参附配伍通过PGC-1α/ERR改善心力衰竭线粒体能量代谢重构

Mitochondrial function played important roles in metabolic remodeling of cardiac failure, mitochondria was attacked by pathology factor of cardiac failure, and mitochondria was also genesis of cardiac failure. In the previous study, the function of shenfu fang involved in energy metabolism and mitochondrial function. The effects of shenfu fang on metabolic remodeling and PGC-1α-ERR pathway in heart failure rats were investigated, the effects of shenfu fang on mitochondrial structure and function were observed, the effects of shenfu fang on mitochondrial permeability transition pore, energy metabolism, activity of respiratory chain, Ca2+ overload, reactive oxygen species, apoptosis, autophagy on mitochondria were also studied. The compatibility mechanism of aconitine and saponin of red ginseng were clarified based on the PGC-1α/ERR pathway. The role of the pathway and target of shenfu fang in metabolic remodeling was identified. Screening platform based on target was established, active components of shenfu fang were discovered, new drug based on active components were studied. The data will help new herbal drugs development and renew the old drugs.

心力衰竭代谢重构以线粒体结构功能损伤为核心，线粒体既是心力衰竭时病理因子攻击的靶标，也是心力衰竭时各种病理变化的起源。前期发现参附方心血管保护作用与改善心脏能量代谢和线粒体功能有关，本课题在饮片、组分、成分三个层次，在临床、动物、细胞、分子四个水平，考察参附配伍对心力衰竭心脏能量代谢重构、线粒体结构和功能、心室重塑等指标的影响，重点研究参附配伍对心肌细胞中PGC-1α/ERR相关通路的影响，从PGC-1α/ERR通路探索乌头碱与人参皂苷配伍的减毒增效机制。明确参附方改善心肌能量代谢的信号传导通路和关键调控因子，发现参附方作用的关键生物标志物和作用靶点，基于作用靶点构建筛选体系，发现参附方改善慢性心力衰竭心脏能量代谢重构的效应组分，进行组分中药的成药性研究。课题研究为中药二次开发，组分配伍新药提供有效支撑。

心力衰竭代谢重构以线粒体结构功能损伤为核心，课题考察了参附配伍对心脏能量代谢重构、线粒体结构和功能、心室重塑等指标的影响。研究首次揭示了附子对心肌细胞线粒体产生毒性，从而引起心肌细胞线粒体功能障碍，能量代谢异常，这可能与附子产生心脏毒性的作用机制有关；研究考察了参附配伍减毒作用机制，发现附子中主要毒效成分乌头碱致心肌细胞钙紊乱的机制，以及人参中有效成分人参皂苷Re对心肌细胞的保护作用，为研究乌头碱致心律失常作用提供线索，同时为参附配伍减毒作用机制的研究提供参考；研究发现附子中乌头碱毒效转换的新途径，高剂量（1-10μM）乌头碱促进NFAT3从胞浆转移至胞核，进而引起CaMKⅡ磷酸化水平和RyR2蛋白表达水平升高，导致心肌细胞钙瞬变频率增加以及Ca2+含量升高，细胞内钙紊乱。低剂量（5-20nM）通过激活Sirt3的活性，减轻Cyp-D的乙酰化程度，进而减少Cyp-D与ANT的结合，减少MPTP的持续性开放，减轻线粒体损伤；用药剂量会影响附子“毒-效”转化，在一定剂量内附子药效与剂量存在“量-效”关系。课题研究阐明了附方改善心肌能量代谢的信号传导通路和关键调控因子，明确了参附方作用的物质基础和作用机制，为参附方组分配伍新药研究提供了有效支撑。