基于NLRP3炎症小体介导的细胞焦亡探讨COPD相关性肺动脉高压大鼠血管重塑机制及“饮瘀同治”法干预作用

Pulmonary hypertension is an important complication of chronic obstructive pulmonary disease and vascular remodeling is an important sign of pulmonary hypertension. It is thought that immune inflammation plays an important role in the initiation and formation of COPD-associated pulmonary hypertension. Chronic inflammation in COPD may be associated with an adaptive immune response to congenital abnormality of immunity.Pyroptosis is a programmed cell inflammatory necrosis. NLRP3 inflammasome, as the most important inflammasomes which mediated pyroptosis, was closely related to the airway and vascular inflammation of COPD and PAH, and can affect the adaptive immunity. We speculate that vascular remodeling of COPD-PAH may be associated with an NLRP3 inflammasome-mediated pyroptosis and adaptive immunity caused by it .The ingredients of Chinese medicine with the function of activating blood and removing stasis can inhibit the NLRP3 inflammatory pathway and improve pulmonary vascular remodeling,and "Yin Yu Tong Zhi" for the treatment of pulmonary vascular remodeling through inhibition of NLRP3 inflammasome-mediated pyroptosis remains to be explored.In this study, COPD-PAH rats and alveolar macrophages were used to observe the effect of NLRP3 inflammasome-mediated pyroptosis of alveolar macrophages on T cell subsets and pulmonary vascular remodeling to reveal the molecular mechanism of vascular remodeling in COPD-PAH rats, and to explore the target of "Yin Yu Tong Zhi" improving the pulmonary vascular remodeling.

肺动脉高压是慢性阻塞性肺疾病重要并发症，肺血管重塑是肺动脉高压重要标志。目前认为免疫炎症在COPD相关性肺动脉高压起始和形成过程中起着重要的作用，COPD慢性炎症可能与先天性免疫异常导致适应性免疫反应有关。细胞焦亡是一种程序性细胞炎症坏死，NLRP3炎性小体作为介导细胞焦亡最重要的炎症小体与COPD及PAH气道和血管炎症密切相关，并能影响适应性免疫。我们推测COPD-PAH血管重塑可能与NLRP3炎症小体介导的细胞焦亡及其引起的适应性免疫有关。活血化瘀中药能抑制NLRP3炎症小体通路和改善肺血管重塑，“饮瘀同治”法是否通过抑制NLRP3炎症小体介导的细胞焦亡改善肺血管重塑有待探索。拟以COPD-PAH大鼠及肺泡巨噬细胞为研究对象，观察NLRP3炎症小体介导的细胞焦亡对T淋巴细胞亚群及肺血管重塑的影响，揭示COPD-PAH大鼠血管重塑可能机制及“饮瘀同治”法改善肺血管重塑作用机理。

慢阻肺相关性肺动脉高压是属于肺疾病相关性肺动脉高压，是慢阻肺进展的常见并发症。为探讨经典细胞焦亡途径与非经典途径介导的细胞焦亡与肺动脉高压的关系以及复方葶苈子汤对其干预机制，我们体内采用香烟烟雾联合气道滴注脂多糖建立大鼠COPD-PAH模型，复方葶苈子汤以方测证验证模型属性，从细胞焦亡诱发的炎症级联反应探讨其与COPD-PAH的关系及复方葶苈子汤对其的干预机理。体外实验采用香烟烟雾联合脂多糖诱导NR8383巨噬细胞、PMVEC（大鼠肺血管内皮细胞）细胞焦亡，复方葶苈子汤含药血清进行干预。研究结果证实复方葶苈子汤可以改善COPD-PAH大鼠肺功能、肺动脉压力、炎症反应以及气管、肺泡及肺动脉的病变情况。香烟烟雾联合脂多糖诱导细胞焦亡发生，体内体外实验模型均存在细胞细胞焦亡相关指标（NLRP3、caspase1、caspase11、GSDMD、IL-1β、IL-10、IL-18）等高表达，在复方葶苈子汤干预下表达降低，提示复方葶苈子汤可通过抑制NLRP3炎症小体介导的细胞焦亡改善肺血管重塑。但是，在含药血清部分体外实验，未对中药进行质谱分析及直接使用中药过滤物干预细胞，存在一定不足，在接下来实验中我们课题组会进行进一步改正与完善。