优化新生脉散方提高心衰大鼠运动耐量效应的非心源机制研究

The common manifestation of the patients with heart failure is the decreasing exercise tolerance, ventricular remodeling is the main reason, and skeletal muscle atrophy is also an important factor, but only by improving ventricular remodeling can not be equal exercise tolerance also received a corresponding improvement. Previous study indicates that with the addition of optimization new Shengmaisan Powder for promoting blood circulation prescription could further improve heart failure patients’ cardiac function,exercise tolerance and quality of life,but the lack of depth Mechanism .Ubiquitin-proteasome system participates in controling the degradation of protein within the cell and adjusting of apoptosis.Airogln-1/MAFbx and Murf-1 play important roles in myocardial hypertroph, apoptosis and skeletal muscle atrophy.They are the specifically expressed ubiquitin-ligating enzyme which stores in musculus cardiacus and osteomuscular. This study proposed the anterior descending coronary artery of rats by ligation method to establish rat model of heart failure, using cardiac ultrasound and swimming exhaustion test to evaluate rat cardiac function and exercise tolerance, using inTUNEL、 Western blot and RT-PCR to detect Airogln-1/MAFbx、Murf-1 protein expression and mRNA expression.From traditional Chinese medicine and the overall effect of the target, the study discusses the Non-Cardiac Mechanism of optimization new Shengmaisan Powder for promoting blood circulation prescription for improving function and exercise tolerance.

运动耐量下降是心衰患者最常见的临床表现，心室重构是其主要原因，而骨骼肌病变也是重要因素，仅通过改善心室重构并不能使运动耐量也获得相应的改善。前期研究显示优化新生脉散方可明显改善心衰患者的心功能、提高运动耐量，但缺乏深入机制探讨。泛素-蛋白酶体通路（UPS）可调控蛋白质的降解及激活JNK信号通路参与细胞凋亡。心肌及骨骼肌均含特异表达的泛素连接酶Airogln-1/MAFbx及Murf-1，可调控肌钙/球蛋白降解和细胞凋亡。本研究基于对UPS的表达调控，通过结扎大鼠冠脉前降支建心衰模型，采用心脏超声、力竭实验、力传感器等评价心衰大鼠心功能及运动耐量，TUNEL、荧光免疫、Western Blot、RT-PCR等技术检测细胞凋亡及UPS通路中Airogln-1/MAFbx、Murf-1等蛋白和mRNA的表达，从中医药多靶点整体效应角度，探讨优化新生脉散方提高运动耐量的非心源机制。

运动耐量下降是心衰患者最常见的临床表现，心室重构是其主要原因，而骨骼肌病变也是重要因素，仅通过改善心室重构并不能使运动耐量也获得相应的改善。前期研究显示优化新生脉散方可明显改善心衰患者的心功能、提高运动耐量，但缺乏深入机制探讨。泛素-蛋白酶体通路（UPS）可调控蛋白质的降解及参与细胞凋亡。心肌及骨骼肌均含特异表达的泛素连接酶Airogln-1/MAFbx及Murf-1，可调控肌钙/球蛋白降解和细胞凋亡。本研究基于对UPS的表达调控，通过结扎大鼠冠脉前降支建心衰模型，采用心脏超声、力竭实验、多道生理仪评价心衰大鼠心功能及运动耐量，采用苦味酸-天狼星红(SR)染色、TUNEL、荧光比色法、免疫组化等技术检测细胞凋亡及UPS通路中Airogln-1/MAFbx、Murf-1等蛋白表达，同时探索性的对心衰大鼠心肌组织进行泛素化修饰蛋白质组学研究，从中医药多靶点整体效应角度，探讨优化新生脉散方提高运动耐量的非心源机制。研究一：优化新生脉散方对心衰大鼠运动耐量影响的研究。结果显示：与模型组相比，优化新生脉散方可明显提高心衰大鼠LVEF、LVFS、±dP/dt max (mmHg/s) 、力竭游泳时间，差异具有统计学意义（P＜0.01），提示优化新生脉散方可提高心衰大鼠的心功能及运动耐量。研究二：优化新生脉散方提高心衰大鼠运动耐量的分子生物学机制的研究，结果显示：与模型组相比，优化新生脉散方可明显抑制心肌、腓肠肌和膈肌组织中Airogln-1/MAFbx、Murf-1蛋白表达，改善心衰大鼠心肌、腓肠肌和膈肌的细胞凋亡，抑制心肌纤维化，差异具有统计学意义（P＜0.01）。同时探索性的对心衰大鼠心肌组织进行泛素化修饰蛋白质组学研究，提示优化新生脉散方可进一步参与能量代谢及钙离子调控，可多途径改善心衰大鼠的运动耐量，揭示了优化新生脉散方提高心衰大鼠运动耐量效应的非心源机制。