原代培养神经元细胞缺氧损伤后脑红蛋白表达研究

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—TimePCR、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、NgbmRNA及蛋白表达情况。结果原代培养皮层神经元缺氧4h后细胞生存率明显下降，NgbmRNA及蛋白表达升高，8h达高峰，此后逐渐稳步下降，至缺氧24h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。

Research of the Expression of Ngb Following the Hypoxia Insults in Primary Culture Cortical Neurons

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% 02 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 deter-mined 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 pro-tein were induced by hypoxia and began to increase in 4 hours to reach the peak in 8 hours , then de-clined slowly and steadily. The expression was still high than that of in control group in 24 hours. Con-clusion Ngb mRNA and protein can be induced by 1% O2 hypoxia continuing for at least four hours.