代谢型谷氨酸受体7对人胚胎神经干细胞增殖的影响

目的 研究代谢型谷氨酸受体7(mGluR7)对人胚胎神经干细胞增殖的影响。方法 利用MTT比色法分析mGluR7过表达载体、mGluR7siRNA在不同时间对体外的人胚胎NSCs细胞活力的影响, 神经球测量方法分析mGluR7对人神经球生长的影响, 流式细胞术分析mGluR7对人NSCs细胞周期及细胞凋亡的影响。结果 MTT结果表明在处理24 h, 48 h和72 h后, mGluR7显著增强人NSCs的细胞活力, 增殖细胞数量显著增多, 神经球直径显著增大, mGluR7siRNA抑制人神经球的增殖。过表达mGluR7后, 与对照组相比G1/G0期和细胞比率显著下降, S期细胞比率显著上升, mGluR7过表达促进了细胞G1到S期转化, 沉默mGluR7将人NSCs细胞阻滞在G1/G0期。应用流式细胞术分析过表达mGluR7对人NSCs细胞凋亡的影响, 转染48 h后, 早凋晚凋均显著下降, 沉默mGluR7能够显著诱导人NSCs的早凋和晚凋。结论 mGluR7可促进体外培养的人胚胎皮质NSCs增殖。

Effect of metabotropic glutamate receptor 7 on proliferation of human embryonic neural stem cells

Objective This study is to investigate the effect of metabotropic glutamate receptor 7(mGluR7) on the proliferation of human embryonic neural stem cells (NSCs).Methods The pCMV2-GV146-GFP-mGluR7 plasmid was transfected to over-express mGluR7.while mGluR7 siRNA was transfected to knockdown mGluR7.MTT assay was used to analyze cell proliferation.Flow cytometry was used to detect cell cycle and apoptosis.Protein and mRNA levels were analyzed by Western blot and RT-qPCR, respectively.Results The viability of human NSCs and the diameter of neurospheres after 24hours, 48hours, and 72hours of transfection significantly increased by mGluR7 overexpression whereas significantly decreased by mGluR7 knockdown, indicating a promotive effect of mGluR7 on NSC proliferation.After mGluR7 overexpression, G1/G0 phase cell ratio dropped significantly compared with control group, while the S phase cell ratio increased.mGluR7 silencing arrested human NSCs at G1/G0 phase.After 48hours of transfection, there was a decrease of apoptosis by mGluR7 overexpression, while mGluR7 silencing induced apoptosis of human NSCs.Conclusion mGluR7 can promote the proliferation of human embryonic cortical NSCs in vitro.