从线粒体动力学研究益气健脾活血法干预T2DM骨骼肌细胞凋亡的作用机制

Skeletal muscle is the major site of IR, which contains abundant mitochondrias, mitochondrial dynamics is involved in mitochondrial maintenance, biological productivity and cell death. The imbalance of mitochondrial dynamics is closely related to the occurrence of IR and T2DM. T2DM skeletal muscle ingury is related to function of spleen. Shenqi Compound Recipe from TCM theory mentioned above has the function of tonifying Qi, invigorating spleen and activating blood. It is clinically used to improve the patient muscle weakness, atrophy and pain. The preliminary study found that it had the function of improving glucose and lipid metabolism, reducing IR, anti-oxidative stress, improving inflammation, and improving the injury of skeletal muscle cells by regulating the cell apoptosis, but the mechanism is unclear. This topic will start from mitochondrial dynamics to observe after Shenqi compound intervention skeletal muscle mitochondria morphology change of rats with T2DM, measure rate of cell apoptosis, detect expression of mitochondrial dynamics, bcl-2 apoptosis protein family, Ras-Pi3k-Akt signal pathway related proteins and other apoptosis key factors in vitro and in vivo, and analysis the relevance of Mfn2 and mentioned related proteins, discuss exact mechanism of tonifying Qi, invigorating spleen and activating blood regulate the imbalance of mitochondrial dynamics to improve cell apoptosis of skeletal muscle of T2DM, to provide scientific basis of preventing and treating cell apoptosis of skeletal muscle of T2DM by tonifying Qi, invigorating spleen and activating blood.

骨骼肌是IR的主要部位，其内含有丰富的线粒体，线粒体动力学参与线粒体维护、生物产能及细胞死亡。线粒体动力学失衡与IR和T2DM的发生关系密切。在中医从脾论治糖尿病骨骼肌病变指导下研制的参芪复方具有益气健脾活血作用。临床中可明显改善患者肌无力、肌肉萎缩、肌肉酸痛。前期研究发现，该方可改善糖脂代谢、减轻IR、抗氧化应激、改善炎症状态，并通过调节细胞凋亡途径改善骨骼肌细胞损伤，但具体机制不明。本课题拟从线粒体动力学出发，通过体内外实验，观察参芪复方干预后T2DM大鼠骨骼肌细胞线粒体形态改变，测定细胞凋亡率，检测线粒体动力学、Bcl-2凋亡蛋白家族、Ras-Pi3k-Akt信号通路相关蛋白及其他凋亡关键因子的表达，分析Mfn2与上诉相关蛋白的关联性，探讨益气健脾活血法调节线粒体动力学失衡改善T2DM骨骼肌细胞凋亡的确切机制，为益气健脾活血法防治T2DM骨骼肌细胞凋亡提供科学依据。

骨骼肌是IR的主要部位，其内富含线粒体，线粒体动力学参与线粒体维护、生物产能及细胞死亡。线粒体动力学失衡与IR和T2DM的发生关系密切。在中医从脾论治糖尿病骨骼肌病变指导下组方的参芪复方具有益气健脾活血功效，可明显改善患者肌无力、肌肉萎缩、肌肉酸痛，但作用机制不明。本项目研究通过体内外实验，观察了参芪复方干预后T2DM大鼠骨骼肌细胞线粒体形态改变，测定细胞凋亡率，检测了线粒体动力学、Bcl-2凋亡蛋白家族、Ras-Pi3k-Akt信号通路相关蛋白及其它他凋亡关键因子的表达。本项目研究结果显示，益气健脾活血法可明显改善T2DM大鼠糖脂代谢，从基因和蛋白层面调节线粒体动力学失衡，减少骨骼肌细胞凋亡，发挥防治糖尿病骨骼肌病变的作用。本项目研究对揭示参芪复方防治T2DM骨骼肌病变的作用机制，对阐明益气健脾活血法科学内涵具有重要意义。