一种小鼠胰岛经门静脉肝内移植方法及疗效评价

A method of islet allotransplantation through portal vein into mouse liver and the effect evaluation

目的介绍一种糖尿病小鼠经门静脉到肝内胰岛移植的方法并进行移植前后的血糖、口服糖耐量、免疫组化等评价。方法采用Balb/c小鼠,将分离纯化的小鼠胰岛培养6 h后,用自制的移植工具进行糖尿病模型小鼠的肝门静脉插管和输注胰岛,移植胰岛量为(320±30)IEQ,对移植前后的小鼠血糖进行测定,在移植后第10天进行糖耐量试验,并取受体小鼠肝脏,进行HE染色和免疫组化分析,观察胰岛细胞团在肝内的存活情况。结果受体小鼠胰岛移植后血糖均能长期维持正常,糖耐量试验结果显示与正常小鼠无统计学差异(P=0.81),组织学结果显示胰岛细胞在小鼠肝内存活良好。结论采用本方法可建立小鼠胰岛经门静脉移植到肝内的模型,为下一步进行胰岛移植相关药物筛选及免疫学等方面研究奠定了基础。

Objective An improved method about how to transplant islets through portal vein into liver in mouse were introduced, and blood glucose, glucose tolerance and histological stain after transplantation were evaluated. Methods Balb/c mice were used in this experiment. The（320±30） IEQ purified isletswere cultured for 6 h before cannulated in portal vein and infused into the liver of mouse by a modified instrument for transplantation.Then the blood glucose was tested regularly, the oral glucose tolerance test was done at the 10 th day and the liver was cut at the 10 th day after the islet transplantation. The HE and immunofluorescence analysis of islet in the liver were also carried out to evaluate the islet morphology and function. Results Mice recipient achieved and maintained normal blood glucose for a long time after islet transplantation. OGTT results at 10 th day after islet transplantation showed that the blood glucose levels of mice had no significant difference with those of normal mice（P=0.81）. And the results of immunohistochemical analysis showed that the activity of islets was good after transplantation. Conclusion The method can establish a model of islets allotransplanted into the mouse liver of mouse via portal vein, which provides a good foundation for further research on drug screening and immunology.