从Sig-1R调控内质网/线粒体信号转导角度探讨益气活血法干预心衰心肌细胞损伤的机制研究

Subcellular organelle injury and connection disorders of endoplasmic reticulum and mitochondria in the stress state is a new molecular mechanism of the heart failure (HF). Sigma-1 receptor (Sig-1R) regulated calcium transport and stress induced signaling transport between endoplasmic reticulum and mitochondria determines the final outcome of the endoplasmic reticulum stress and mitochondrial stress. Activation of the Sig-1R regulating calcium transport between endoplasmic reticulum and mitochondria is a new field of research on the prevention and control of HF. Traditional Chinese medicine holds that the pathogenesis of HF is deficiency of Qi and blood stasis, supplementing qi and activating blood circulation method can obviously improve the cardiac function. This project mainly uses the rat HF model by coronary artery ligation system, combined with AngII induced hypertrophy model of primary cultured myocardial cell, by echocardiography, tissue pathology, real-time PCR, Western blot and RNA interference methods to to investigate the pathological change of HF and traditional Chinese medicine Yiqihuoxue compound mechanism. In order to demonstrate that Yiqi Huoxue Prescription treat HF by adjusting the chaperone Sig-1R, regulating of endoplasmic reticulum and mitochondrial calcium transport, interventing of endoplasmic reticulum and mitochondria stress injury, improving myocardial energy metabolism, so inhibiting myocardial cell injury. Our project from the microcosmic angle, provide the experimental basis for the clinical application of " scientific and objective the pathogenesis theory of qi deficiency and blood stasis " in the prevention and treatment of HF.

内质网/线粒体亚细胞器损伤及联系障碍是导致心力衰竭（HF）的新分子机制。作为调节内质网/线粒体信号联系的关键因子，Sigma-1受体的激活对调控内质网/线粒体钙转运及应激损伤信号通路，防治HF具有重要意义。气虚血瘀是HF的基本病机，以益气活血法为指导思想的益气活血方可明显改善心肌能量代谢，防治心梗后HF。推测其作用机制可能通过激活Sig-1R调控内质网/线粒体钙转运，干预内质网/线粒体应激损伤，改善能量代谢，抑制心肌细胞损伤实现的。在此基础上，我们采用冠状动脉结扎术制备大鼠心梗后HF模型，结合AngII诱导的原代培养的心肌细胞肥大模型，通过心脏超声、组织病理、定量PCR、Western杂交以及小RNA干扰等方法，在亚细胞器分子水平，探讨HF过程中细胞能量代谢障碍、凋亡的发生机制及益气活血中药复方的干预作用。拟从微观辨证角度，为“气虚血瘀”病机理论在防治HF的临床应用提供科学客观的实验基础。

益气活血法治疗心力衰竭可以改善心肌能量代谢障碍。Sigma-1受体作为调节内质网/线粒体信号联系的关键因子，参与调控内质网/线粒体钙转运，影响能量代谢，保护心肌细胞功能。本研究通过在体实验验证了心力衰竭心脏组织能量代谢障碍。证实益气活血药可以改善心肌组织的结构损伤和能量代谢障碍。在分子水平检测结果提示益气活血药的这些作用与干预内质网应激信号通路和sigma-1受体的表达相关。在细胞水平，研究证实，益气活血药活性成分单体磷酸川芎嗪可以通过干预sigma-1受体的表达及内质网应激信号通路影响细胞内钙平衡，抑制细胞肥大。.这些结果提示sigma-1受体可能作为心力衰竭治疗的新靶点，同时为益气活血中药治疗心力衰竭提供新的实验依据。