基于βAR-cAMP-PKA/Epac信号串扰的人参配伍环境对附子贝母反药组合减毒存（增）效的化学及生物学机制研究

Incompatibility could stimulate special efficacy when treatment of the disease, the symptom conditions and compatibility environment are key factors on the toxicity and efficacy performance. Our previous study found that Fuzi Beimu incompatibility (FBI) could persistently increase myocardial contractility but aggravate the process of heart failure (HF). Ginseng is commonly used for treatment of heart failure with Fuzi incompatibility. Whether and how Ginseng intervened with FBI to decrease toxicity and keep (or increase) efficacy in treatment of heat failure is worthy to study. Based on the previous study, we propose the hypothesis that “Increasing myocardial contractility and myocardial apoptosis are the toxicity and efficacy of FBI in treatment HF with symptom conditions. Ginseng could decrease toxicity and keep (or increase) efficacy based on βAR-cAMP-PKA/Epac signals crosstalk when FBI treat HF”. Firstly, rat MCT-induced HF models would be employed to determine the contraindication of FBI. The decreasing toxicity and keeping (or increasing) efficacy effects of ginseng combined with FBI in treatment of HF under the contraindication. To clearify the characteristics of ginseng compatibility environment on attenuating toxicity and keeping efficacy. The chemical fundamental and biological mechanism of detoxicity and efficacy saving will be interpreted by using the chemicobiology and molecular biology technology based on cAMP-PKA/Epac signal crosstalk. It will provide a new method of decreasing toxicity and keeping (or increasing) efficacy reaerach of the incompatibility, and provide new strategy for the research and development of new TCM drug based on the “Incompatibility stimulates success” theory.

配伍禁忌中药存在相反之性激其成功的特殊规律，病症条件及配伍环境是影响反药组合毒效表征的关键因素。前期发现附子贝母反药组合具有正性肌力作用并加重心衰是其毒效表现，人参常见于附子反药组合的配伍环境，其如何调控相反相激特性发挥减毒存（增）效的作用值得深入研究。基于此我们提出“增加心肌收缩力、促进心肌细胞凋亡是附子贝母反药组合干预不同病症条件心衰的毒、效表现，人参配伍环境可基于βAR-cAMP-PKA/Epac信号串扰发挥减毒存（增）效的作用”的工作假说。复制肺心病模型，进行附子贝母反药组合对的毒效表征研究，在禁忌条件下考察人参对附子贝母反药组合减毒存（增）效的作用，利用计算化学、分子生物学等方法阐释人参配伍环境对附子贝母反药组合干预心衰减毒存（增）效的化学及生物学基础。为中药配伍减毒存（增）效减研究提供新的思路与方法，为基于相反相激理论创新中药研发提供新的策略。

本课题组在前期研究基础上，认识到附子反药组合在效应发挥过程中存在激发特殊功效的特点，具有相反之性激其成功的特殊规律，功效发挥过程中产生的严重不良反应是其“毒”性的重要表现，病症条件是影响反药组合毒效表征的重要因素。历代医家在特定疾病条件下遣方用药，整理归纳了中药之间的相互作用规律，发现配伍环境是决定药物功效发挥方向（正性？负性？）的主要因素，通过合理的配伍，组合成有针对性的配伍环境，能起到减毒存（增）效的妙用。在历代反药同方配伍方剂中人参较常用于附子反药组合的配伍环境。.本研究采用腹腔注射野百合碱的方法复制大鼠肺心病模型，考察人参配伍对附子贝母反药干预肺心病心衰阶段的减毒存（增）效的作用，并深入探讨其作用机制及药效物质基础，为临床用药提供依据。研究结果表明，人参配伍环境对附子贝母反药组合加重肺心病心衰的现象具有改善作用，表现为减毒存（增）效，减毒：死亡率降低，心肌凋亡率降低，心肌酶LDH、CK、CK-MB明显降低；存（增）效：显著升高心脏射血分数、短轴缩短率，改善右心室压力负荷及心脏病理。作用机制可能与cAMP-PKA/Epac信号串扰有关，通过抑制β2AR-Gs-PKA/CaMKII信号通路，同时又激活抗凋亡信号通路Epac/Rap1/Rac/ERK1/2，减少心肌细胞凋亡，而发挥存（增）效减毒的作用。效应物质研究表明人参中发挥效应的物质可能是人参皂苷类成分。.本研究还证实了人参配伍环境可降低附子半夏反药组合干预肺心病心衰阶段的心脏毒性，并改善心功能，其作用机制可能与调节肥大、凋亡基因表达有关。