人参皂苷Rg3对体外培养小鼠神经干细胞增殖的影响
Effect of Ginsenoside Rg3 on Mouse Neural Stem Cells Proliferation in Vitro
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摘要: 目的 研究人参皂苷Rg3对体外培养小鼠神经干细胞 (NSCs) 增殖的影响.方法 分离培养胎鼠NSCs, 使用神经球悬浮培养法传代, 特异性抗体Nestin和Sox2鉴定NSCs.采用Brd U标记法和CCK-8法检测不同浓度Rg3 (20 n M、50 n M、250 n M和500 n M) 对NSCs增殖的影响.结果 传代2次后的神经球几乎全为Nestin/Sox2免疫荧光双阳性细胞, 表明其为纯度较高的NSCs.Brd U标记法表明与对照组、20 n M组、250 n M组和500 n M组相比, 50 n MRg3组Brd U阳性细胞相对比率最高, 差异有统计学意义 (P<0.01) , 但其他各组之间差异无统计学意义 (P>0.05) .CCK-8法检测, 结果显示50 n M Rg3在450 nm处吸光值最高, 且与对照组 (P<0.01) 、20 n M组 (P<0.05) 和500 n M组相比, 差异有统计学意义 (P<0.01) , 但其他各组之间无统计学差异 (P>0.05) .结论 人参皂苷Rg3能促进体外培养小鼠NSCs的增殖, 且其最佳浓度是50 n M.Abstract: Objective To investigate the effect of ginsenoside Rg3 on mouse neural stem cells (NSCs) proliferation in vitro.Me thods Fetal cortices of embryonic 14 d (E14) C57 BL/6 mouse were isolated for culturing primary NSCs.After being passaged twice, NSCs specific antibodies, Nestin and Sox2 were identified by immunocyto chemistry staining. CCK-8 assay and Brd U labelling assay were used to measure NSCs proliferation in response to different ginsenoside Rg3 concentrations (20 n M, 50 n M, 250 n M and 500 n M) . Re s ults Primary and passaged NSCs were successfully cultured and almost all the cells were both Nestin and Sox2 positive NSCs, which indicated high purity of NSCs. Brd U labelling assay showed the group of 50 n M Rg3 had the highest relative ratio of Brd U positive percentage, and the difference with other groups was statistically significant (P<0.01) .CCK-8 assay also showed 50 n M Rg3 group had the highest OD value, which was significantly higher than the control group (P<0.01) , 20 n M group (P<0.05) and 500 n M group (P<0.01) , while other groups had no significant difference between each other (P >0.05) . Conclus ion 50 n M ginsenoside Rg3 could significantly promote mouse NSCs proliferation in vitro.
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Key words:
- Neural stem cells /
- Ginsenoside Rg3 /
- Proliferation /
- BrdU /
- CCK-8
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