壮医针挑疗法治疗哮喘的机制研究

Asthma is an airway allergic disease characterized by Th2-dominated （T helper cell 2-dominated）cell differentiation immune response through DCs（dendritic cells） mediating. TSLP(thymic stromal lymphopoietin) participates in regulation of Th2-dominated cell differentiation by way of DCs and some recent studies have shown that miRNAs are involved in Th2-dominated cell differentiation,furthermore,researches had shown that TSLP was regarded as a target of miRNA-126,and our preliminary researches have shown that acupuncture therapy of Zhuang nationality medicine is able to down-regulate expression of miRNA-126 ,reduce the levels of TSLP and Th2 cytokines so that to suppress inflammation reaction of asthma, therefore,we hypothesize that acupuncture therapy of Zhuang nationality medicine can firstly regulate miRNA-126, then mediate TSLP through miRNA-126, and finally regulate Th2-dominated cell differentiation and the levels of Th2 cytokines through TSLP to take an effect on the treatment of asthma . Based on our preliminary researches, in this project , we will take advantage of animal trial through acupuncture therapy of Zhuang nationality medicine to confirm our hypothesis from the levels of both gene and molecule. Our findings are expected to clarify the mechanism of the treatment of asthma by applying acupuncture therapy of Zhuang nationality medicine and provide new ideas for the targeted therapy of asthma.

哮喘是以树突状细胞（dendritic cells，DCs）介导的辅助性T细胞2（T helper cell 2,Th2）优势分化免疫应答为特征的气道变应性疾病。胸腺基质淋巴细胞生成素(thymic stromal lymphopoietin, TSLP)可通过DCs参与调控Th2优势分化的细胞因子水平，而微小RNA（miRNA）也参与调控Th2优势分化。近年研究表明TSLP 是miRNA-126的作用靶向，我们前期工作显示壮医针挑疗法能下调miRNA-126表达，降低TSLP和Th2型细胞因子水平而抑制哮喘炎症反应，推测针挑疗法通过调节miRNA-126，继而经由miRNA-126靶向调节TSLP进而调控Th2优势分化的细胞因子水平从而起到治疗哮喘的作用。本课题在此基础上，应用针挑疗法作为干预措施，通过动物实验，从基因和分子水平上探讨针挑疗法治疗哮喘的机制并为哮喘的靶向治疗提供新思路。

哮喘危害人类健康并给社会造成了沉重的医疗负担，寻求效验、低廉的治疗措施并阐明其起效机制需要研究。针挑疗法对哮喘有一定疗效，但其起效机制仍不明确。本项目建立哮喘动物模型，采用针挑疗法干预，检测微小RNA-126（miRNA-126）、胸腺基质淋巴细胞生成素(TSLP) 表达及辅助性T细胞2（Th2）型细胞因子水平，从基因和分子水平上阐明针挑疗法治疗哮喘的机制。研究结果表明针挑疗法组（针挑组）TSLP蛋白表达（62.11±20.23）、IL-4水平（18.23±1.26）、IL-5水平（130.01±28.65）、IL-10水平（96.26±33.16）、IL-13水平（113.11±31.23）、IgE水平（23.36±5.84）均降低而IL-12水平（43.05±18.12）、IFN-γ水平（32.01±1.62）升高；T-bet基因相对表达（2.63±0.73）升高而miRNA-126相对表达（2.38±0.25）、TSLP基因相对表达（4.02±0.55）、GATA-3基因相对表达（2.16±0.44）、OX40L基因相对表达（40.26±14.21）均降低；0.1秒用力呼气容积（FEV0.1）（0.62±0.06）、0.2秒用力呼气容积（FEV0.2）（1.08±0.07）、第0.4秒用力呼气容积占用力肺活量比值（FEV0.4/FVC）（0.63±0.05）、呼气峰流速（PEF）（8.05±0.28）均升高。与哮喘模型组（模型组）比较，差异均有统计学意义（P<0.05 或P<0.01）。提示miRNA-126 通过调节TSLP 从而间接调控Th2型细胞因子水平参与哮喘的调控，针挑疗法通过调节miRNA-126，继而经由miRNA-126靶向调节TSLP进而调控Th2优势分化的细胞因子水平从而起到治疗哮喘的作用。项目研究为针挑疗法治疗哮喘的临床应用提供了基础研究依据。