运动疲劳对小鼠海马GDNF、GFRα-1mRNA和蛋白表达的影响及益气养血补肾方的调节作用
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摘要
目的:探讨运动疲劳对小鼠海马组织GDNF、GFRα-1表达的影响及益气养血补肾方对运动疲劳的作用效果及机理。方法:将100只小鼠随机分为4组:对照组、模型组、西药组和中药组,除对照组外,其余三组以连续4周递增强度的无负重游泳的方式建立运动疲劳模型,并分别给予生理盐水、支链氨基酸水溶液、益气养血补肾方水煎液干预。分别用分光光度法检测血清及海马组织MDA含量,SOD、GSH-PX和iNOS活性,高效液相色谱法检测海马组织5-羟色胺(5-HT)和多巴胺(DA)含量,实时荧光定量PCR法检测海马组织胶质源性神经营养因子(GDNF)和受体GFRα-1 mRNA表达,免疫组化法检测GDNF和GFRα-1蛋白表达。结果:模型组小鼠海马GDNF、GFRα-1 mRNA和蛋白表达水平较对照组显著升高。中药组小鼠海马组织MDA、5-HT含量较模型组显著降低,DA含量、SOD活性及GDNF、GFRα-1mRNA和蛋白表达水平显著升高。结论:运动疲劳模型小鼠海马GDNF、GFRα-1mRNA和蛋白表达水平上调,提示GDNF和GFRα-1参与了运动疲劳产生的神经生物学调控过程。益气养血补肾方可以通过降低小鼠海马组织MDA、5-HT含量,提高DA含量、SOD活性,促进GDNF、GFRα-1的mRNA和蛋白表达水平而发挥神经保护作用,延缓或消除运动疲劳的发生。
Aim:To investigate GDNF and it's receptor GFRa-1 expression changes in mice hippocampus after exercise-induced fatigue and the effects and central mechanism of Tonifying Qi Nourishing the blood and Reinforcing Kidney Recipe on exercise-induced fatigue.
     Methods:100 healthy mice were randomly divided into control group, model group, the group of western medicine and the group of Chinese medicine, and established the model of exercise-induced fatigue through swimming without load of increasing interision.The lever of GDNF、GFRa-1 gene and protein expression, the concentration of 5-HT、DA、MDA, the activity of SOD、GSH-PX、iNOS were measured by Real-time Fluorescent Quantitative PCR、immunohistochemical method、high performance liquid chromatography (HPLC) and spectrophotometr method.
     Results:The mRNA and protein expression levels of GDNF and GFRa-1 were significantly higher in model group.5-HT, MDA levels were significantly lower in the group of Chinese medicine, furthermore DA levels, SOD activities, GDNF、GFRa-1 mRNA and protein expression levels were significantly increased in the group of Chinese medicine.
     Conclusion:Exercise-induced fatigues have increased the lever of GDNF and GFRa-1 mRNA and protein expression, which suggests that both GDNF and it's receptor GFRa-1 may involved in the neurobiological modulation for exercise-induced fatigue. Tonifying Qi Nourishing blood and Reinforcing Kidney Recipe may play a neuroprotective role in delaying or eliminating the occurrence of exercise-induced fatigue by reducing 5-HT, MDA content, increasing DA content, SOD activity, and GDNF、GFRα-1 mRNA and protein expression levels.
引文
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