低氧预适应小鼠脑匀浆液提取液对大鼠鼠胚海马神经元缺氧复氧后神经细胞的影响

目的 探讨低氧预适应小鼠脑匀浆液提取液对大鼠鼠胚海马神经元缺氧复氧后神经细胞活性和凋亡的影响. 方法在96孔培养板中将大鼠鼠胚海马神经细胞原代培养至8 d,将培养细胞按照处理的不同分为以下5组:(1)正常对照组(仅加入PBS)、(2)H<,4>R<,48>组(缺氧4 h/复氧48h后加PBS)、(3)H0组(缺氧4 h/复氧48 h前加正常小鼠脑匀浆提取液)、(4)H1组(缺氧4h/复氧48h前加急性低氧对照小鼠脑匀浆提取液)、(5)H4组(缺氧4 h/复氧48 h前加低氧预适应小鼠脑匀浆提取液),分别用酶标仪和流式细胞仪测定神经细胞活性和凋亡情况.结果 正常对照组细胞活性明显高于H<,4>R<,48>组,H0、H1和H4组分别与H<,4>R<,48>组相比,细胞活性明显增加,且H4组细胞活性又明显高于H0、H1组;正常对照组仅有极少量的凋亡细胞,而H<,4>R<,48>组凋亡细胞显著增多,H0、H1和H4组分别与H<,4>R<,48>组相比凋亡细胞明显减少,且H4组又分别明显少于H0、H1组.结论低氧预适应小鼠脑匀浆液提取液可能通过增加大鼠鼠胚海马神经元缺氧复氧后神经细胞活性和减少神经细胞凋亡起到抗缺氧性损伤作用.

Effect of brain homogenate from hypoxia-preconditioned mice on rat embryonic hippocampal neurons with hypoxia/reoxygenation injury

Objective To observe the effect of brain homogenate (BH) extracted from hypoxia-preconditioned mice on the viability and apoptosis of rat embryonic hippocampal neurons with hypoxia/reoxygenation-induced injury. Methods Rat embryonic hippocampal neurons primarily cultured for 8 days in 96 well tissue culture plate were divided into 5 groups, namely the normal control group (treated with PBS), H<,4>R<,48> group (with hypoxia for 4 h followed by reoxygenation for 48 h and PBS treatment), H0 group (with BH from normal mice prior to hypoxia/reoxygenation), H1 group (with BH from acute hypoxia-preconditioned mice and hypoxia/reoxygenation), and H4 group (with BH from hypoxia-preconditioned mice and hypoxia/reoxygenation). The viability and apoptosis of the cells in the 5 groups were observed by MTT assay and flow cytomertry, respectively. Results The cell viability was significantly higher in the normal control group than in H<,4>R<,48> group. In H0, H1, and H4 groups, the cell viability increased significantly as compared with that in H<,4>R<,48> group, and the cells in H4 group showed the highest viability. Apoptotie cells were scarcely observed in the normal control group, but were numerous in H<,4>R<,48> group. Compared with H<,4>R<,48> group, H0, H1, and H4 groups showed obviously reduced apoptotic cells, and the reduction was the most conspicuous in H4 group. Conclusion BH extracted from hypoxia-preconditioned mice may offer protection against hypoxic injury of rat embryonic hippocampal neurons challenged with hypoxia-reoxygenation by promoting the cell viability and decreasing cell apoptosis.