止嗽散调控TRPV1和TRPA1信号通路治疗咳嗽变异性哮喘机制研究

Cough variant asthma (CVA) is a chronic cough as the main performance of the special type of asthma, some patients will develop into a typical asthma. TRPV1 and TRPA1 was proved to be cough reflex "switch", played an important role in asthma and cough, but for the mechanism of action of CVA is unclear. We put forward "micro poverty cough" is the core of the CVA pathogenesis science hypothesis, according to the pathogenesis, we use the Zhisou powder treatment in the previous study achievement obtained good curative effect, and reduce airway responsiveness and inflammatory factor, may Zhisou powder are scattered in the TRPV1 and TRPA1 was signaling pathways regulating works. To prove this hypothesis, this research intends to copy the CVA model of guinea pigs, from coughing reaction, airway reactivity, histopathological and molecular biology, the study of the pathogenesis of CVA, from TRPV1 and TRPA1 was signaling pathways of gene expression level, protein molecules analysis is sensitive to cold channel TRPV1, thermal sensitive TRPA1 was signaling pathways and CVA relationship; Application Zhisou powder away intervention CVA guinea pigs at the same time, to explore traditional Chinese medicine (TCM) on inflammatory factor and TRPV1 and TRPA1 was the influence of the signal path, reveal Zhisou powder treatment of CVA mechanism and link.

咳嗽变异性哮喘（CVA）是以慢性咳嗽为主要表现的特殊哮喘类型，部分病人将发展成典型哮喘。TRPV1和TRPA1已被证明是咳嗽反射的“开关”，在哮喘和咳嗽中扮演了重要角色，但对CVA的作用机制尚不清楚。我们提出“微寒微咳”是CVA的核心病机假说，针对这一病机，我们在既往研究中应用止嗽散治疗CVA取得良好疗效，实验研究证实润肺法可降低气道反应性 和炎症因子，但TRPV1和TRPA1信号通路调控机制及止嗽散干预机制不明。为证实这一假说，本 课题组拟复制CVA豚鼠模型，从咳嗽反应、气道反应性、组织病理学及分子生物学方面，研究CVA的发病机制，从TRPV1及TRPA1信号通路的基因蛋白分子表达水平，分析寒敏感通道TRPV1、热敏感通道TRPA1信号通路与CVA的关系；同时探讨止嗽散干预CVA豚鼠，探索中药对炎症因子及TRPV1及TRPA1信号通路的影响，揭示止嗽散干预CVA的作用机理和环节。

多种炎症细胞、炎症介质共同参与的气道慢性炎症是咳嗽变异性哮喘（CVA）的重要病理基础；而TRPV1、TRPA1对诸多炎症细胞及炎症介质起重要调控作用，可能对CVA气道慢性炎症的发生发展有重要意义。CVA患者常表现为对冷空气敏感，这与中医“寒邪”相关，因此，本课题组提出“微寒微咳”是CVA重要病机的假说。. 本研究以TRPV1、TRPA1在CVA中的作用为切入点，观察TRPV1、TRPA1在气道慢性炎症形成中的作用，探讨TRPV1和TRPA1信号通路在CVA发病机制中的作用；基于“微寒微咳”病机假说，观察以温润止咳立法的止嗽散对CVA豚鼠和大鼠气道反应性、支气管肺组织病理形态学、血清及支气管肺泡灌洗液（BALF）炎症细胞/介质、肺组织TRPV1、TRPA1、TGF-β1表达的影响，以揭示其治疗CVA的机理，阐明“微寒微咳”的病机假说。. 研究结果显示：（1）豚鼠实验研究：与空白对照组比较，模型组豚鼠肺组织病理损伤、炎症细胞浸润明显，BALF EOS%明显升高，肺组织TRPV1、TRPA1蛋白基因表达有增加趋势；与模型组比较，止嗽散、糖皮质激素、TRPV1拮抗剂、TRPA1拮抗剂、TRPV1+TRPA1拮抗剂组豚鼠肺组织病理损伤、炎症细胞浸润减轻，BALF EOS%、肺组织TRPV1、TRPA1蛋白基因表达呈减少趋势。（2）大鼠实验研究：与空白对照组比较，模型组大鼠气道阻力明显升高，支气管及肺组织病理损伤、炎症细胞浸润明显，血清及BALF ECP、IL-4、IL-5、TGF-β1水平、肺组织TGF-β1蛋白表达明显升高，肺组织TRPV1、TRPA1蛋白表达有增加趋势；与模型组比较，止嗽散、激素、各拮抗剂组大鼠气道阻力、血清及BALF ECP、IL-4、IL-5、TGF-β1降低或有降低趋势，支气管肺组织病理损伤、炎症细胞浸润减轻，肺组织TRPV1、TRPA1、TGF-β1蛋白表达呈减少趋势。. 本研究初步证实了TRPV1、TRPA1的异常激活可能与CVA发病相关，拮抗TRPV1、TRPA1可能对CVA有治疗作用，以及止嗽散可能通过调控CVA豚鼠、大鼠肺组织中TRPV1、TRPA1来控制气道慢性炎症，进而有效治疗CVA的作用机制，阐明了“微寒微咳”在CVA中的科学内涵，并为止嗽散的临床应用提供了实验依据。