EDS对新生SD大鼠海马神经干细胞增殖分化的作用

Effects of ecdysterone on proliferation and differentiation of rat neural stem cells in vitro

目的观察在不同剂量蜕皮甾酮(ecdysterone,EDS)作用下,体外培养的神经干细胞(neuralstemcells,NSCs)增殖、分化情况。方法原代培养新生SD大鼠海马NSCs,经EDS处理后,传代细胞克隆形成率、四甲基偶氮唑盐比色法观察神经干细胞增殖能力的变化,免疫荧光细胞染色技术、流式细胞术检测神经干细胞分化情况。结果与对照组相比,中低剂量组(50、100、200mg/L),对大鼠海马NSCs增殖能力无显著影响,而高剂量的EDS(400mg/L及800mg/L)干预组则明显抑制了NSCs的增殖。在分化条件下,EDS200mg/L干预组显著提高NSCs向神经元分化的比例,其余各组无统计学差异。结论EDS能够调节体外培养的大鼠海马NSCs的增殖和分化水平,在合适的剂量下明显提高了神经元的分化比例。

Objective To explore the effects of ecdysterone （EDS） on proliferation and differentiation of rat hippocampal neural stem cells （NSCs） in vitro. Methods NSCs of SD rat hippocampus were isolated and cultured in serum-free medium containing bFGF and EGF, then different doses of EDS were used to treat the NSCs. Clone counting and MTT assays were performed for the proliferation of NSCs; Immunofluorescence and flow cytometry were carried out for the cell differentiation. Results High doses of EDS （400 and 800 mg/L） treatment markedly inhibited the proliferation of rat hippocampal NSCs, while the low doses of EDS （50, 100 and 200 ng/L） exhibited no effects on the NSCs proliferation. Treatment of NSCs with 200 mg/L EDS markedly increased the neurogenesis of NSCs in differential medium, while NSCs of other doses did not. Conclusion EDS regulates the proliferation and differentiation of NSCs in vitro, whereas only 200 mg/L EDS can increase the neurogenesis of rat hippocampal NSCs.