人参皂苷Rb1对体外培养胎鼠神经干细胞增殖及分化的影响

Intrathecal injection of carbenoxolone decreased the expression of CX43 and its effects on mirror-image pain in rats

目的:探讨人参皂苷Rb1对体外培养胎鼠神经干细胞(NSCs)增殖和分化的影响及其可能的作用机制。方法:分离培养胎鼠皮层NSCs,CCK-8法及BrdU掺入法测定不同浓度人参皂苷Rb1对NSCs增殖的影响,确定促进NSCs增殖的最佳人参皂苷Rb1浓度;细胞免疫荧光法检测各组β-Ⅲtubulin、GFAP及DAPI的表达,计算GFAP/DAPI、β-Ⅲtubulin/DAPI百分比;Western Blot、CCK-8法测定PI3K/Akt抑制剂LY294002干预下Akt、磷酸化Akt(p-Akt)蛋白表达水平及其OD值。结果:人参皂苷Rb1能够促进体外培养的NSCs增殖,最佳浓度为1μmol/L;不同浓度人参皂苷Rb1组GFAP/DAPI及β-Ⅲtubulin/DAPI百分比与对照组相比无显著性差异(P>0.05);但人参皂苷Rb1组(1μmol/L)的OD值及p-Akt蛋白水平均较对照组高(P<0.01),且这种作用可以被LY294002逆转。结论:人参皂苷Rb1在一定浓度范围内可以促进体外培养神经干细胞的增殖,PI3K/Akt信号通路在人参皂苷Rb1促进神经干细胞增殖作用中发挥重要作用。

Objective To explore the effects of ginsenoside Rbl on the proliferation and differentiation of rat embryoneural stem cells（NSCs） in vitro and the possible role mechanisms. Methods： NSCs were isolated and cultured from thecerebral cortex of embryonic rats. The proliferative ability of ginsenoside Rbl on NSCs was tested by CCK-8 assay andBrdU labeling, and then confirmed the optimal concentration, Immunofluorescenee cytochemistry was used to detect thepercentage of β-Ⅲ tubulin/DAPI and GFAP/DAPI. Under the pretreated of PI3K/Akt inhibitor LY294002, the level oftotal Akt/phosphorylated Akt （p-Akt） and OD value were evaluated by Western Blot, CCK-8, respectively. Results：The proliferation of NSCs cultured in vitro was observed to promote by Ginsenosdie Rbl, the optimal concentration of gin-senosdie Rbl was 1 μmol/L. There were no difference between the ginsenosdie Rbl and the control on the percentage ofGFAP/DAPI and β-Ⅲ tubulin/DAPI （ P 〉 0. 05 ）. But ginsenosdie Rbl could enhance the OD value and the expression ofp-Akt significantly （P 〈0.01 ）, and these effect were totally abolished by L.Y294002. Conclusion： Ginsenosdie Rblcould promote the proliferation of NSCs cultured in vitro in a centain concentration range, and PI3K/Akt pathway plays animportant role in this effect.