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目的:研究芝麻素对NMDA所致原代培养小鼠皮层神经元损伤的保护作用及其机制。方法:体外培养原代皮层神经元;免疫荧光染色鉴定细胞纯度;MTT法测定细胞存活率;Hoechst/PI双染色观察细胞凋亡的形态学变化;激光共聚焦显微镜技术观察钙成像,检测细胞内钙离子浓度变化;Western blot检测各组细胞中Bcl-2、Bax、NR2A、NR2B蛋白的表达。结果:NMDA(200μM)60分钟能使神经元细胞存活率显著下降,细胞凋亡百分比明显增加(P<0.01),芝麻素(0.1μM)能提高细胞存活率,减少细胞凋亡(P<0.01)。与NMDA组相比,芝麻素能抑制钙超载;降低Bax和NR2B蛋白表达;增加和Bcl-2蛋白表达(P<0.01)。结论:芝麻素具有神经保护作用,这种作用可能与抑制钙超载、下调NMDA受体亚型NR2B的表达以及调节Bcl-2家族蛋白有关。
Objective: To study the protective effect of sesamin on cortical neurons injury induced by NMDA in mice and its mechanism. Methods: The primary cortical neurons were cultured in vitro. The purity of cells was identified by immunofluorescence staining. The cell viability was measured by MTT assay. The morphological changes of apoptosis were observed by Hoechst / PI double staining. Calcium imaging was performed by confocal laser scanning microscopy. The changes of Ca2 + concentration were detected by Western blot. The expressions of Bcl-2, Bax, NR2A and NR2B in each group were detected by Western blot. Results: NMDA (200μM) for 60 minutes could significantly decrease the survival rate of neurons and increase the percentage of apoptotic cells (P <0.01). Sesamin (0.1μM) increased the cell viability and decreased the apoptosis rate (P <0.01) ). Compared with NMDA group, sesamin could inhibit calcium overload, decrease Bax and NR2B protein expression, and increase Bcl-2 protein expression (P <0.01). Conclusion: Sesamin has neuroprotective effect, which may be related to the inhibition of calcium overload, down-regulation of NMDA receptor subtype NR2B expression and regulation of Bcl-2 family proteins.