摘要
目的探讨姜黄素对神经干细胞内Wnt/β-catenin信号通路的影响。方法选择孕14.5 d的SD大鼠,取胚鼠大脑前端2/3部分组织,提取神经干细胞。培养并传代3次时,进行细胞免疫荧光学鉴定;同时换用只添加2%B27的Neurobasal培养基连续培养细胞,并设置空白对照组、IWR-1(Wnt信号通路特异性阻断剂10μmol/L)处理组、姜黄素500 nmol/L组、IWR-1+姜黄素500 nmol/L组(先添加IWR-1,20min后再加入姜黄素)、DMSO组。细胞培养3 d时,采用Western blot方法检测各组wnt3a、β-catenin蛋白表达水平变化,采用RT-PCR方法检测各组wnt3a、β-catenin的mRNA表达量变化。结果 Western blot结果显示:对培养第3代的细胞干预3 d后,姜黄素组wnt3a、β-catenin表达量较其他各组明显升高(P<0.05),IWR-1处理后其表达水平明显降低(P<0.05);姜黄素联合IWR-1处理细胞后wnt3a、β-catenin表达量明显减少(P<0.05);DMSO组表达量接近空白对照组。RT-PCR结果显示:与空白对照组比较,姜黄素组wnt3a及β-catenin基因表达水平明显上调(P<0.05),而IWR-1组wnt3a及β-catenin基因表达水平明显下降(P<0.05),总体变化趋势与Western blot结果一致。结论初步证明姜黄素可激活Wnt信号通路,由此推测姜黄素与经典的Wnt/β-catenin信号通路密切相关。
Objective To investigate the effect of curcumin on expression of Wnt/β-catenin signaling pathway in neural stem cells in vitro. Methods Neural stem cells were isolated from anterior two-thirds brains of fetal Sprague-Dawley （SD） rats （ 14.5 d after pregnancy）. The neural stem cells were passaged for 3 times by half media replacement and identified by cellular immunofluorescence assay. After 3 passages, cells were seeded in the media without bFGF or EGF. Then, cells were treated in different groups： control group, IWR-1 （a specific inhibitor to Wnt signaling pathway, 10 μmol/L） group, 500 nmol/L curcumin group, IWR-1 ＋ curcumin 500 nmol/L group and DMSO group. We measured Wnt3a and β-catenin expression levels by Western blotting and RT-PCR separately. Results Western blotting results showed that Wnt3a and β-catenin expression levels had significant increases in the curcumin group （P 〈 0. 05 ） , but were decreased in the IWR-1 group and IWR-1 ＋ cureumin group （P 〈 0. 05 ）. RT-PCR results showed that the expression levels of Wnt3a and β-catenin were increased in the curcumin group but decreased in the IWR-1 group （ P 〈 0. 05 ） , showing similar change pattern to the Western blot results. Conclusion Curcumin activates the classic Wnt/β-catenin signaling pathway in neural stem cells .
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2014年第8期764-768,共5页
Journal of Third Military Medical University
基金
国家自然科学基金(81070979
81000531)~~
作者简介
林江凯，电话：（023）68765251，E-mail：jklin@tmmu．edu．cn
