探讨JAK/STAT通路调控慢性阻塞性肺疾病急性加重期（痰热阻肺）大鼠气道炎症及清金化痰颗粒干预实验研究

In AECOPD airway inflammation, are still lack of exact effective drug treatment,So further，the important of explore new drug is increasingly.In COPD patients with acute aggravating period is often shown as "phlegm" and "hot" is given priority to, and clip "stasis", "the real" Chinese medicine syndrome, "clearing heat and transforming phlegm, promoting blood circulation to remove blood stasis method" as the doctor of traditional Chinese medicine treatment of COPD acute aggravating period airway inflammation of the breakthrough point.This project use smoke edify poison + lipopolysaccharide airway shots is to establish an animal model of acute airway inflammation to simulate acute aggravating period COPD airway inflammatory change, choose clear gold phlegm soup as intervention measures to internationally recognized leucocyte total percentage of neutrophils and macrophages number, IL - β 1, IL - 6, IL - 8, TNF alpha, leukotriene B4, leukotriene C4 level, bronchial lung tissue of rats HE dyeing pathological changes as airway inflammation evaluation index, the protein chip technology, dynamic observation group rats serum and lung tissue JAK - STAT signaling pathways in immune network proteomics characteristics. Discuss AECOPD pathogenesis and clear phlegm gold particles intervention effect.

针对AECOPD气道炎症，至今尚缺乏确切有效的治疗药物，因此深入探索新药物显得日趋重要。在COPD 急性加重期患者往往表现为以"痰"、"热"为主兼夹“瘀”、“腑实”的中医证候，“清热化痰、活血化瘀法”作为中医治疗COPD 急性加重期气道炎症的切入点。本项目采用烟熏染毒+脂多糖气道注射法建立急性气道炎症动物模型以模拟COPD 急性加重期气道炎症改变，选用清金化痰汤作为干预措施，以国际公认的白细胞总数、中性粒细胞百分比、巨噬细胞数、IL-1β、IL-6、IL-8、TNF-α 、白三烯B4、白三烯C4水平、大鼠支气管肺组织HE 染色病理改变作为气道炎症的评价指标，采用蛋白质芯片技术，动态观测各组大鼠血清、肺组织中JAK-STAT信号途径免疫网络的蛋白质组学特征。探讨AECOPD发病机制及清金化痰颗粒的干预作用。

本项目采用ELISA、免疫组化、RT-PCR、western-blot及PCR芯片检测等现代分子生物学技术，以AECOPD（痰热郁肺）型大鼠为研究对象，以清金化痰颗粒干预各组模型大鼠后，观察各组大鼠肺组织病理病化、BALF液中炎症介质的变化、肺组织中JAK/STAT相关分子通路的的表达及大鼠肺组织中基因表达谱等方面开展相关研究，结果表明：（1）清金化痰颗粒可以使大鼠肺组织支气管壁变薄、充血水肿减轻,炎症细胞侵润减少；（2）大鼠BALF液中相关炎症介质水平（如IL-6、IL-8、IL-1β、TNF-α、LTB4及LTC4）均明显下降，清金化痰颗粒（尤其是高剂量组）对这种情况有一定的改善，其作用效果与罗红霉素相近；（3）大鼠肺组织中JAK/STAT相关分子通路STAT1、STAT3、JAK2、p-JAK2、p-STAT1、p-STAT3水平均有不同程度下调，而SCOS3水平上调,其分子作用机制可能与STAT1、STAT3、JAK2、p-JAK2、p-STAT1、p-STAT3的下调和SCOS3上调有关，降低大鼠肺组织中相关炎症介质的释放，最终达到减轻炎症反应的作用；（4）PCR芯片检测表明清金化痰颗粒可能是通过激活或抑制JAK/STAT 信号通路的相关基因来减轻炎性反应，后续研究将对相关上调/下调基因进行生物信息学分析并进行相关验证，进一步揭示其作用机制。