脑组织中FoxO_(3)在高压氧暴露后的蛋白表达

FoxO_(3) protein expression in brain tissue after hyperbaric oxygen exposure

目的探讨小鼠暴露于高压氧后其中枢神经系统的反应和病理变化、FoxO_(3)蛋白的作用及其机制及FoxO_(3)对急性氧中毒的影响。方法将体重24~28 g的24只小鼠随机分为FoxO_(3)蛋白含量测定组和FoxO_(3) mRNA含量测定组,每组12只。FoxO_(3)蛋白含量测定组的12只小鼠再随机分为四组,在6个绝对大气压条件下暴露30 min,0、8、24 h分别取大脑皮层;FoxO_(3) mRNA含量测定组同样分组;检测FoxO_(3)蛋白在大脑皮质的表达及定位FoxO_(3)的位置,探讨FoxO_(3)起作用的信号、传导途径和机制。结果在大脑皮层中,Western blot检测FoxO_(3)蛋白表达中,正常组蛋白灰度值比为1.000,暴露结束后为1.128,结束后8 h为1.736,结束后24 h为1.504,结果表明结束后8 h含量达到峰值;免疫组化检测大脑皮层FoxO_(3)蛋白胞浆、胞核转位情况中,正常组细胞核阳性细胞比例为6%,暴露结束后为17%,结束后8 h为52%,表明暴露结束后8 h入核现象更加明显。结论FoxO_(3)可以明显减轻高压氧引起的急性中毒,以防止氧化应激过程中脑组织的伤害。

Objective To investigate the central nervous system reaction and pathological changes of mice exposed to hyperbaric oxygen,the role and mechanism of FoxO_(3) protein,and the effect of FoxO_(3) on acute oxygen toxicity.Methods Twenty-four mice with body weight from 24 to 28 g were randomly divided into the FoxO_(3) protein content determination group and the FoxO_(3) mRNA content determination group,with 12 mice in each group.Twelve mice in the FoxO_(3) protein content determination group were randomly divided into 4 groups:in the cerebral cortex of mice exposed to 6 absolute atmospheres(ATA)for 30 minutes were collected at 0h,8 h and 24 h,respectively.The FoxO_(3) mRNA content determination group was also divided into groups in this way.The expression of FoxO_(3) protein in cerebral cortex and the location of FoxO_(3) were detected to explore the signal,conduction pathway and mechanism of FoxO_(3) action.Results In the cerebral cortex,with FoxO_(3) protein expression detected by the Western blot,the gray value ratios of protein were 1.000 in the normal group,1.128 at 0 h after exposure,1.736 at 8 h after exposure,and 1.504 at 24 h after exposure,indicating that the content reached the peak at 8 h after exposure.Immunohistochemistry was used to detect the cytoplasmic and nuclear translocation of FoxO_(3) protein in the cerebral cortex.The proportions of nuclear positive cells were 6%in the normal group,17%at 0 h after exposure,and 52%at 8 h after exposure,indicating that nucleation was more obvious at 8 h after exposure.Conclusion FoxO_(3) can significantly reduce the acute toxicity caused by hyperbaric oxygen and prevent the damage of brain tissue during oxidative stress.