针灸调控伤口炎症促进愈合的机制

Cutaneous wound healing can be promoted by acupuncture and moxibustion in clinical. Previous studies have proven that the substance P (SP) plays a vital role in anti-inflammatory and wound healing. Interestingly, acupuncture and moxibustion were proved to promote wound healing through facilitating the release of SP and regulating the release of the proinflammatory and anti-inflammatory factors. Therefore, we hypothesize that acupuncture and moxibustion could accelerate the inflammatory response by stimulating the release of SP from local nerve endings of the wound and regulating the macrophages, and promote the wound healing by activating the anti-inflammatory factors and promoting the proliferation recovery. In the present study, we aimed to observe the acceleration of wound healing by acupuncture and moxibustion. We will focus on the regulation of acupuncture and moxibustion to investigate the promotion of local blood flow and formation of tissue collagen fiber after SP release using laser doppler imaging, morphology, ELISA and flow cytometry technologies. Furthermore, we will investigate the effect of acupuncture and moxibustion on local tissue monocytes, macrophage polarization ,SP and NK-1R on M1 macrophage, the conversion of M1 to M2, and the level of proinflammatory and anti-inflammatory factors. In summary, we will elucidate the mechanism of acupuncture and moxibustion in regulating the inflammatory responses and promoting wound healing from the overall, cellular and molecular levels.

临床证明针灸可明显促进皮肤创伤的愈合。研究表明神经肽P物质（SP）在抗炎和伤口愈合过程中发挥重要作用，而针灸可促进局部SP的释放并调控巨噬细胞释放促炎和抗炎因子促进愈合。因此提出“针灸可通过刺激伤口局部神经末梢释放SP调控巨噬细胞加速炎症反应，调动抗炎因子并提前进入增殖修复期以促进伤口愈合”的假说。采用皮肤组织外伤模型，观察针灸干预加快创伤修复的进程。聚焦针灸对创伤炎症期的调控，从针灸加速炎症反应和抗炎入手，观察针灸通过激活SP的释放促进炎症期创伤局部血流量和组织胶原纤维生成；分析针灸对炎症期创伤局部组织单核细胞、巨噬细胞计数分型、SP及其受体NK-1R在M1巨噬细胞的表达及M1向M2的转化，测定诱导M1向M2分化的IL-4、HO-1，M1型巨噬细胞释放促炎因子TNF-α、NO、IL-6和M2型释放抗炎因子Arg1，IL-10，从整体、细胞和分子水平阐明针灸调控炎症反应促进伤口愈合的机制。

皮肤创伤的发病率高，修复过程复杂，易造成愈合不良等问题，产生巨大的经济、社会负担，严重影响患者的生活质量。艾灸可以促进改善创伤局部微循环状态，临床上常用于褥疮等难愈性创伤的治疗。因此，本研究将从形态学和分子生物学角度探索艾灸对全层皮肤切除创伤模型愈合的效应及其机制。.选用健康成年Sprague-Dawley（SD）大鼠为研究对象，观察艾灸对创伤模型大鼠创口愈合的效应。在确定艾灸可明显促进皮肤创伤愈合的基础上，通过荧光免疫组织化学、HE和Masson染色的形态学手段，以及Elisa和液态芯片法检测蛋白含量的分子生物方法，探讨艾灸干预对创伤愈合炎症期表达曲线的影响，以及对不同阶段创伤局部组织形态的修复作用，为相关基础和临床研究提供借鉴。揭示艾灸促进伤口愈合效应的神经生物学机制。.研究结果如下：.1.艾灸干预明显促进皮肤创伤模型大鼠的创伤修复，缩短创伤愈合的进程。.2.艾灸可显著促进创伤模型大鼠血清中炎症期促炎性细胞因子IL-1α、IL-1β、IL-2和IL-6以及抗炎性细胞因子IL-4和IL-10的动态表达曲线左移，促进创伤炎症期炎症反应，并缩短炎症期促使创伤修复提前进入细胞增殖期。.3.艾灸可增加创伤模型大鼠血清中与巨噬细胞关系密切的趋化因子G-CSF、GM-CSF、M-CSF的含量，并且调控创伤局部巨噬细胞从M1向M2型转化，提示艾灸干预通过影响巨噬细胞分型促进了炎症期反应，缩短炎症期提前进入修复期，加速创伤的愈合。.4.艾灸可增加炎症期血清及创伤周围局部皮肤组织中VEGF的含量，促进血管内皮细胞CD31阳性细胞的表达，促进了创伤修复期创伤局部血管的新生和重建。.5.艾灸可以增加创伤周围组织中神经肽SP/CGRP的表达，主要表达在表皮下和血管周围，诱导神经源性炎症反应的发生，提示艾灸促进神经肽的释放，诱导下游的神经免疫反应使其快速进入修复期来促进外伤的愈合。