基于"穴位-鼻黏膜感觉神经-树突状细胞"偶联的穴位埋线治疗变应性鼻炎的作用机制研究

This study is based on our former project of the National Natural Science Foundation 〝 the regulatory effects of catgut implantation at acupoints on neurogenic inflammation via axon reflex in allergic rhinitis (AR)〞. In our previous studies, we found that catgut implantation at acupoints could inhibit the release of neuropeptides in the nasal mucosa, such as SP(Substance P), VIP(Vasoactive Intestinal Peptide), CGRP(Calcitonin Gene-related Peptide), and then the neurogenic inflammation of the nasal mucosa was relieve. However, the mechanism of how catgut implantation at acupoints regulates the allergen-derived inflammatory remains unclear. In order to further explore the mechanism, we propose a hypothesis of the coupling of 'neuropeptide -sensory nerve of nasal mucosa - dendritic cell (DC)',which means the change of the neuropeptides in the nasal mucosa could regulate the activity of antigen-presenting cell(DC) in different stages of mucosal immune function. First,by the methods of histology and molecular biology techniques, immature DCs (iDC) are isolated and cultured in vitro, then co-cultured with CD4 effector T cells and treated with exogenous neuropeptides. The differentiation of DC, the process of antigen presentation and the differentiation of effector T cell would be studied.Second,allogeneic exogenous DCs, which have been treated with neuropeptides and fluorescently labeled, would be re-implanted again in the nasal mucosa of the rat. The density, distribution and differentiation of iDC would be studied. Finally, the rat models of AR are divided into the study group and control group. The study group would be treated with catgut implantation at Yingxiang. The change of Claduin-1, Toll-like receptors, cytokines would be studied by western blot and ELESA methods.And the coupling cytomorphology of DC and sensory nerve will be observed by light and electron microscope. We hope we could clarify the mechanism of how catgut implantation at Acupoints could regulate the allergen-derived inflammation of AR.

本研究是国家自然基金课题"穴位埋线经轴索反射对变应性鼻炎（AR）神经源性炎症的调控机制"的深化研究。前述研究发现穴位埋线通过抑制鼻黏膜P物质等感觉神经肽的释放减轻神经源性炎症从而缓解AR症状。但穴位埋线如何最终实现对AR的调节机制不清。本研究提出"穴位-鼻黏膜感觉神经-树突状细胞（DC）"偶联假说，穴位埋线通过抑制鼻黏膜感觉神经肽的释放进而调节抗原呈递细胞（APC）-DC对鼻黏膜免疫反应的影响。采用组织学、分子生物学等技术，分别通过DC与CD4效应T细胞体外联合培养的离体实验、经感觉神经肽预处理的同种异体外源性DC重新植入大鼠鼻黏膜内的在体实验，观察感觉神经肽对DC分化、抗原呈递及Th分化的影响。制作AR大鼠模型、行穴位埋线，对鼻黏膜组织学、感觉神经与DC的偶联形态学等行光镜与电镜观察，检测感觉神经肽、Claduin-1、Toll样受体、有关细胞因子等，进一步阐明穴位埋线治疗AR的作用机制

2014年自然基金面上项目——基于"穴位-鼻黏膜感觉神经-树突状细胞"偶联的穴位埋线治疗变应性鼻炎的作用机制研究（编号81473523）。该项目于2015年1月启动，于2018年12月截止，按时标书设计共分为三个研究阶段：第一阶段，对iDC进行体外分离培养，与CD4效应T细胞进行联合培养，在经外源性神经肽处理后，对DC的分化、抗原呈递及效应T细胞Th分化的影响进行研究；第二阶段，通过神经肽处理后的同种异体外源性DC重新植入大鼠鼻腔黏膜内的再培养研究；第三阶段，对AR 模型行迎香穴埋线，对Claduin-1、Toll样受体、细胞因子等进行检测，阐明穴位埋线的机制。在该项目执行过程中，我们严格遵循标书设计要求，保质保量的完成阶段研究，客观分析各项研究结果，并针对课题设计环节中的不足进行及时校正。目前，该课题各项主要研究目标，已顺利完成。依托该课题，发表SCI 5篇，中文核心期刊论文19篇，进行全国性会议交流8次，建立科普微信平台1座，培养硕博士研究生共16人。并以本次课题为基础，成功申请2016年、2017年国家自然科学基金项目2项，进行该课题的深化研究。综上，本次课题完成情况已达到标书预期，进行结题，并将结果向国家呈报。