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目的:观察姜黄素对神经干细胞增殖和凋亡的影响,并探讨其作用机制是否与PI3K/AKT信号通路有关。方法:分离新生24 h昆明小鼠的海马组织,提取原代神经干细胞并培养,并用免疫荧光检测鉴定。收集第3代神经干细胞并随机分为空白组、缺血模型组及不同浓度姜黄素干预组(0、0.1、0.5、2.5、12.5及62.5μmol/L),通过3-(4,5)-2-噻唑-(2,5)-二苯基溴化四氮唑蓝药敏试验(MTT)检测姜黄素干预后神经干细胞的存活情况,5-溴脱氧尿嘧啶核苷(Brdu)增殖试剂盒检测神经干细胞增殖情况。利用免疫蛋白印迹(Western blotting)法检测不同浓度姜黄素干预后神经干细胞pPI3K、AKT蛋白的表达情况。结果:观察到立体感强且折光好的神经球。MTT结果显示:姜黄素可增强神经干细胞增殖能力,并具有浓度和时间依赖性。Brdu检测结果显示:随着姜黄素浓度增加和干预时间延长,神经干细胞凋亡逐渐受抑制。姜黄素可上调PI3K、AKT蛋白的表达,并随着浓度逐渐上调,增强作用更加明显。结论:姜黄素可促进神经干细胞增殖,抑制其凋亡,这可能与激活PI3K/AKT信号通路有关。
Objective: To observe the effect of curcumin on the proliferation and apoptosis of neural stem cells and to explore whether its mechanism of action is related to the PI3K / AKT signaling pathway. Methods: Hippocampal tissues of neonatal Kunming mice were isolated for 24 h. Primary neural stem cells were harvested and cultured. The cells were identified by immunofluorescence. The third generation neural stem cells were collected and randomly divided into blank group, ischemia model group and curcumin intervention group (0, 0.1, 0.5, 2.5, 12.5 and 62.5μmol / L) The survival rate of neural stem cells after treatment with curcumin was tested by MTT assay. The proliferation of 5-bromodeoxyuridine (Brdu) proliferation kit Detection of neural stem cell proliferation. Western blotting was used to detect the expression of pPI3K and AKT protein after intervention with different concentrations of curcumin. Results: observed three-dimensional strong and refraction of good neurospheres. MTT results showed that curcumin can enhance the proliferation of neural stem cells in a concentration-dependent and time-dependent manner. Brdu test results showed that: with the increase of curcumin concentration and prolonged intervention, apoptosis of neural stem cells gradually inhibited. Curcumin can up-regulate the expression of PI3K, AKT protein, and gradually increase with the concentration, the enhancement effect is more obvious. Conclusion: Curcumin can promote the proliferation and inhibit the apoptosis of neural stem cells, which may be related to the activation of PI3K / AKT signaling pathway.