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Fah基因剔除小鼠在肝细胞移植中的应用及其病理学变化

Application of Fah knockout mice in the hepatocyte transplantation and the pathological changes

  • 摘要: 目的 : 观察外源成体肝细胞在Fah-/-小鼠肝脏中增殖的微观特性并对该小鼠模型肝脏损伤后的病理学变化进行研究。 方法 : 野生型小鼠成体肝细胞经脾脏移植到Fah-/-小鼠后,观察移植Fah-/-小鼠的体重变化,生存状态及肝组织的Fah免疫组化和电镜观察以评价再殖效率和病理学变化。 结果 : Fah-/-小鼠的肝HE染色显示为亚急性损伤的病理特点。电镜观察显示,初期肝细胞坏死,凋亡现象随时间增多,随后有凋亡和坏死并存。而Fah-/-小鼠在NTBC[2-(2-硝基-4-三氟甲基苯甲酰基)-环己烷-1,3-二酮]药物的治疗后,无明显病理学变化。肝细胞移植后的Fah-/-小鼠均健康存活,体重稳步上升。8周时可以达到90%以上再殖。肝脏生化指标恢复正常。 结论 : Fah-/-小鼠的肝脏损伤可以使移植的肝细胞90%以上的再殖,维持肝脏正常的组织结构,并发挥正常的肝细胞功能。Fah-/-小鼠作为肝脏再殖模型可用于干细胞肝向分化研究。

     

    Abstract: Aim :To investigate the pathological changes in NTBC[2-(2-nitro-4-trifluoro-methyl-benzoyl)-1,3 cyclohexanedione]-induced hepatic injury in mice and in the repopulation of adult hepatocytes in Fah-/- mouse. Methods :Autogenous hepatic injuries in Fah-/- mice were induced by the treatment of NTBC.Injection of hepatocytes obtained from wild-type mice to spleen were transplanted into the Fah-/- mice.Then,changes to body weight and the likelihood of the transplanted Fah-/- mice,and hepatic immunohistochemistry were observed.In addition,pathological changes to liver damage induced by NTBC treatment were analyzed under HE-staining microscopy and electron microscopy. Results :The surviving Fah-/- mice subjected to hepatocyte transplantation were found to be healthy and in stable body weight.Liver repopulation reached to 90% in the 8th week.Repopulating hepatocytes caused no alteration to histological structure of the recipient liver,and subacute hepatic injury occurred in the Fah-/- mice after NTBC treatment.Electronic microscopy observations indicated that necrosis in the hepatocytes occurred at early stage and that apoptosis gradually appeared.It was also shown both necrosis and apoptosis co-existed in the same samples of interest at the following stages of the induced liver injury. Conclusion :Transplanted hepatocytes proliferated in Fah-/- mice allow 90% of the hepatocytic repopulation.Repopulation renders normal hepatic function and structure in the recipient.Fah-/- mice,as a model of liver repopulation,could be applicable in study of stem cell derived hepatic cells in transplantation assay.

     

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