高温对小鼠神经元骨架蛋白及热休克蛋白70表达的影响
作者：耿晓英，万琪，杨荣军，吴松笛【关键词】 高温；神经元；骨架蛋白；热休克蛋白质70
【Abstract】 AIM: To observe the effects of hyperthermia (39℃) on the expressions of skeleton protein (βtubulin) and heat shock protein 70 (HSP70) of cortical neurons in vitro cultured and explore the neuroprotective effect of HSP70. METHODS: The cerebral cortical neurons of embryonic mice were cultured. The cultured neurons were divided as control group which was cultured at 37℃ and hyperthermia group which was put in high temperature (39℃) for 30 min. The immunofluorescence and laser scanning confocal imaging were used to detect HSP70 and βtubulin expressions at different time points (0, 2, 6, 24, 72 h) after hyperthermia. RESULTS: The fluorescence intensity of βtubulin in hyperthermia group was lower than that in control group, and it increased as recovery time went along. The fluorescence intensity of HSP70 in former group was higher than that in the latter, and it increased firstly and then decreased. The level of HSP70 reached the highest level at 2 h and remained a high level for 24 h. The expression of HSP70 was in nucleus after hyperthermia and then in cytoplasm as time went on. CONCLUSION: Hyperthermia may lead to the structural change of the cerebral cortical neuron probably through influencing skeleton protein structure. HSP70 may be involved in the protection of neurons in hyperthermia.
【Keywords】 hyperthermia; neurons; skeleton protein; Heat shock proteins 70
【摘要】 目的： 观察39℃环境对体外培养的小鼠皮层神经元骨架蛋白(βtubulin)和热休克蛋白70（HSP70）表达的影响，探讨HSP70的神经保护作用. 方法： 取小鼠胚胎大脑皮层进行神经元原代培养，将培养好的神经元分为37℃培养的对照组和39℃刺激30 min后0, 2, 6, 24和72 h的高温组. 分别进行荧光免疫染色，应用激光共聚焦扫描观察神经元βtubulin和HSP70的表达. 结果： 39℃刺激后，高温组神经元βtubulin荧光强度均低于37℃对照组，且随着恢复时间的'延长其荧光强度值逐渐升高. 高温组神经元HSP70荧光强度均高于37℃对照组，且随着恢复时间的延长其荧光强度值先升高后降低，于2 h时达高峰，24 h仍维持在较高的水平. 高温可导致HSP70荧光分布转移至细胞核，并随着恢复时间的延长逐渐转移至细胞浆. 结论： 高温可以导致神经元形态变化，骨架蛋白结构紊乱可能是神经元形态变化的重要原因之一，HSP70可能参与并起到保护作用.
【关键词】 高温；神经元；骨架蛋白；热休克蛋白质70
0引言
高温环境下，当人体内温度上升到40℃以上时，就会出现中枢神经系统功能障碍如：谵妄、惊厥或昏迷，甚至死亡. 但其复杂的详细机制目前仍不清楚. 机体在高温环境下受到热应激时，正常的蛋白质合成会受到抑制，但一组高度保守的热休克蛋白（heat shock proteins, HSPs）会迅速合成［1］. 热休克蛋白70（heat shock protein 70, HSP70）是热休克蛋白家族中的主要成员之一，是重要的分子伴侣. 在各种细胞器中，核仁和细胞膜对热刺激特别敏感，HSP70广泛存在于细胞骨架和核骨架内. 为了探讨高温环境下神经元HSP70和βtubulin表达的变化，我们采用体外培养的神经元，模拟高温环境，应用激光共聚焦扫描方法观察了高温刺激对神经元HSP70和βtubulin表达的影响，并初步探讨其可能的生物学意义.
1材料和方法
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