基于Sirtuins/AMPK-PGC-1α通路研究活血化瘀复方丹七片调控缺血心肌能量代谢紊乱的机制

Coronary heart disease is the leading cause of death in the world. Coronary heart disease is caused by atherosclerosis and the blockade of heart arteries will lead to short supply of oxygen and nutrients to the heart cells. Energy metabolism disorder in mitochondria is the basis for pathological development in the ischemic heart. Treatment with Traditional Chinese medicine Fufang Danqi pill could effectively alleviate the symptoms of coronary heart disease. Our previous study showed that Danqi pill could regulate fatty acids metabolism disorders in ischemic heart. However, the molecular mechanism of Danqi pill on the regulation of energy metabolism is unknown. There are several hundreds of chemical constituents in Danqi and the effective constituents are yet to be identified. In this study, we aim to investigate the regulative mechanism of Danqi pill on AMPK/Sirtuins-PGC-1α-PPARα/ERRα/NRF pathway which plays a key role in energy metabolism. Rat model with ischemic heart and hypoxic cell model will be applied in this study. We also aim to investigate the effective chemical components in Danqi pill by network pharmacology methods and evaluate their effects by experimenting on rats and cell models. This study will further our understanding of the pharmacological mechanism of Danqi pill, lay the foundation for development of new drugs targeting mitochondria and provide theoretical evidence for the treatment of other ischemia diseases by Danqi pill.

冠心病严重危害人类健康。心肌缺血导致的线粒体能量代谢紊乱是其重要的发病基础。中药复方丹七片在冠心病治疗中有确切的疗效。我们前期研究表明，丹七片能够改善缺血心肌的脂肪酸代谢紊乱，但其药效机制尤其是对整体能量代谢的调节机制尚不清楚，且其发挥药效的有效组分尚未阐明。本研究将以心肌缺血大鼠为动物模型，并建立体外心肌细胞缺氧模型，采用Western blot、定量PCR等技术，从分子水平上研究丹七片对以PGC-1α为核心的Sirtuins/AMPK-PGC-1α线粒体能量代谢通路的调节机制；采用液相色谱/串联质谱联用方法，分析药物入血成分；采用网络药理学方法，分析丹七片中作用于能量代谢网络的起效成分，并对起效成分进行生物学验证，从分子网络水平阐明复方丹七片治疗缺血性冠心病中的药理机制。该研究将进一步深化对中药复方的药效机制认识，为以线粒体为靶标的新药开发提供理论基础，并为丹七片治疗其它缺血性疾病提供理论支持。

冠心病是冠状动脉供血不足导致的严重危害人类健康的疾病，心肌缺血导致的线粒体能量代谢紊乱是其重要的发病基础，中药复方丹七片在治疗冠心病方面有确切疗效，但其对整体能量代谢的调节机制尚不清楚。本课题通过建立心肌缺血大鼠动物模型和心肌细胞缺氧模型，系统研究了丹七片对以PGC-1α为核心的Sirtuins/AMPK-PGC-1α线粒体能量代谢通路的调节机制。该研究发现，丹七片能够保护模型大鼠的心功能，激活Sirtuins/AMPK-PGC-1α通路，并上调下游靶标中Mfn1和SOD2的表达，发挥激活线粒体活性以及抗氧化功能。细胞实验结果表明，丹七片能够显著提高H9C2细胞模型中ATP的含量，并能显著降低活性氧基团ROS的含量。细胞实验也进一步证实，丹七片可提高PGC-1α, AMPKα和Sirtuin1的表达，并提高Mfn1和Mfn2的表达。该研究将进一步深化对中药复方的药效机制认识，为以线粒体为靶标的新药开发提供理论基础，并为丹七片治疗其它缺血性疾病提供理论支持。