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目的:观察力竭运动前后大鼠脑皮质运动区NR2A蛋白含量及其自身酪氨酸磷酸化水平变化,分析二者之间的相关关系,为探讨中枢兴奋信号在运动中的传递机理以及运动性疲劳的中枢机制提供实验依据。方法:SD大鼠进行一次性力竭跑台运动。采用抗NR2A抗体和抗磷酸酪氨酸单抗以免疫沉淀法和免疫印迹法检测皮质运动区NR2A蛋白含量及酪氨酸磷酸化水平。结果:力竭运动后即刻,大鼠脑皮质运动区NR2A蛋白含量与安静组相比无显著性变化,运动后1小时与安静组和运动后即刻比较显著升高(P<0.05,P<0.01),运动后3小时NR2A蛋白含量下降,运动后恢复24小时大鼠脑皮质中NR2A蛋白含量显著低于安静组和运动后即刻组(P<0.01,P<0.05)。NR2A酪氨酸磷酸化水平力竭运动后与对照组相比呈升高趋势但无显著性差异,NR2A酪氨酸磷酸化水平与NR2A蛋白含量变化无显著相关。结论:(1)大鼠在力竭运动后即刻及恢复期过程中,大脑皮质运动区NR2A蛋白含量变化表现为下降,上升,再下降,表明运动过程中NR2A蛋白含量变化具有较敏感的可调控性,力竭运动后即刻NR2A蛋白含量的下降可能是导致中枢抑制的一个因素。(2)NR2A酪氨酸磷酸化水平在力竭运动后呈升高趋势,可能有利于维持中枢的兴奋性,提示运动过程中NR蛋白含量的变化可能是多方面原因造成的,受体蛋白含量与受体活性之间可能存在较为复杂的关系。
OBJECTIVE: To observe the changes of NR2A protein and tyrosine phosphorylation of cerebral cortex before and after exhaustive exercise in rats, and to analyze the relationship between the two. The central mechanism of fatigue provides experimental evidence. Methods: SD rats were subjected to one-time exhaustive treadmill exercise. NR2A protein and tyrosine phosphorylation were detected by immunoprecipitation and immunoblotting using anti-NR2A antibody and anti-phosphotyrosine monoclonal antibody. Results: Immediately after exhaustive exercise, the level of NR2A protein in cerebral cortex of rats did not change significantly compared with that of the quiet group. The level of NR2A protein in the cortex of the rats’ cortex increased significantly at 1 hour after exercise compared with that in the quiet group and immediately after exercise (P <0.05, P <0.01) ). The content of NR2A protein decreased 3 hours after exercise. The content of NR2A protein in cerebral cortex of rats recovered 24 hours after exercise was significantly lower than that of quiet group and immediate exercise group (P <0.01, P <0.05). NR2A tyrosine phosphorylation increased after exhaustive exercise compared with the control group but no significant difference, NR2A tyrosine phosphorylation and NR2A protein content was not significantly correlated. Conclusion: (1) The NR2A protein content in the motor cortex decreased, then increased and then decreased in rats during exhaustive exercise and convalescent phase, indicating that the NR2A protein content in the process of exercise is more sensitive and regulatable Decreased NR2A protein levels immediately after exhaustive exercise may be a contributing factor to central suppression. (2) The level of tyrosine phosphorylation of NR2A increased after exhaustive exercise, which may be helpful to maintain the excitability of the central nervous system. It indicates that the change of NR protein content during exercise may be caused by many factors. The content of receptor protein There may be a more complex relationship with receptor activity.