利拉鲁肽在人骨髓间充质干细胞诱导分化为胰岛素分泌细胞中的作用
Effects of liraglutide on the differentiation of human bone marrow mesenchymal stem cells into insulin producing cells
目的 观察利拉鲁肽在入骨髓间充质干细胞( hBM-MSCs)向胰岛素分泌细胞(IPCs)方向分化诱导中的作用.方法 采用高糖、尼克酰胺和利拉鲁肽3阶段诱导方案对hBM-MSCs进行定向诱导分化,倒置显微镜下观察诱导过程中细胞的形态学变化,双硫腙染色法鉴定诱导后细胞,Western印迹法检测细胞胰腺十二指肠同源盒基因1( PDX-1)、葡萄糖转运蛋白2(GLUT2)、葡萄糖激酶(GK)、胰岛素的蛋白表达,ELISA检测细胞的胰岛素分泌水平.结果 添加10 nmol/L利拉鲁肽作用7d后诱导效率明显增加.诱导过程中细胞形态由长梭形逐渐变为圆形,并聚集生长,至诱导末出现大量圆形葡萄状聚集生长的胰岛样细胞团;双硫腙染色阳性细胞量、细胞PDX-1、GLUT2、GK、胰岛素的蛋白表达、细胞的基础和葡萄糖刺激的胰岛素分泌水平均逐渐增加(均P<0.05).结论 在体外,高糖、尼克酰胺联合利拉鲁肽可使hBM -MSCs定向诱导分化为IPCs.
更多Objective To investigate the effects of liraglutide on the differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) into insulin-producing cells (IPCs).Methods In vitro,hBM-MSCs were induced into IPCs by three-stage induction procedure containing high glucose,nicotinamide,and liraglutide.The morphological change of cells was observed by inverted microscope during induction and the induced cells were confirmed by dithizone(DTZ) staining.The protein expressions of pancreatic and duodenal homeobox-1 (PDX-1),glucose transporter 2 (GLUT2),glucokinase(GK),and insulin in each stage of the induced cells were detected by Western blot.Insulin secretion was measured by ELISA.Results The induced effect was pronounced after adding 10 nmol/L liraglutide for 7 days.Cells began to aggregate and get round gradually during induction,and the morphology of most cells appeared as grape-like aggregation and clustered islet-like cells by the end of induction.The number of DTZ positive cells and the protein expressions of PDX-1,GLUT2,GK,and insulin were increased gradually( P<0.05 ).The basal and glucose-stimulated insulin secretion from induced cells was also increased gradually(P<0.05).Conclusion BM-MSCs could be induced into IPCs by high glucose,nicotinamide,and liraglutide in vitro.
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