穴位埋线经"SP-ILC2"信息联动对局部变应性鼻炎的调节机制

Local allergic rhinitis (LAR) is an independent disease entity which coexists with classical allergic rhinitis. A specific feature of LAR is that there is only local nasal mucosal immune response with absence of systemic anaphylaxis indexes. Our previous studies showed catgut implantation at acupoints is effective for LAR, but the mechanism was not clarified. Type II innate lymphoid cells (ILC2s) have a potential role in the immune response of LAR. Our previous research also suggested a close mathematical correlation between neuropeptide substance P (SP) and ILC2. Therefore, catgut implantation could effectively regulate the expression of SP in the nasal mucosa. We speculated that “SP-ILC2” information linkage might be the necessary gateway for implementing the regulation of catgut implantation for LAR by inhibiting the expression of SP and inactivating ILC2, thereby inhibiting the downstream immune response. As a result, the symptoms of LAR would be alleviated. Based on this hypothesis, we plan to conduct our studies in three stages: first, the in vitro verification of “SP-ILC2” information linkage would be done by stimulating ILC2 with SP of a known concentration gradient; second, the LAR animal model would be established to verify the “SP-ILC2” information linkage in LAR immune microenvironment by inhibiting SP and ILC2 with capsaicin and monoclonal antibody respectively; and third, the regulatory effect of acupoint catgut embedding on LAR through the “SP-ILC2” information linkage will be observed. By this, the mechanism of acupoint catgut implantation therapy in improving LAR would be clarified in a methodical manner.

局部变应性鼻炎（LAR）是仅有鼻腔局部过敏反应而全身性过敏指标阴性的局部黏膜免疫疾病。我们前期发现穴位埋线能有效改善LAR但机制不清。研究进展提示II型固有淋巴细胞（ILC2）是LAR免疫应答启动及推进核心；我们还发现感觉神经肽P物质（SP）与ILC2间存在线性联动关系，且穴位埋线调节SP作用显著，因此推断“SP-ILC2”信息联动是穴位埋线调节LAR的必经环节，即：穴位埋线下调SP，降低ILC2活性，抑制免疫进程，恢复LAR免疫稳态。为此，我们首先行“SP-ILC2”信息联动体外验证，以浓度梯度SP刺激ILC2，建立二者联动变化量效关系；其次，建立LAR动物模型，分别以辣椒素、ILC2单克隆抗体抑制SP、ILC2活性，在LAR黏膜免疫微环境中动态验证“SP-ILC2”信息链；最后，对LAR动物实施穴位埋线，通过SP、ILC2的关联性分析，最终阐明穴位埋线对LAR的调节机制。

局部变应性鼻炎（LAR）是仅有鼻腔局部过敏反应而全身性过敏指标阴性的局部黏膜免疫疾病。我们前期发现穴位埋线能有效改善LAR但机制不清。研究进展提示II型固有淋巴细胞（ILC2）是LAR免疫应答启动及推进核心；我们前期研究发现穴位局部埋线刺激感觉神经产生信息调节物—感觉神经肽P物质（SP），我们还发现SP与ILC2关联密切。因此推断：穴位埋线经“SP-ILC2信息联动”调节LAR，即：穴位埋线下调鼻黏膜SP，降低ILC2活性，改善LAR免疫进程，恢复鼻黏膜免疫稳态。对此假说我们分三步走予以验证。第一步为体外试验，旨在细胞学水平验证“SP-ILC2信息联动”。为此，首先分离富集并原代培养ILC2后以梯度浓度SP刺激ILC2，建立二者联动变化量效关系；其次动态检测“SP-ILC2量效体系”中ILC2转录因子 GATA-3、抑制性受体 IFN-γ、细胞因子 IL-5、IL-13，进一步解析经SP刺激后ILC2上、下游关键细胞因子的动态变化；然后筛选“SP-ILC2”联动体系差异化表达基因，剖析基因功能及信号通路指向，进一步阐明“SP-ILC2”联动的分子生物学基础。第二步为体内试验，旨在动物学水平验证“SP-ILC2信息联动”量效关系。为此，首先建立LAR动物模型；其次检测并分析鼻黏膜SP与ILC2细胞因子（IL-4、IL-5、IL-13）、ILC2标志化基因（CD90、CD117、CD127、ST2、Thy）的动态关联变化；然后以辣椒素阻断SP后再次分析“SP-ILC2”关联变化。第三步为加入穴位埋线刺激因素后的体内试验，旨在动物学水平验证穴位埋线经“SP-ILC2信息联动”对LAR的调节作用。为此建立LAR动物模型后，首先予以不同干预措施（穴位埋线/非埋线治疗），同时设非致敏空白对照组；然后检测并分析鼻黏膜SP与ILC2细胞因子、ILC2标志化基因的关联物变化。通过以上“三步走”方案，我们不仅证实ILC2为LAR核心免疫细胞，而且从细胞到动物在不同维度逐步建立并验证了“SP-ILC2信息联动”体系，然后逐层递进最终证实穴位埋线经“SP-ILC2信息联动”调节LAR的机制假说。