼠模型  

Constructing transgenic mice to label hepatic stellate cells by Cre-Loxp technology

作　　者：黄紫薇 赵殿元 徐龙 唐丽 Huang Ziwei;Zhao Dianyuan;Xu Long;Tang Li(Dept of Immunology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230032;Institute of Lifeomics,Academy of Military Medical Sciences,Academy of Military Sciences,Bejing 102206;State Key Laboratory of Proteomics,Bejing Proteome Research Center,National Center for Protein Sciences,Beijing 102206)

机构地区：[1]安徽医科大学基础医学院免疫教研室,合肥230032 [2]军事科学院军事医学研究院生命组学研究所,北京102206 [3]蛋白质组学国家重点实验室,北京蛋白质组研究中心,国家蛋白质科学中心,北京102206

出　　处：《安徽医科大学学报》2023年第7期1065-1070,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金杰出青年基金项目(编号:82225009);国家自然科学基金面上项目(编号:32270941)。

摘　　要：目的通过Cre-Loxp技术构建转基因小鼠,应用Lrat^(Cre)工具鼠示踪肝星状细胞分布。方法将Lrat^(Cre)小鼠与H11^(LoxP-ZsGreen-Stop-LoxP-tdTomato)报告基因小鼠交配,通过PCR鉴定得Lrat^(Cre)H11^(LoxP-ZsGreen-Stop-LoxP-tdTomato)报告基因小鼠。固定小鼠肝脏切片,借助免疫荧光标记肝脏非实质细胞抗体,通过Imaris 3D还原分析肝星状细胞和其他肝脏非实质细胞间的表达与分布,以及进行肝星状细胞的自定位检验。结果Lrat^(Cre)介导的红色荧光蛋白tdTomato的表达可以特异性标记肝星状细胞,并通过肝星状细胞与肝脏巨噬细胞、肝窦内皮细胞的免疫荧光定位为细胞间相互作用分析提供了途径。结论Lrat^(Cre)可以介导tdTomato对肝星状细胞进行示踪,同时可用做肝星状细胞特异性的报告基因工具鼠。Objective To trace and investigate the distribution of hepatic stellate cells by Cre-Loxp technology of Lrat^(Cre)mice.Methods Lrat^(Cre)mice were mated with H11^(LoxP-ZsGreen-Stop-LoxP-tdTomato)reporter mice and genotype of their progeny was identified by PCR.The liver sections were fixed,the antibody of liver non-parenchymal cells was labeled with immunofluorescence,and the expression and distribution of liver stellate cells and other liver non-parenchymal cells were analyzed by Imaris 3D reduction,as well as positively identify dual positioning of hepatic stellate cells.Results Lrat^(Cre)H11^(LoxP-ZsGreen-Stop-LoxP-tdTomato)reporter mice specificiallyled to expression of red fluorescent protein tdTomato in hepatic stellate cells,providing a way to analyze the interactions between hepatic stellate cells,Kupffer cells and liver sinusoidal endothelial cells by immunofluorescence localization.Conclusion Lrat^(Cre)strain as a specific reporter mouse for hepatic stellate cells can be uesd to trace hepatic stellate cells.

关 键 词：肝星状细胞 肝脏巨噬细胞 肝窦内皮细胞 报告基因小鼠 CRE-LOXP 

分 类 号：R392-33[医药卫生—免疫学] R329.2[医药卫生—基础医学]