黄芪注射液对人羊膜上皮细胞向神经细胞分化作用及存活的影响

目的:探讨黄芪注射液体外定向诱导人羊膜上皮细胞分化的作用和黄芪注射液对分化细胞活力影响。方法:将人羊膜上皮细胞分为三组,即黄芪诱导组(设立四个浓度亚组)、全反式维甲酸组和对照组。分别采用化学诱导剂和黄芪注射液诱导hAECs分化为神经细胞,在诱导前后,应用免疫细胞化学方法染色鉴定神经元特异性烯醇化酶、神经干细胞巢蛋白、胶质纤维酸性蛋白.逆转录-聚合酶链反应进一步鉴定细胞多能基因Oct4、神经干细胞标记物Nestin基因和神经元标记物NSE,四甲基偶氮唑盐比色法观察细胞活力。结果:倒置显微镜下见,RA诱导12 h后,黄芪诱导24 h后,由胞体伸出较长的轴突样和树突样突起,且有分支,48 h时100μl/ml黄芪诱导组,NSE阳性率低于RA诱导组(P<0.05),细胞活力较RA组高(P<0.05),两诱导组结果均高于对照组,而GFAP阳性率高于RA组(P<0.05),同时RT-PCR检测到诱导后多能基因Oct4表达减少,而Nestin、NSE表达增多。结论:黄芪和RA都可诱导人羊膜上皮细胞在体外分化为神经元样细胞,且黄芪于100μl/ml浓度时效果较好,黄芪诱导后对细胞毒性较小。

The effects of astragalus on the differentiation and survival of human amniotic epithelial cells to neurocytes

Objective: To explore the effects of astragalus on human amniotic epithelial cells differentiate to neuron-like cells in vitro and survival effect of hAECs.Methods: hAECs were randomly divided into astragalus group(4 subgroups),RA group and control group.Astragalus group and RA group were induced to differentiate into neurocytes by using chemical inducer RA and astragalus.The expression of NSE,Nestin and GFAP of induced cells in three groups were showed using immunocytochemical method.RT-PCRwas further used to detect the expression of multiple genes Oct4,Nestin,NSE.The cell viability was measured by methyl thiazolyl tetrazolium methods.Results: Under the convert microscope it was observed that hAECs started to change their shape,and there were several axon or dendrite-like processes out from the cell body induced with astragalus after 24 hours or RA after 12 hours,the positive expression of the NSE cells in 100 μl/ml astragalus-induced group was less than that in RA-induced group after 48h(P<0.05),But the positive expression of the GFAP cells in 100 μl/ml astragalus-induced group was higher than that in RA-induced group after 48 h(P<0.05),Results in the two groups were higher than those in control group,cell viability higher than those of RA group(P<0.05).RT-PCR showed the expression of Oct4 was less than control group,but Nestin and NSE were higher than control group.Conclusion: Astragalus(especial at 100 μl/ml-48h) and RA can induce hAECs into neuron-like cells,Astragalus-indoced group has a higher cell survival rate than RA-induced group.