地黄多糖诱导骨髓间充质干细胞为神经样细胞后的突触功能

摘要

Objective:To study the synapse function of the neuron-like cells induced by rehmannia glutinosa polysaccharide.Methods:The bone marrow strornal stem cells (BMSCs) were cultured and purified by their characteristic of plastic adhesion,then induced by rehmannia glutinosa polysaccharide for 24 hours,and were cultured for 7 days.The membrane potential (MP),Ca2+ influx,synapse function were detected with laser-scanning confocal microscope.Results:BMSCs were induced for 24h,and cultured for 7 days,than neuron-like cells were observed to stretch out bumps and interact into complex network; Immunofluorescence cytochemistry demonstrated that the rate of nestin expression was 97.9％±1.3％,NSE expression 95.4％±1.9％ and the synaptophysin expression 94.2％ ±2.2％; the neuron-like cells were stimulated with high concentration KCl; the physiology chart showed that the curve rose shapely,Ca2+ influx increased and endocytosis and exocytosis happened.Conclusion:Rehmannia glutinosa polysaccharide can induce BMSCs to differentiate into the neuron-like cells with synaptic function.%目的:探讨地黄多糖诱导大鼠骨髓间充质干细胞(BMSCs)分化为神经元样细胞后是否具有神经突触功能.方法:贴壁筛选法分离纯化BMSCs,地黄多糖进行诱导,激光共聚焦显微镜检测细胞在高钾刺激下细胞膜电位的变化,细胞内钙流变化及细胞突触循环功能.结果:地黄多糖诱导24 h,连续培养7d后,光学显微镜下显示诱导后的细胞伸出突起交互成复杂网状；免疫荧光细胞化学显示诱导后的细胞神经元巢蛋白阳性表达率为97.9％±1.3％,神经元特异性烯醇化酶阳性率95.4％±1.9％,突触小泡蛋白阳性率为94.2％±2.2％；激光共聚焦显微镜显示诱导后细胞在高钾刺激下细胞膜电位迅速升高,细胞内钙离子流增加,细胞突触发生了胞吞胞吐现象.结论:地黄多糖可以诱导BMSCs分化为神经样细胞,此细胞具有神经细胞的神经生理功能.