一種小鼠胰島經門靜脈肝內移植方法及療效評價

蔣煊，傅紅興，邱凱燕，何云強，徐艷艷，祝青卉（.溫州醫科大學　藥學院，浙江　溫州　35035；.麗水市中心醫院　藥劑科，浙江　麗水　33000）

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一種小鼠胰島經門靜脈肝內移植方法及療效評價

蔣煊1，傅紅興1，邱凱燕1，何云強1，徐艷艷2，祝青卉1
（1.溫州醫科大學藥學院，浙江溫州325035；2.麗水市中心醫院藥劑科，浙江麗水313000）

[摘 要]目的 介紹一種糖尿病小鼠經門靜脈到肝內胰島移植的方法并進行移植前后的血糖、口服糖耐量、免疫組化等評價。方法 采用Balb/c小鼠，將分離純化的小鼠胰島培養6 h后，用自制的移植工具進行糖尿病模型小鼠的肝門靜脈插管和輸注胰島，移植胰島量為（320±30）IEQ，對移植前后的小鼠血糖進行測定，在移植后第10天進行糖耐量試驗，并取受體小鼠肝臟，進行HE染色和免疫組化分析，觀察胰島細胞團在肝內的存活情況。結果 受體小鼠胰島移植后血糖均能長期維持正常，糖耐量試驗結果顯示與正常小鼠無統計學差異（P=0.81），組織學結果顯示胰島細胞在小鼠肝內存活良好。結論 采用本方法可建立小鼠胰島經門靜脈移植到肝內的模型，為下一步進行胰島移植相關藥物篩選及免疫學等方面研究奠定了基礎。

[關鍵詞]胰島移植；門靜脈；療效；Balb/c小鼠

大學生創新訓練項目（201410343002）。

A method of islet allotransplantation through portal vein into mouse liver and the effect evaluation JIANG Xuan1,

FU Hong-xing1,QIU Kai-yan1,HE Yun-qiang1,XU Yan-yan2,ZHU Qing-hui1.1School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou,Zhejiang 325035,China;2Department of Pharmacy,Lishui Central Hospital,Lishui,Zhejiang 313000,China

Abstract objectiveAn 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.MethodsBalb/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 10thday and the liver was cut at the 10thday 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.ResultsMice recipient achieved and maintained normal blood glucose for a long time after islet transplantation.OGTT results at 10thday 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.ConclusionThe 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.

Key words islet transplantation; portal vein; curative effect; Balb/c mouse

胰島經肝門靜脈輸注使細胞定植于肝臟是臨床胰島移植常用的方法，并在相關疾病治療中顯示了良好的效果[1-3]。小鼠胰島經肝門靜脈移植治療小鼠糖尿病的模型建立難度較大，該手術需要成熟的小鼠胰島分離技術、培養和活性鑒定技術、顯微外科操作和術后護理經驗，小鼠門靜脈插管出血風險大，因此國內外鮮有文獻報道。……