论文部分内容阅读
目的探讨联合应用高糖和胰升血糖素样肽-1(GLP-1)、活化素A、尼克酰胺和β细胞调节素(BTC)刺激体外能否诱导人骨髓间充质干细胞向胰岛样细胞分化。方法分别用放射免疫法、RT-PCR和免疫细胞化学方法检测胰岛素浓度、胰岛素mRNA表达以及相关蛋白的表达。结果 (1)刺激后各组可分泌胰岛素量增加。(2)RT-PCR发现GLP-1组、活化素A组、尼克酰胺组、共同刺激组和BTC组等均可产生约300bp的目的片段,即胰岛素原基因的mRNA表达。(3)免疫细胞化学证实除了对照组外,各刺激组均有胰岛素表达;各刺激组均未表达胰升血糖素;除协同作用组有生长抑素的弱阳性表达外,其他各刺激组均未表达生长抑素;尼克酰胺组和协同作用组可见到巢蛋白的弱阳性表达。(4)高糖刺激胰岛素释放实验结果显示,经联合高糖和GLP-1、活化素A、BTC、尼克酰胺和共同刺激诱导分化后,骨髓间充质干细胞(BM-SCs)对高葡萄糖刺激有反应,能相应增加胰岛素的分泌量。结论联合高糖、GLP-1、活化素A、尼克酰胺和BTC等因子可以在体外诱导人BMSCs分化为胰岛素分泌细胞,但胰岛素分泌水平较低。
Objective To investigate whether the combined use of high glucose and GLP-1, activin A, nicotinamide and β-cell stimulating hormone (BTC) can induce human bone marrow mesenchymal stem cells to differentiate into islet-like cells Differentiation. Methods Radioimmunoassay, RT-PCR and immunocytochemical methods were used to detect insulin concentration, insulin mRNA expression and related protein expression. Results (1) After stimulation, the secretion of insulin in each group increased. (2) RT-PCR revealed that the GLP-1 group, activin A group, nicotinamide group, co-stimulation group and BTC group could produce the target fragment of about 300bp, that is, the mRNA expression of proinsulin gene. (3) Immunocytochemistry confirmed that in addition to the control group, each stimulus group had insulin expression; each stimulus group did not express glucagon; in addition to synergistic effect of somatostatin weakly positive expression, the other stimulus group Somatostatin was not expressed; weak positive expression of nestin was observed in nicotinamide group and synergistic group. (4) High glucose stimulation of insulin releaseExperimental results showed that bone marrow mesenchymal stem cells (BM-SCs) induced by high glucose stimulated by combined high glucose and GLP-1, activin A, BTC, nicotinamide and co- A response, a corresponding increase in insulin secretion. Conclusion Combined with high glucose, GLP-1, activin A, nicotinamide and BTC, human BMSCs can be induced to differentiate into insulin-secreting cells in vitro, but insulin secretion is low.