“冬病夏治”中药敷贴对哮喘大鼠树突状细胞免疫功能的影响

发布时间：2018-02-28 20:47

本文关键词： 冬病夏治中药敷贴哮喘树状突细胞免疫功能　出处：《成都中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：研究背景：支气管哮喘是儿童最常见的慢性呼吸系统的疾病,是由多种炎性细胞及炎性介质相互作用、互相影响,而引起的呼吸道慢性炎症性的疾病,近几年来的实验研究,有学者提出支气管哮喘反复发作的主要原因是其机体的免疫失常,而最重要的免疫异常是Thl/Th2细胞数目比例和其功能的失去平衡[1]。DC是就目前为止我们所发现的抗原提呈细胞中功能最强的免疫细胞,它除了所具有的独一无二的刺激初始T细胞增殖的能力,还能够直接干预Th0细胞的分化的方向,从而导致Thl/Th2细胞数目比例失调而引起哮喘发作[2]。“冬病夏治”穴位敷贴疗法是中药外治法中最具特色治疗方法,是外用药制剂中最具有典型性的代表方剂之一,它的机理是通过药物对经络穴位刺激和有效成分透皮渗透吸收的双重作用,来实现防治哮喘的目标[3]。本实验在结合以往对于哮喘大鼠相关的DC诱导及培养及异常指标的基础上,提出“冬病夏治”穴位敷贴疗法能通过下调树突状细胞的表达,抑制DC细胞表面协同刺激分子的表达,减少炎性细胞及炎性介质的产生及分泌,减轻哮喘的变应性炎症,从而减少哮喘的发作的设想,对穴位敷贴疗法治疗哮喘可能与DC相关的免疫机制进行初步探讨。研究目的：验证“冬病夏治”穴位敷贴疗法能通过下调树突状细胞的表达,抑制DC细胞表面协同刺激分子的表达,减少炎性细胞及炎性介质的产生及分泌,减轻哮喘的变应性炎症,从而减少哮喘的发作的设想。从DC相关的免疫机制方面来阐释穴位敷贴治疗哮喘的理论依据。研究方法：用高浓度卵清蛋白(VOA)致敏大鼠,低浓度OVA激发建立过敏哮喘大鼠模型,用ELISA方法检测各组大鼠血清中OVA特异的IgE水平,比较穴位敷贴组,哮喘模型组,正常对照组之间有无差异；同用此法检测气管肺泡灌洗液(BALF)中IL-4、IL-5水平,比较各组之间有无差异。通过大鼠肺脏DC稳定的诱导与培养方法,用流式细胞仪检测肺脏DC表面分子CD11cCD80、CD11cCD86的表达,比较各组树状突细胞表面协同刺激分子表达水平有无差异。研究结果：与正常对照组相比,穴位敷贴组大鼠血清中VOA特异的IgE水平无明显差异(p0.05), BALF中IL-4、IL-5水平无明显差异(p0.05),树状突细胞表面协同刺激分子CD86/CD80表达水平无差异(p0.05)。与哮喘模型组相比,穴位敷贴组大鼠VOA特异的IgE水平有差异(p0.05),BALF中IL-4、IL-5水平有差异(p0.05),树状突细胞表面协同刺激分子CD86/CD80表达水平有差异(p(0.05)研究结论：穴位敷贴疗法能通过下调树突状细胞表面协同刺激分子的表达,减少炎症细胞及炎性介质的产生,缓解哮喘变应性炎症,从而减少哮喘的发作。
[Abstract]:Background: bronchial asthma is the most common chronic respiratory disease in children. It is caused by the interaction of various inflammatory cells and inflammatory mediators, which is caused by chronic inflammatory diseases of the respiratory tract in recent years. Some scholars have suggested that the main cause of recurrent bronchial asthma is the immune aberration of the body. And the most important immune abnormality is the disequilibrium between the number of Thl/Th2 cells and their function [1] .DC is the most potent of the antigen-presenting cells we've found so far. In addition to its unique ability to stimulate the proliferation of initial T cells, it can directly interfere with the differentiation direction of Th0 cells. This caused the imbalance of the number of Thl/Th2 cells and caused asthma attack [2] .Acupoint application therapy of "winter disease and summer treatment" is the most characteristic treatment method in the external treatment of traditional Chinese medicine, and it is one of the most typical representative prescriptions of external medicine preparations. Its mechanism is to achieve the goal of prevention and treatment of asthma through the dual effects of drugs on acupoints stimulation and permeation of effective components through skin [3]. This experiment combined with the previous related DC induction, culture and abnormal indexes in asthmatic rats. It is suggested that acupoint application therapy of "winter disease and summer treatment" can inhibit the expression of co-stimulating molecules on DC cells, reduce the production and secretion of inflammatory cells and inflammatory mediators, and alleviate allergic inflammation of asthma by down-regulating the expression of dendritic cells, inhibiting the expression of costimulatory molecules on the surface of DC cells, and reducing the secretion of inflammatory mediators. In order to reduce the attack of asthma, the immune mechanism related to DC in the treatment of asthma with acupoint application therapy was preliminarily discussed. Objective: to verify that acupoint application therapy of "winter disease and summer treatment" can down-regulate the expression of dendritic cells. Inhibiting the expression of costimulatory molecules on the surface of DC cells, reducing the production and secretion of inflammatory cells and inflammatory mediators, and alleviating allergic inflammation of asthma. In order to reduce the attack of asthma, the theoretical basis of acupoint application for asthma was explained from the immunological mechanism related to DC. Methods: sensitized rats were sensitized with high concentration of ovalbumin (ovalbumin) and induced by low concentration of OVA to establish an allergic rat model of asthma. The level of OVA specific IgE in serum of rats in each group was detected by ELISA method, and the level of IL-4 IL-5 in bronchoalveolar lavage fluid (BALF) was detected by this method, and the difference was found between acupoint application group, asthma model group and normal control group. The expression of CD11cCD80, CD11cCD86, a molecule on the surface of lung DC, was detected by flow cytometry through the method of stable induction and culture of rat lung DC, and the expression of CD11cCD80 and CD11cCD86 on the surface of pulmonary DC was detected by flow cytometry. To compare the expression level of co-stimulatory molecules on dendritic cells in each group. Results: compared with the normal control group, There was no significant difference in serum VOA specific IgE level in acupoint application group, no significant difference in IL-4 and IL-5 levels in BALF, and no difference in CD86/CD80 expression level on dendritic cells surface. Compared with asthmatic model group, there was no significant difference in the expression of CD86/CD80 between the two groups. There are differences in the level of IL-4 and IL-5 in VOA of rats in acupoint application group. Conclusion: acupoint application therapy can down-regulate dendritic cell surface through down-regulating dendritic cell surface. The expression of costimulatory molecules, Reduce the production of inflammatory cells and inflammatory mediators, alleviate allergic asthma inflammation, thereby reducing asthma attacks.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245

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