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作 者:柴丽娟[1] 钟佩茹[1] 周志焕[1] 王秀云[1] 石田寅夫 张艳军[1]
机构地区:[1]天津中医药大学,天津市中药药理学重点实验室,天津300193 [2]铃鹿医疗科学大学东洋医学研究所,日本铃鹿市510-0293
出 处:《中国药理学通报》2010年第5期670-673,共4页Chinese Pharmacological Bulletin
基 金:日本国文部科学省高技术研究中心整备事业项目(No16文科高第978号)
摘 要:目的探讨黄芪甲苷(astragaloside,AS)对神经干细胞增殖的影响。方法体外培养大鼠胎鼠神经干细胞,Nestin免疫化学染色鉴定NSCs,BrdU标记鉴定NSCs增殖。BrdU标记免疫细胞化学染色检测神经干细胞增殖能力。实时定量PCR技术探讨黄芪甲苷促进NSCs增殖的作用机制。结果Nestin鉴定为阳性,Brdu标记亦呈阳性;BrdU标记结果表明,黄芪甲苷高、中、低剂量组NSCs的增殖率明显提高(P<0.05,P<0.01)。实时定量PCR结果表明在诱导NSCs增殖过程中,黄芪甲苷高剂量组Hes5基因表达增加(P<0.05)。结论黄芪甲苷对NSCs增殖具有明显的诱导作用,其可能通过上调与细胞增殖相关基因Hes1、Hes5、cyclinD1表达而起作用,但也可能调节与其它增殖通路有关。Aim To explore the effects of astragaloside(AS)on the proliferation of neural stem cells(NSCs).Methods The NSCs of embryonic day(E)16 SD rat were cultured in vitro.The NSCs were identified by the immunocytochemical(ICC)staining of Nestin.The ICC staining of BrdU was adopted to characterize the proliferation of NSCs.The number of neurospheres and the ICC staining of BrdU were performed to identify their proliferation properties.Real time RT-PCR technique was used to investigate the proliferation mechanisms of the AS on the NSCs.Results Characteristic protein(Nestin)of NSCs labeling,BrdU labeling were positive showed by immunocytochemical staining.The ICC of BrdU labeling results showed that different dosages of AS could promote the proliferation of NSCs in vitro.The proliferation rate of NSCs was increased extremely significant(P〈0.05,P〈0.01).Real time RT-PCR analysis of Hes1,Hes5 and cyclinD1 demonstrated that Hes5 expression of high dosage group of AS(6 μmol·L^-1)increased significantly(P〈0.05).Conclusions Astragaloside significantly promotes the proliferation of NSCs in vitro.The real-time quantitative data suggest that Hes1,Hes5 and cyclinD1 might be involved in the proliferation of NSCs induced by astragaloside.
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