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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-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。

     

    Abstract: 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.

     

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