神经胶质瘤相关癌基因锌指蛋白1遗传谱系示踪小鼠在肝纤维化研究中的应用  

Establishing Glioma-associated Oncogene Homolog 1 Genetic Lineage-tracing Mice for Studies on Hepatic Fibrosis

作　　者：孙静[1,2] 李灿 马燕[3] 郭莉娜 田茂岑 刘琴 刘卫华 吴江 邓峰美[1] 刘漪沦[2,3] SUN Jing;LI Can;MA Yan;GUO Li-na;TIAN Mao-cen;LIU Qin;LIU Wei-hua;WU Jiang,;DENG Feng-mei;LIU Yi-lun(Department of Pathophysiology and Pathology,Chengdu Medical College,Chengdu 610500,China;Experimental Center for Medical Science Research,First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China;Department of Surgery,First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China;West China School of Basic Sciences and Forensic Medicine,Sichuan University,Chengdu 610041,China)

机构地区：[1]成都医学院病理学与病理生理学教研室,成都610500 [2]成都医学院第一附属医院科研实验中心,成都610500 [3]成都医学院第一附属医院外科,成都610500 [4]四川大学华西基础医学与法医学院,成都610041

出　　处：《四川大学学报（医学版）》2018年第4期560-565,共6页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金面上项目(No.81570558);四川省科技厅应用基础研究项目(No.2017JY0304;No.2017JY0150);四川省医学会科研课题计划(No.S16021)资助

摘　　要：目的探索神经胶质瘤相关癌基因锌指蛋白1(glioma-associated oncogene homolog 1,Gli1)遗传谱系示踪小鼠在肝纤维化研究中的应用。方法将ROSA26td Tomato(tdTomato)小鼠及Gli1-CreERt2(Gli1)小鼠进行繁育保种及交配,经PCR基因型鉴定获得Gli1-CreERt2;tdTomato目标小鼠。利用CCl4制备肝纤维化小鼠模型。取肝脏组织,制作石蜡切片进行HE和Masson染色。制作冰冻切片,于荧光显微镜下观察红色荧光蛋白tdTomato的表达。结果获得理想数量Gli1-CreERt2;tdTomato目标小鼠。繁殖性能检测发现,亲代与各子代小鼠的繁殖性能无显著性差异(P>0.05)。HE及Masson染色结果发现,CCl4诱导的纤维化模型组假小叶形成。荧光显微镜观察发现,模型组中红色荧光强度明显高于正常对照组。结论 Gli1遗传谱系示踪小鼠可以通过对Gli1阳性细胞的示踪,实现肝纤维化发病过程中纤维组织来源细胞的检测、转化及参与肝纤维的发病机理研究。Objective To establish Gli1-CreERt2;tdTomato genetic lineage-tracing mice for studies on hepatic fibrosis.Methods Offspring of ROSA26 td Tomato（tdTomato）mice and Gli1-CreERt2 mice（Gli1 mice）were obtained,with Gli1-CreERt2;tdTomato genotype being identified by PCR.The mice model of hepatic fibrosis was induced with CCl4.Their liver samples were taken.The formalin-fixed and paraffin-embedded samples were prepared for HE staining and Masson staining.The expression of tdTomato was observed under immunofluorescent microscope.Results An ideal number of Gli1-CreERt2;tdTomato genetic lineage-tracing mice were harvested.The differences in fertility between the parental and the offspring mice were not significant（P〈0.05）.Pseudolobular formation occurred in the CCl4-induced hepatic fibrosis model mice.Enhanced red fluoresce was observed in the model mice.Conclusion Gli1-CreERt2;tdTomato genetic lineage-tracing mice can be used to monitor the cell source of fibrous tissues,its transition as well as the underlying mechanism of pathogenesis of hepatic fibrosis.

关 键 词：遗传谱系示踪 肝纤维化 肌成纤维细胞 

分 类 号：R575.2[医药卫生—消化系统]