摘要
目的观察毛蕊花糖苷对原代成年小鼠神经干细胞增殖的影响,并探讨其可能的作用机制。方法从成年C57BL/6小鼠脑室下区分离、培养原代神经干细胞,并通过神经干细胞标志蛋白Nestin免疫荧光染色,对分离得到的神经干细胞进行鉴定。不同浓度毛蕊花糖苷(5、10、20、40μmol·L^(-1))在无有丝分裂原(EGF/bF GF)的条件下处理细胞24 h。采用CCK-8法检测细胞活力,免疫组化法计数BrdU阳性细胞率,检测细胞增殖能力,Western blot方法检测给药后神经干细胞Akt的磷酸化水平。结果毛蕊花糖苷在无有丝分裂原存在的条件下,能明显促进神经干细胞的增殖,并明显提高p-Akt的表达。而在加入PI3K/AKT信号通路阻断剂LY294002后,这一作用被明显抑制。结论毛蕊花糖苷对体外培养的神经干细胞具有明显促增殖作用,该作用机制可能与化合物激活Akt通路有关。
Aim To clarify the effect of acteoside on proliferation of neural stem cells( NSCs) from adult mice,as well as the involved signaling pathway. Methods NSCs were isolated from the subventricular zone( SVZ) of adult C57BL/6 mice,then identified by immunofluorescence staining with Nestin,the marker of NSCs. NSCs were exposed to acteoside( 5,10,20,40μmol·L^(-1)) in absence of mitogen( EGF/b FGF) for 24 h. We employed CCK8 assay to detect NSCs viability and Brd U staining to identify NSCs proliferation. We performed Western blot to quantify the expression level of p-Akt induced by acteoside on NSCs. Results Without mitogen,acteoside increased NSCs proliferation by activating p-Akt,which can be blocked by LY294002,the inhibitor of PI3 K / AKT signaling pathway. Conclusion Acteoside promotes the proliferation of NSCs from adult mice by activating PI3 K/AKT pathway.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第6期836-840,共5页
Chinese Pharmacological Bulletin
基金
天津市应用基础与前沿技术研究计划一般项目(No14JCYBJC42900)
关键词
毛蕊花糖苷
成年小鼠
神经干细胞
脑室下区
增殖
AKT通路
acteoside
adult mice
neural stem cells(NSCs)
the subventricular zone(SVZ)
proliferation
Akt pathway
