基于NO-cGMP-PKG信号通路对VSMC舒张的调节作用探索“按之则热气至”的靶器官效应机制

推拿手法作用机制一直是推拿领域里研究的热点和难点，按法是最基本手法之一，大部分手法都含有按压的动作，故按法是手法机制研究的重点手法。内经中有“按之则热气至”之说，以按压心俞穴产生热气至缓解寒凝气滞血瘀病机的冠心病心绞痛来阐释按法的温经散寒行气活血的作用。其病机的关键是冠脉痉挛，心肌缺血缺氧。诱导冠脉平滑肌舒张可有效的缓解心肌缺血缺氧，NO-cGMP-PKG信号通路在调节血管舒张方面有重要作用。本项目以此为切入点，以寒凝气滞血瘀型冠心病大鼠模型为受试对象，运用激光多普勒血流分析、红外测温技术、免疫组化、Western-blot 等技术，通过按压心俞穴，观察穴位局部温度、心肌缺血后心电图、心脏微循环和组织形态的变化，以及VSMC胞浆内NO、cGMP、PKG含量变化和NO-cGMP-PKG信号通路下游环节中各调节蛋白含量或磷酸化状态及相关的变化，探索“按之热气至”的靶器官效应可能机制。

The mechanism of “Tui-na” Manipulation is hotspot and difficulty research in this field, and Pressing Manipulation is the key of research, because Pressing Manipulation is one of the most basic manipulation, and most manipulations contain press action. The yellow emperor's Canon has the say "After pressing, It Heat Up" to interpretation Promoting Flow of Qi & Blood and Warming Channel for Dispelling Cold effect of Pressing Manipulation, and use the case of CHD Angina Pectoris of Congealing Cold with Blood Stasis and Qi Stagnation pathogenesis was relieve through Acupressing Shenshu Point produces heat. The key factor for its pathogenesis is Coronary Spasm, myocardial ischemia hypoxia. Induced coronary artery smooth muscle relaxation can effectively alleviate myocardial ischemia hypoxia, the NO-cGMP-PKG signaling pathways play an important role in regulating vasodilation. This project take this as the breakthrough point to explore the possible mechanism of target organ effect of "After pressing, It Heat Up". As the Model of Congealing Cold with Blood Stasis and Qi Stagnation CHD Rat as the subjects. And using the laser doppler blood flow analysis, infrared temperature measurement technology, immunohistochemistry and Western blot techniques. And acupressing the Shenshu point to observation the changes of points local temperature, electrocardiogram, heart microcirculation after myocardial ischemia, and organization form. And the changes of VSMC intracytoplasmic NO, cGMP, PKG content. As well as the changes of NO-cGMP-PKG signaling pathways downstream link regulatory proteins or phosphorylation status.