低氧对大鼠大脑皮层细胞表达脑源性神经生长因子的影响

目的:观察不同氧浓度环境下不同时间处理对大脑皮层细胞分泌脑源性神经生长因子(BDNF)及表达BDNF mRNA的影响。方法:原代培养SD大鼠大脑皮层细胞,5 d后将低氧组细胞转移至低氧工作站,在1%和4%氧浓度环境下分别处理3 h和6 h,其相应的常氧对照组细胞则继续放置于CO2孵箱中。ELISA法检测细胞培养液中BDNF蛋白含量;qPCR法检测细胞内BDNF mRNA表达水平。结果:与对照组相比,4%和1%氧浓度环境3 h组细胞BDNF mRNA及蛋白表达无明显变化(P>0.05),但均有增加趋势;而6 h组均引起BDNF mR-NA及蛋白表达明显增高(P<0.05)。结论:低氧可诱导大鼠大脑皮层细胞的BDNF分泌和BDNF mRNA表达增加,且皮层细胞的反应性与低氧浓度和低氧刺激时间有关。

Effect of hypoxia on brain derived neurotrophic factor mRNA and protein expressions in cerebral cortex cells of rats

Objective: To observe the effect of different oxygen concentration on the expressions of brain derived neurotrophic factor(BDNF) mRNA and protein in cerebral cortex cells under different treating time.Methods: The primary cerebral cortex cells of SD rats were designed into two hypoxia groups and two normoxia control groups after 5 days.Then,the cells of hypoxia groups were treated for 3 hours and 6 hours under the environment of 1% and 4% oxygen concentration respectively in hypoxia workstation,while the corresponding control groups remained in HHCP continually.The ELISA method was used to detect BDNF protein content in cultural supernatants,and the level of intracellular BDNF mRNA was assayed by qPCR.Results: Compared to the control groups,the level of BDNF mRNA and protein of 3 hours groups had no significant difference under the environment of 1% and 4% oxygen concentration(P>0.05),but had increase trend,while the level of 6 hours groups were up-regulated obviously(P<0.05).Conclusion: The expressions of BDNF mRNA and protein in cerebral cortex cells can be induced by hypoxia,and the response of cerebral cortex cells is related with the concentration and treatment time of hypoxia.