Icariin promotes cell proliferation and regulates gene expression in human neural stem cells in vitro
Icariin promotes cell proliferation and regulates gene expression in human neural stem cells in vitro
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淫羊藿苷体外促进人神经干细胞增殖并调节基因表达
DOI：
10.3892/mmr.2016.5377
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发表时间：
2016-08-01
影响因子：
3.4
通讯作者：
Li, Lin
中科院分区：
文献类型：
--
作者：
Yang, Pan;Guan, Yun-Qian;Li, Lin
淫羊藿苷（Icariin， ICA）是从淫羊藿中提取的重要生物活性成分，具有神经保护作用。本研究旨在探讨ICA对体外培养的人神经干细胞（NSCs）细胞增殖和基因表达的调控作用。从自然流产后获得的16-20周人胎纹状体中分离单细胞。细胞在Dulbecco改良Eagle培养基/F12全培养基中培养，采用免疫染色和细胞分化实验进行鉴定。用ICA处理NSCs，使用cell Counting kit-8细胞增殖测定试剂盒评估细胞增殖。此外，还比较了ica处理细胞与对照细胞的神经球形成情况。采用基因芯片分析检测ICA对基因表达的影响。通过定量聚合酶链反应（qPCR）进一步分析NSC增殖重要基因表达的改变。结果表明，纹状体分离细胞培养7-10天后出现典型的神经球。这些细胞表达巢蛋白，一种重要的NSC标记物，在分化培养基存在下，它们表达β - iii -微管蛋白，一种特定的神经元标记物，和胶质纤维酸性蛋白，一种星形胶质细胞标记物。ICA能促进NSC的增殖和神经球的形成。微阵列数据和通路分析显示，ICA调控的基因参与多种信号通路，包括Wnt和碱性成纤维细胞生长因子（bFGF）通路，这些通路对NSC功能的调控具有重要意义。通过qPCR进一步验证了Wnt通路中关键角色卷曲类受体7和凌乱段极性蛋白3以及bFGF受体成纤维细胞生长因子受体1的上调和Wnt通路抑制剂糖原合成酶激酶3 β的下调。综上所述，ICA促进人NSCs增殖并调节基因表达，提示ICA可能通过调节NSC活性发挥其神经保护作用。
Icariin (ICA), which is an essential bioactive component extracted from the herb Epimedium, possesses neuroprotective properties. The aim of the present study was to investigate the regulatory roles of ICA in cell proliferation and gene expression in human neural stem cells (NSCs) in vitro. Single cells were isolated from the corpus striatum of 16-20-week human fetuses obtained following spontaneous abortion. The cells were cultured in Dulbecco's modified Eagle's medium/F12 complete medium and were characterized by immunostaining and cell differentiation assay. NSCs were treated with ICA, and cell proliferation was assessed using the Cell Counting kit-8 cell proliferation assay kit. In addition, neurosphere formation was comparatively studied between the ICA-treated and control cells. cDNA microarray analysis was performed to examine the effects of ICA on gene expression. Altered expression of genes important for regulating NSC proliferation was further analyzed by quantitative polymerase chain reaction (qPCR). The results demonstrated that typical neurospheres appeared after 7-10 days of culturing of individual cells isolated from the corpus striatum. These cells expressed nestin, an important NSC marker, and in the presence of differentiation medium they expressed beta-III-tubulin, a specific neuronal marker, and glial fibrillary acidic protein, an astrocyte marker. Treatment with ICA enhanced NSC proliferation and the formation of neurospheres. Microarray data and pathway analysis revealed that the genes regulated by ICA were involved in several signaling pathways, including the Wnt and basic fibroblast growth factor (bFGF) pathways, which are important for the regulation of NSC function. Upregulation of frizzled class receptor 7 and dishevelled segment polarity protein 3, which are key players in the Wnt pathway, and fibroblast growth factor receptor 1, which is the receptor for bFGF, and downregulation of glycogen synthase kinase-3 beta, which is a Wnt pathway inhibitor, was further validated by qPCR. In conclusion, ICA promoted proliferation and regulated gene expression in human NSCs, thus suggesting that ICA may exert its neuroprotective effects by regulating NSC activity.