摘要
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 Observe the hBMSCs differentiation into insulin-producing cells by stimulation of high glucose, GLP-1, activin A, nicotinamide and BTC respectively. Methods Using the methods of radioimmunoassay, RT-PCR and immunocytochemistry, we deteced the insulin level, insulin mRNA and related proteins. Results 1. The insulin secretory amount of the hBMSCs differentiated cells was increased. 2. The groups of GLP-1, aetivin A, nicotinamide and BTC could express specific proinsulin mRNA fragments at 300bp. 3. Immunocytochemistry confirmed that all stimulating groups showed the positive insulin expression, except for the common L-DMEM group. Glucagon was negative in all stimulating groups. Except for combined treatment group, somatostatin was negative in all stimulating groups. Only nicotinamide group and combined treatment group could express nestin weakly. 4. The results of high glucose stimulated insulin release test showed that the insulin content was increased in all stimulating groups. Conclusions High glucose, GLP-1, activin A, nicotinamide and BTC could induce hBMSCs into insulin producing cells in vitro.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2009年第7期505-509,共5页
Chinese Journal of Diabetes
