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

孙静; 李灿; 马燕; 郭莉娜; 田茂岑; 刘琴; 刘卫华; 吴江; 邓峰美; 刘漪沦

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

摘要

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ROSA26 td Tomato（tdTomato）小鼠及Gli1-CreER^t2（Gli1）小鼠进行繁育保种及交配，经PCR基因型鉴定获得Gli1-CreER^t2；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-CreER^t2；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。

Abstract: Objective To establish Gli1-CreER^t2; tdTomato genetic lineage-tracing mice for studies on hepatic fibrosis. Methods Offspring of ROSA26 td Tomato (tdTomato) mice and Gli1-CreER^t2 mice (Gli1 mice) were obtained, with Gli1-CreER^t2; 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-CreER^t2; 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-CreER^t2; 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.

HTML全文

参考文献(12)

施引文献

资源附件(0)

开放获取 本文遵循知识共享署名—非商业性使用4.0国际许可协议（CC BY-NC 4.0），允许第三方对本刊发表的论文自由共享（即在任何媒介以任何形式复制、发行原文）、演绎（即修改、转换或以原文为基础进行创作），必须给出适当的署名，提供指向本文许可协议的链接，同时标明是否对原文作了修改；不得将本文用于商业目的。CC BY-NC 4.0许可协议详情请访问 https://creativecommons.org/licenses/by-nc/4.0