益气温阳法调控变应性鼻炎NF-kB信号转导通路的研究

Allergic rhinitis (AR) is the common illness which is the serious harmful to people's health and the quality of life. Traditional Chinese Medicine(TCM) treatment has the unique advantage in curative effect. Tonifying qi and yang method in TCM is the fundamental therapy. The pathogenesis of AR is more complicacy, disproportionate of Th1/Th2, especially the High-expression of Th2 is the pathogenic key point of AR. IL-4、IL-5 are relative cytokine with Th2, which are controlled by NF-κB. That is indicated the signal transmission of NF-κB is important in regulation inflammation of AR.So we hypothesize that the mechanism of the therapy of tonifying qi and yang to cure AR is to inhibit the activity of NF-κB, the expression of IL-4、IL-5, and regulate the signal transmission of NF-κB. The subject of this study is the patients with AR of lung and kidney deficiency pattern. The test methods of ELISA, electrophoretic mobility shift assay, Western blot and RT-PCR are used to test those indexs: AR-related inflammatory cytokine, the expression level of upstream and downstream protein in signal transmission of NF-κB and the binding activity of DNA with NF-κB. And expound the mechanism how the above therapy treats control the signal transmission of NF-κB in AR. For providing the theoretical basis for the clinical treatment of AR. In order to further promote the use of tonifying qi and yang method in clinical treatment of AR.

AR是严重危害人们健康与生活质量的常见多发病。中医药治疗具有独特疗效优势，益气温阳法是一种基本治法。AR发病机制较为复杂，Th1/Th2细胞因子网络的失衡，尤其是Th2型细胞因子的异常高表达是发病的关键环节，IL-4、IL-5是与Th2相关最重要的细胞因子，而其表达受核因子-kB的调控，表明NF-kB是调控AR炎症的重要信号通路。据此提出假说：益气温阳法治疗AR的一个重要机制是抑制NF-kB的结合活性，降低IL-4、IL-5表达，调控NF-kB信号转导通路。本课题以AR肺肾气虚证患者为对象，运用ELISA法、凝胶迁移实验、蛋白印迹、RT-PCR技术等方法，检测AR相关炎性细胞因子、NF-kB信号通路的上、下游蛋白表达水平及与DNA的结合活性等指标，阐明益气温阳法调控AR NF-kB信号转导通路的作用机制，为其临床治疗AR提供理论依据。以此进一步推动益气温阳法在临床治疗AR的广泛应用。

本课题基本按计划顺利完成，并取得预期结果。课题以肺肾气虚型AR患者为研究对象，采用随机双盲安慰剂对照设计，通过ELISA检测、免疫组化、RT-PCR、Western Blot等技术，研究了益气温阳治法对肺肾气虚型AR患者Th1/Th2细胞因子IL-4、IL-5、IFN-的影响，以及对Th2上游调节信号蛋白NF-κB p65、NF-κB p-p65、IκBα、p-IκBα表达的影响。结果：试验组治疗后血清IL-5水平较试验组治疗前低（p＜0.05），而安慰剂组治疗前后无明显变化（p＞0.05）。试验组治疗后鼻腔粘膜NF-κB p-p65水平较试验组治疗前降低（p＜0.05），而安慰剂组治疗前后无明显变化（p＞0.05）。结论：益气温阳法治疗能降低肺肾气虚型AR患者血清IL-5水平，减轻鼻腔变态反应性炎症。益气温阳法治疗能降低鼻腔粘膜NF-κB p65蛋白的磷酸化水平，抑制NF-κB入核，从而抑制Th2细胞因子的转录，纠正AR患者Th1/Th2细胞因子网络的失衡状态。