葡萄糖转运蛋白2在人脐带间充质干细胞向胰岛前体细胞分化过程中表达的变化

doi:10.16098/j.issn.0529-1356.2017.04.007

摘要/Abstract

摘要：

目的探讨葡萄糖转运蛋白2（Glut2）在人脐带间充质干细胞(hUMSCs)向胰岛前体细胞分化过程中的表达变化。方法分离、培养、鉴定及诱导hUMSCs，分别收集诱导过程中第7天、14天和21天细胞和细胞上清液，采用免疫细胞化学、ELISA、免疫荧光、Western blotting及Real-time PCR检测诱导后细胞相关蛋白和基因的表达。结果免疫组织化学检测诱导后细胞胰腺十二指肠同源盒基因-1（PDX-1呈阳性表达；免疫荧光检测诱导后细胞Ngn3和胰岛素呈阳性表达；Western blotting检测Glut2在诱导过程中逐渐升高，诱导14 d达到峰值，与正常组比较P<0.01；Real-time PCR显示，Glut2基因自第7天即增高（P<0.05）。结论 hUMSCs经诱导后分化为胰岛前体细胞，表达Glut2后能引起胰岛素分泌，已初步具有胰岛B细胞功能。

关键词: 人脐带间充质干细胞, 胰岛前体细胞, 葡萄糖转运蛋白2, 免疫组织化学, 免疫荧光, 实时定量聚合酶链反应

Abstract:

Objective To detect the expression of glucose transperter 2 (Glut2) when human umbilical cord mesenchymal stem cells (hUMSCs) were transformed into islet precursor cells. Methods hUMSCs were isolated, cultivated, identified and differentiated. The cells and supernatant were collected on the 7th,14th and 21st day of the induction. Immunocytochemistry，immunofluorescence, ELISA, Western blotting and Realtime PCR were used to detect the expression of cell-associated proteins and genes. Results 1.The expression of pancreas duodenum homeobox-1 (PDX-1) was positive; 2.Positive expressions of Ngn3 and insulin were detected by immunofluorescence; 3.The expression of Glut2 protein was measured by Western blotting. The Glut2 protein gradually increased in the induction process, and reached the peak value on the 14th day, compared with the normal group (P<0.01). Glut2 gene increased from the 7th day（P<0.05）in Real-time PCR. Conclusion After induction, hUMSCs turn into pancreatic precursor cells and have functional features of islet B cell initially for expressing Glut2 protein, which can cause insulin secretion.

Key words: Human umbilical cord mesenchymal stem cell, Islet precursor cell, Glucose transporters 2, Immunohistochemistry, Immunofluorescence, Real-time PCR

覃晓莉檀梦天洪艳. 葡萄糖转运蛋白2在人脐带间充质干细胞向胰岛前体细胞分化过程中表达的变化[J]. 解剖学报, 2017, 48(4): 410-415.

QIN Xiao-li TAN Meng-tian HONG Yan. Expression of glucose transporter 2 during differentiation of human umbilical cord mesenchymal stem cells into islet precursor cells [J]. Acta Anatomica Sinica, 2017, 48(4): 410-415.

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