基于“肝肺同治”理论的加味过敏煎调控哮喘小鼠Th细胞分化及其信号通路的研究

As one of the recognized medical challenges in the world, allergic asthma was listed as one of the four chronic and stubborn diseases by WHO. Modified Guomin Decoction (for anti-allergy) has been proven to be effective in allergy treatment, but its mechanism has yet to be disclosed. Therefore, in-depth studies and systematic development of Modified Guomin Decoction based on its effect of asthma relieving and anti-inflammation, may result in a new, safe and effective drug for asthma. However, recent studies of allergic asthma treatment with Modified Guomin Decoction mainly focus on simple, superficial evaluation of the effect of the drug on live animals, and studies of mechanism at organic, proteic, molecular and genetic levels are absent. On the basis of our previous results, we intend to, in this study, carry out in-depth exploration of regulation of Th cell differentiation and JAK-STAT signaling pathway in mice with asthma by Modified Guomin Decoction, using asthma mouse models induced by OVA as well as flow cytometry, protein chips, real time fluorescence quantitative PCR, WB, IHC and other modern molecular biology and immunological technologies, so as to elucidate the immunologically molecular mechanism and target of allergy and asthma relieving by Modified Guomin Decoction. Also, this study will be helpful in providing a scientific, reliable experimental basis for development of Modified Guomin Decoction into a new, safe and effective anti-asthma drug.

过敏性哮喘是世界公认的医学难题，被世界卫生组织列为疾病中四大顽症之一。加味过敏煎是防治过敏性疾病的有效方剂，但其机制尚未阐明。因此，结合加味过敏煎具有平喘、抗炎等作用，对加味过敏煎加以深入研究和系统开发很有可能得到新型、安全有效的治疗哮喘的新药。但从目前研究来看，对于加味过敏煎治疗过敏性哮喘的研究仅集中于对整体动物的简单浅层药效评价，缺乏从器官、蛋白、分子和基因水平的机制研究。因此，本研究拟在我们前期研究成果的基础上，采用OVA诱导的哮喘小鼠模型，进一步运用流式细胞术、蛋白芯片、实时荧光定量PCR、WB、IHC等现代分子生物学、免疫学技术对加味过敏煎调控哮喘小鼠Th细胞分化及对JAK-STAT信号通路的影响做深入探讨，以明确加味过敏煎抗敏平喘的免疫分子机制及作用靶点。为阐明加味过敏煎治疗哮喘作用提供科学可靠的实验依据，为把加味过敏煎开发为新型、安全、有效地拮抗哮喘的药物提供实验基础。

过敏性哮喘是目前的医学难题，给人类健康造成很大的影响，中医治疗过敏性哮喘有显著优势。本项目提出从“肝肺同治”角度治疗哮喘的新理论，将风邪引入过敏性疾病的病机当中，认为风邪是引发哮喘的先导，肝肺功能失常是引发哮喘的关键病机。结合现代医学与传统医学对哮喘的不同认识与理解，将中医的“肝肺同治”哮喘理论与现代医学关于哮喘发生的免疫学机制结合起来，对有临床实践基础的加味过敏煎进行系统的药效和药理学研究，在验证中医治疗学理论的同时，又可以开发中药复方新药，其成果可指导临床过敏性疾病（特别是过敏性哮喘）的预防与治疗，具有重要意义。.我们的研究结果表明：（1）药效方面：加味过敏煎通过多途径减少哮喘小鼠体内炎症细胞浸润，改变血液中细胞因子含量，减轻气道炎症，改善哮喘小鼠呼吸功能，发挥平喘、抗过敏的作用。（2）机制方面：加味过敏煎可能是通过抑制JAK-STAT信号通路中STAT6的活化及蛋白表达、增强STAT4的活化及蛋白表达，下调Th2、Th17型细胞因子及其相关转录因子的基因表达，上调Th1、Treg型细胞因子及其相关转录因子的基因表达，调节Th细胞向Th1、Th2、Th17和Treg细胞分化过程，扭转哮喘小鼠Th1/Th2细胞失衡，平衡Th17/Treg细胞。同时加味过敏煎还能下调哮喘小鼠肺组织凋亡抑制因子Bcl-2含量，上调促凋亡因子Bax含量；促进哮喘小鼠肺组织EOS凋亡，抑制EOS的浸润，减轻气道炎症反应。