【摘要】 目的探讨皮层神经元细胞缺氧损伤后脑红蛋白的表达。方法构建C57BL/6胎鼠皮层神经元原代培养的细胞缺氧模型,联合MTT微量酶反应比色法、Real-Time PCR、免疫荧光染色及Western Blot等方法,观察缺氧后细胞形态学、细胞生存率、Ngb mRNA及蛋白表达情况。结果原代培养皮层神经元缺氧4 h后细胞生存率明显下降,NgbmRNA及蛋白表达升高,8 h达高峰,此后逐渐稳步下降,至缺氧24 h仍高于对照组。结论缺氧损伤可诱导原代培养皮层神经元NgbmRNA及蛋白的表达。更多还原

【Abstract】 Objective To investigate the expression of Ngb following the hypoxia insults in cortical neurons.Methods Primary mouse cortical neurons were isolated from 16-day embryonic cortex obtained from the pregnant C57BL/6 mouse.The cultured neurons were suffered from 1%O2 hypoxia for different time.The morphological changes were compared between control and hypoxia groups.Cell viability was measured by MTT assay,Ngb mRNA was measured by Real-time PCR,and the Ngb protein was determined by immunofluorescence staining and Western Blot analysis.Results The morphology was changed and cell survival was inbited by 1% O2 hypoxia in primary culture cortical neurons.NgbmRNA and protein were induced by hypoxia and began to increase in 4 hours to reach the peak in 8 hours,then declined slowly and steadily.The expression was still high than that of in control group in 24 hours.Conclusion Ngb mRNA and protein can be induced by 1% O2 hypoxia continuing for at least four hours.更多还原