不同强度针灸诱发穴区IL-17A免疫反应与心肌NLRP3炎症小体活性的相关性研究

During the study of cardioprotection of acupuncture preconditioning on myocardial ischemia reperfusion injury (MIRI), we found that the effect was related to the expression of inflammative substances in “neiguan”acupoint. Simutaneously, our RNS-seq results showed that the immune and metabolism pathways varied prominently in normal people treated by acupuncture. Additionally, recent research shows that nociceptive neurons could promote innate immune response via interleukin-17A (IL-17A) from γδT cells drived by IL-23 of dermal dendritic cells. According to the above evidences, we put forward the following hypotheses: local IL-17A immune response induced by suitable acu-moxibustion stimulation is the key mechanism for disease prevention. In light of the hypothesis, heart function will be protected from MIRI by preconditioning wth different acupoints with different intensity of acu-moxibustion pretreatment depend on the theory of the meridian viscera correlation. Many experimental techniques and methods would be employed in the study, such as target protein in vivo blocking technology, flow cytometry, qPCR, Western Blot and so on. After evaluation of therapeutic effects, we would confirm the relationship between the effective activition of IL-17A immune response in acupoints and MIRI protection accroding with the following route”acupuncture with different intensity—activate IL17A immune response—mediate holistic immune regulation—induce myocardial NLRP3 inflammasome’s activity changes—achieve cardioprotection effect” cascade, combined with the correlational analysis. Meanwhile, this study will also uncover the mechanism of local IL-17A-mediated immune response induced by nociceptive sensory neuron-immune interaction. Further more, we will investigate the scientific connotation of acupoints choosen, needle methods and acupuncture intensity, and provide scientific evidence and guidance to acupuncture’s clinical application in disease prevention.

我们针灸预处理抗心肌缺血再灌注损伤（MIRI）研究发现，穴区炎症免疫物质表达与效应存在相关性；测序结果显示：正常人针刺前后免疫代谢信号通路变化明显；结合免疫防御的现代认识及伤害感受神经元诱导局部IL17A免疫反应的新前沿，提出“适宜针灸刺激诱发局部IL17A免疫反应是实现治未病的关键启动机制”假说，在经穴-脏腑相关理论指导下，选用不同穴位给予不同强度针灸预处理后，制作MIRI模型。采用靶标在体阻断技术结合流式细胞术、qPCR、WB等实验技术，在评价心肌保护效应后，围绕“不同强度针灸—启动穴区IL17A免疫反应—整体免疫响应—调控心肌NLRP3炎症小体活性—心肌保护”效应级联展开研究，结合关联分析，明确有效启动穴区IL17A免疫反应是实现MIRI保护的关键，揭示诱发该局部免疫反应的伤害感受神经元-免疫交互调控机制，探寻该过程穴位、针灸方法及强度选择的科学内涵，为针灸治未病的科学应用提供依据。

治未病健康理念持续深入，针灸养生保健以治未病正被健康爱好者所青睐，深入揭示其作用规律和机制对促进临床科学应用具有积极意义。腧穴、刺激方法和刺激强度选择是实现针灸临床效应的核心内容，但目前缺乏统一指导原则。内关是治疗心肌损伤最常用腧穴，电针是常用的刺激方法，而刺激强度选择则存在很大差异，那么腧穴局部刺激强度、局部炎性反应、机体免疫反应、靶器官效应之前存在什么关系？本研究就此进行探索。.基于免疫系统防御、监视、自稳功能的认识和“心胸内关谋”经典理论，本研究沿用“内关”穴进行电针干预，以心肌损伤模型鼠为研究对象，结合伤害感受神经元诱导局部IL-17A免疫反应的新前沿，在充分评价不同强度电针产生心肌保护效应的基础上，探索穴区炎症与机体免疫应答和心脏组织NLRP3炎性小体活化及心脏功能之间的关系，明确局部IL-17A的心肌保护效应，并揭示其作用机制。结果表明，内关穴的适宜刺激（小鼠1mA，大鼠2mA）是实现最佳心肌保护效应的基础，其能有效激活TRPV1、CGRP信号、启动加强穴区组织中IL-17A为代表炎性反应强度，调节机体免疫反应，抑制损伤心肌组织中NLRP3炎性小体活化，促进巨噬细胞M2型极化，减少IL-6、IL-1β、TNF-α水平，减轻炎性反应，减小心肌梗死面积，提高左心室射血分数和短轴收缩率，实现心肌保护效应。并采用穴位注射IL-17A或CFA以诱发穴区炎症和手术切断迷走神经及摘除脾脏的方式，进一步探索穴位局部炎症与靶器官保护及自主神经抗炎免疫途径在该过程的作用，结果表明，穴区适度炎性反应是产生心肌保护效应基础，迷走神经-脾轴或心-脾轴参与该过程。同时，初步揭示穴区诱发炎性反应或电针可通过调控β1β2肾上腺素能受体信号通路、改善肠道菌群丰度、调节损伤心肌局部自主神经重塑实现促心肌保护效应。通过本研究，揭示电针预处理诱发炎症免疫反应调控心肌组织炎症反应介导心肌保护效应机制，并初步提出“以炎抗炎”的针灸治未病思路。该结果对电针治未病临床实践具有指导意义，也为后续开展不同部位腧穴效应特点研究奠定基础。