亚健康疲劳(FSH)已成为严重的公共卫生问题，然而目前对其知之甚少。课题组前期研究表明：能量代谢紊乱在亚健康疲劳发生发展过程中起到重要作用。进一步研究发现亚健康疲劳小鼠骨骼肌细胞中出现线粒体自噬的过度活化，而防治亚健康疲劳有确切疗效的中药复方维康颗粒能够抑制这种变化。因此我们推测细胞自噬过度可能是亚健康疲劳能量代谢紊乱的关键环节，而调节细胞自噬活性也可能是维康颗粒的重要药理机制。本项目拟首先应用亚健康疲劳小鼠模型，利用特异性激动剂、抑制剂，分别激活、抑制细胞自噬，以确定细胞自噬在亚健康疲劳发生中的作用；其次利用体外细胞模型，采用基因转染、RNA干扰等方法，阐明亚健康疲劳细胞自噬调控的关键信号传导通路；最后，应用血清药理学方法，明确维康颗粒对亚健康疲劳细胞自噬的调节效应。这不仅促进亚健康疲劳的防治和中医"治未病"的发展，还将深化理解细胞自噬的生物学功能，同时开辟中西医结合研究新的思路和途径。

The results of epidemiological investigation showed that fatigue of sub-health (FSH) has become a serious public health problem. However, the pathogenesis of FSH is not yet fully clarified. Our previous studies have suggested that disorder of energy metabolism may play an important role in the development of FSH. Furthermore, excessive autophagy of Mitochondrial was found in skeletal muscle cells of FSH mouse model. In addition, traditional Chinese medicine Weikang granules could inhibit the excessive autophagy. Therefore, we hypothesized that excessive autophagy may play an important role in the disorder of energy metabolism and Weikang granules could be effective in the prevention and treatment of FSH by regulating autophagy. The steps of this project are as following. Firstly, we activate or inhibit the process of autophagy in the mouse model of FSH to make sure that autophagy plays an important role in the development of FSH. Secondly, we apply the technology of gene transfection and siRNA in vitro to find out the critical signal transdction pathway of regulating autophagy. Finally, we use the method of serum pharmacology to clarify the effects of Weikang granules on regulating autophagy. This is of great significance not only for the prevention and treatment of FSH, but also deepen the understanding of the biological functions of autophagy.

中药复方维康颗粒防治亚健康疲劳疗效确切，但其机制尚未阐明。本项目拟探讨细胞自噬在亚健康疲劳发生发展中的作用及其mTOR信号通路的调控机制，明确维康颗粒对亚健康疲劳细胞自噬的调节效应，为其临床应用提供科学依据。应用亚健康疲劳小鼠和骨骼肌细胞模型，利用特异性激动剂、抑制剂或过表达、沉默mTOR，分别激活、抑制细胞自噬，采用负重力竭游泳时间测定小鼠疲劳程度，酶标仪测定氧化应激指标MDA、SOD、TAC水平，透射电镜观察组织细胞超微结构，CCK-8法检测细胞活力，分光光度仪检测线粒体COX活性，HPLC检测线粒体能量负荷值，MDC法检测细胞自噬水平，Western blot检测相关蛋白表达情况。结果表明，自噬抑制剂或过表达mTOR能够改善亚健康疲劳状态，减轻氧化应激损伤，提高亚健康疲劳骨骼肌细胞活力和COX活性以及能量负荷值，而降低细胞自噬水平；维康颗粒对亚健康疲劳小鼠有确切的防治作用，激活mTOR信号通路，提高亚健康疲劳骨骼肌细胞活力而降低细胞自噬水平，保护线粒体结构和功能。结论：线粒体自噬在亚健康疲劳发生发展过程中起到重要作用，mTOR信号通路参与亚健康疲劳线粒体自噬的调节过程，而维康颗粒可通过激活mTOR信号通路抑制线粒体自噬而改善亚健康疲劳状态。这促进了亚健康疲劳的防治和中医“治未病”研究。