甘草苷对小鼠胚胎脑神经干细胞增殖的影响  

Effect of liquiritin on the proliferation of neural stem cells from the brain of mouse embryos

作　　者：李社芳[1] 苗灵娟[2] 李宁[3] 梁鹤[3] 任德启[4] 郭健[4] 

机构地区：[1]河南中医药大学第一临床医学院内科学科,河南省郑州市450000 [2]河南省中医药研究院附属医院高血压科,河南省郑州市450004 [3]河南中医药大学基础医学院,河南省郑州市450046 [4]河南中医药大学第二临床医学院,河南省郑州市450008

出　　处：《中国组织工程研究》2017年第21期3332-3337,共6页Chinese Journal of Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(122300410529);课题名称:中药复方对脑胶质瘤干细胞分化;增殖的实验研究~~

摘　　要：背景:甘草苷对神经细胞有保护和营养作用,但甘草苷对小鼠胚胎脑神经干细胞增殖的影响尚未见报道。目的:研究不同质量浓度甘草苷对小鼠胚胎脑神经干细胞增殖的影响。方法:从孕14 d昆明白小鼠胚胎脑组织中分离出神经干细胞并培养,采用免疫细胞化学方法鉴定神经干细胞,利用qR T-PCR检测神经干细胞特异性基因表达,MTT法绘制神经干细胞生长曲线。取生长旺盛的神经干细胞,分别以0(对照),1,2,4,8 g/L的甘草苷干预,干预48 h后,MTT法检测细胞增殖。结果与结论:(1)培养第5天时,所有单个的神经干细胞全部聚集成球,随着时间的延长,细胞球体积越来越大,细胞球融合成更大的细胞团;(2)免疫细胞化学鉴定显示,100%的神经干细胞球呈巢蛋白阳性表达;(3)qR T-PCR检测显示,神经干细胞巢蛋白阳性表达较强,不表达神经元特异性烯醇化酶及胶质纤维酸性蛋白;(4)神经干细胞刚开始生长缓慢,处于生长缓慢期,第二三天起,细胞增殖迅速,进入指数生长期;第4天后,细胞增殖逐步减缓,细胞整体进入生长平台期;(5)2,4,8 g/L甘草苷组细胞增殖明显高于对照组(P<0.05),并且随着甘草苷质量浓度的升高,神经干细胞增殖速度逐渐升高;(6)结果表明,2-8 g/L甘草苷能促进小鼠胚胎脑神经干细胞的增殖。BACKGROUND：Liquiritin has the protective and nutritive effects on neural stem cells. However, the effect of liquiritin on neural stem cells from the brain of mouse embryos remains unclear. OBJECTIVE：To investigate the effects of different concentrations of liquiritin on the proliferation of neural stem cells from the brain of mouse embryos. METHODS：Neural stem cells were separated from the embryonic brain of Kunming white mice at the gestational age of 14 days. The identification of embryonic neural stem cells was performed by immunocytochemistry method. The expression of neural stem cells-special genes was determined by qRT-PCR. The cell growth curve was drawn and proliferation of embryonic neural stem cells treated with 0, 1, 2, 4 or 8 g/L liquiritin for 48 hours was measured by MTT assay. RESULTS AND CONCLUSION：（1） When cultured at day 5, al individual neural stem cells gathered together into neurospheres; with the extension of time, the neurospheres were enlarged, and gathered together into larger cell masses. （2） Results from immunocytochemistry showed that all the floating neurospheres was nestin-positive. Data from qRT-PCR revealed a higher expression of nestin mRNA, but there was no expression of neuron-specific enolase and glial fibrillary acidic protein in the neural stem cells. （4） The growth of neural stem cells was slow at the beginning. After 2-3 days, the cell proliferation quickly entered the exponential phase. After 4 days, the cell proliferation gradually slowed down, and the overall cell growth entered into the platform period. （5） The cell proliferation after treatment with 2, 4 or 8 g/L liquiritin was faster than that in the control group （0 g/L）. To conclude, 2-8 g/L liquiritin could increase the proliferation of neural stem cells from the brain of mouse embryos.

关 键 词：神经干细胞 中草药 细胞增殖 组织工程 干细胞 胚胎干细胞 胚胎脑神经干细胞 神经系统 甘草苷 神经保护 NESTIN 增殖 

分 类 号：R394.2[医药卫生—医学遗传学]