發(fā)布時間：2018-12-14 19:24

【摘要】：目的探討葡萄糖轉(zhuǎn)運蛋白2(Glut2)在人臍帶間充質(zhì)干細胞(h UMSCs)向胰島前體細胞分化過程中的表達變化。方法分離、培養(yǎng)、鑒定及誘導(dǎo)h UMSCs,分別收集誘導(dǎo)過程中第7天、14天和21天細胞和細胞上清液,采用免疫細胞化學(xué)、ELISA、免疫熒光、Western blotting及Real-time PCR檢測誘導(dǎo)后細胞相關(guān)蛋白和基因的表達。結(jié)果免疫組織化學(xué)檢測誘導(dǎo)后細胞胰腺十二指腸同源盒基因-1(PDX-1呈陽性表達;免疫熒光檢測誘導(dǎo)后細胞Ngn3和胰島素呈陽性表達;Western blotting檢測Glut2在誘導(dǎo)過程中逐漸升高,誘導(dǎo)14 d達到峰值,與正常組比較P0.01;Real-time PCR顯示,Glut2基因自第7天即增高(P0.05)。結(jié)論 h UMSCs經(jīng)誘導(dǎo)后分化為胰島前體細胞,表達Glut2后能引起胰島素分泌,已初步具有胰島B細胞功能。
[Abstract]:Objective to investigate the expression of glucose transporter 2 (Glut2) in the differentiation of human umbilical cord mesenchymal stem cells (h UMSCs) into islet precursor cells. Methods the supernatants of cells and cells were collected on the 7th, 14th and 21st day during the induction by UMSCs,. The supernatants were collected by immunocytochemistry and ELISA, immunofluorescence. Western blotting and Real-time PCR were used to detect the expression of cell related proteins and genes. Results the positive expression of PDX-1 was detected by immunohistochemistry, and the expression of Ngn3 and insulin was detected by immunofluorescence. Western blotting detection of Glut2 increased gradually during induction and reached its peak at 14 days after induction. Compared with the normal group, P0.01Real-time PCR showed that the Glut2 gene increased from the 7th day (P0.05). Conclusion h UMSCs can differentiate into islet precursor cells after induction and expression of Glut2 can induce insulin secretion, which has the function of islet B cells.
【作者單位】： 貴州醫(yī)科大學(xué)組織學(xué)胚胎學(xué)教研室;武漢大學(xué)醫(yī)學(xué)研究院;
【基金】：國家自然科學(xué)基金(81160099)
【分類號】：R587.1

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