鼻敏煎调节γδT细胞重塑Th1/Th2/Treg/Th17治疗变应性鼻炎的免疫机制

The main pathogenesis of allergic rhinitis is the immunologic dissonance of Thl/Th2/Treg/Th17. Allergic rhinitis is one of the dominant diseases which can be treated with TCM. Using BIMINJIAN Syrup to cure allergic rhinitis has been shown with great therapeutic effect. Previous studies have shown that BIMINJIAN Syrup could regulate the central memory T cells and the central effector T cells and affect the expression of Th1/Th2 cytokines in patients with allergic rhinitis. However, the mechanisms by which BIMINJIAN Syrup plays a role in invigorating qi for consolidating superficies, changing the local mucosal immune inflammatory status, controlling the clinical symptoms, and regulating the immune status remains unknown. Based on the characteristics of the γδT cells which are widely distributed in the mucosal tissues and the regulating function which is independent on antigen processing and presentation of the innate and adaptive immune regulation, we designed the in vivo experiment with the mouse models of AR and observed the gamma delta T cell and the Th1/Th2/Treg/Th17 immune balance in the blood and nasal mucosa after treatment with BIMINJIAN Syrup . We also designed the in vitro experiment with medicated serum of BIMINJIAN to investigate the influence of BIMINJIAN Syrup on the activation and proliferation of the γδT cells , and the cellular signaling pathway activated by the γδT cells. We seek to clarify the mechanism by which γδT cells remodel Th1/Th2/Treg/Th17 immune imbalance. These findings also have important implications for finding new targets in the treatment of allergic rhinitis by Chinese medicine, as well as realistic significance in exploring the mechanisms of TCM treatment of allergic rhinitis and promoting the TCM treatment of allergic rhinitis in the world.

Thl/Th2/Treg/Th17免疫失衡是变应性鼻炎(AR)的主要发病机制。验方“鼻敏煎”治疗AR疗效显著，前期研究表明鼻敏煎能调节AR患者Tem和Tcm，影响Th1/Th2免疫平衡，但鼻敏煎的“益气固表、扶正固本”作用如何改变黏膜局部免疫炎性状态，控制临床症状的作用机制还不清楚。鉴于γδT细胞在粘膜组织优势分布以及不依赖抗原处理和呈递的固有免疫和适应性免疫调节作用，本课题通过体内实验研究AR小鼠模型经鼻敏煎治疗后，血液及鼻粘膜γδT细胞、Th1/Th2/Treg/Thl7细胞因子的表达，阐释鼻敏煎对γδT细胞的调节作用及对Th1/Th2/Treg/Thl7免疫失衡的影响。通过体外实验研究鼻敏煎含药血清对γδT细胞活化和增殖的影响，以及γδT细胞的活化信号传导通路，明确γδT细胞重塑Th1/Th2/Treg/Thl7免疫失衡的作用机制。本研究对中药治疗AR新作用靶点的发现具有重要意义。

背景：鼻敏煎是治疗变应性鼻炎（AR）的验方，疗效显著，前期研究发现鼻敏煎可调节AR外周血Th1/Th2/Treg/Th17免疫失衡，但其机制有待探究。γδ T细胞是气道黏膜免疫重要的功能细胞，对固有免疫和适应性免疫发挥重要的调节作用，研究鼻敏煎干预γδ T细胞对Th1/Th2/Treg/Th17免疫平衡的调节作用及其机制对于揭示鼻敏煎的免疫治疗作用具有重要意义。.研究内容：1. 研究鼻敏煎对AR小鼠干预作用。2. 体内实验研究鼻敏煎对γδ T细胞调节作用及对Th1/Th2/Treg/Th17免疫失衡的影响，明确其免疫机制。鼻敏煎干预AR小鼠模型，观察小鼠外周血、鼻黏膜γδ T、Th1/Th2/Treg/Th17细胞及其相关因子的表达。3. 体外实验研究鼻敏煎对γδ T细胞活化和增殖作用，及对γδ T细胞活化信号传导通路的影响，明确γδ T细胞重塑Th1/Th2/Treg/Th17免疫平衡的作用机制。含药血清处理γδ T细胞，检测Vδ1、Vδ2 T细胞活化后Th1/Th2/Treg/Th17细胞因子、信号传导通路磷酸化、核内转录因子T-bet、Gata3、Foxp3和RORγt的水平。.重要结果及关键数据：1. 鼻敏煎对AR小鼠有干预作用。2. 鼻敏煎显著抑制Th2、Th17细胞（P＜0.05），细胞因子IL-2、IL-4、IL-5、IL-17及TGF-β水平低（P＜0.01或0.05），IL-10高表达（P＜0.01）；鼻敏煎明显减少小鼠鼻黏膜嗜酸性粒细胞浸润（P＜0.05）；鼻敏煎下调转录因子GATA3、RORγt、γδ TCR表达（P＜0.05），其相应mRNA表达下调（P＜0.05或P＜0.01）；3. 鼻敏煎含药血清抑制Vδ1 T细胞STAT3磷酸化（P<0.05），抑制Vδ1、Vδ2 T细胞中PLC-γ1磷酸化，降低细胞因子IL-4、IL-6、TNF-α水平，升高IL-10水平（P<0.05），降低γδ T细胞T-bet表达、升高GATA3表达（P<0.05）。 .科学意义：鼻敏煎通过干预γδ T细胞重塑Th1/Th2/Th17/Treg免疫失衡从而起到祛邪扶正、调整机体免疫功能的作用；信号通路STAT3、PLC-γ1以及核内转录因子T-bet、GATA3作为鼻敏煎活化γδ T细胞的可能作用靶点，为中医药治疗AR提供了新思路。