HD02对新生大鼠海马神经干细胞增殖分化和细胞周期的影响

Effect of HD02 on proliferation/differentiation and cell cycle of hippocampus neural stem cells in newborn rats

目的:观察HD02对新生大鼠体外培养神经干细胞(neuralstemcell,NSC)增殖、分化和凋亡的影响。方法:分离、培养新生大鼠海马NSC,采用免疫荧光双标技术检测NSC增殖分化,流式细胞仪检测细胞周期和凋亡变化。结果:HD02的中低剂量组BrdU阳性细胞数、BrdU+Nestin,BrdU+NeuroD,BrdU+NF200,BrdU+GFAP,BrdU+Oli阳性细胞数和S期细胞比率明显高于空白对照组和EGB761组(P<0.01),G0/G1期比率(51.12±3.66)%明显低于正常对照组(68.23±3.30)%;细胞凋亡率(5.18±0.80)%与EGB761(4.98±0.40)%相比差异无显著性意义(P>0.05),但却明显低于正常对照组(6.43±0.55)%(P<0.01)。结论:HD02能显著促进NSC增殖分化。其机制可能与缩短G0/G1期、减少凋亡、促进S期比率有关。

AIM:To explore the effect of HD02 on proliferation,differentiation and apoptosis of neural stem cells(NSC) culture in vitro in newborn rats.METHODS:Neural stem cells of hippocampus in newborn rats were isolated and cultured.Immunofluorescence double-labeled technique was used to detect the proliferation and,differentiation of NSC,and flow cytometer to detect cell cycle and apoptosis.RESULTS:Compared with the normal control group and EGB761,the numbers of BrdU positive cells,BrdU+Nestin,BrdU+NeuroD,BrdU+NF200,BrdU+GFAP,BrdU+Oli positive cells and percentage of diploid synthesis phase(S phase) in the low and middle dosage groups of HD02 were significantly higher than those in the normal control group and EGB761 group(P< 0.01),the percentage of G0/G1(51.12±3.66)%was significantly lower than that in the control group(68.23±3.30)%,the cell apoptosis rate(5.18±0.80)%was not significantly different from that in the EGB761 group(4.98±0.40)%(P >0.05),but significantly lower than that in the control group(6.43±0.55)%(P< 0.01).CONCLUSION:HD02 can effectively promote the NSC proliferation and differentiation.The mechanism of HD02 may be related to the shortening of the period of G0/G1,reduction of cell apoptosis and promotion of S phase.