补肺颗粒通过调控纤毛自噬降低COPD细菌负荷预防急性加重的“扶正”机制

COPD patients with long-term recurrent cough and sputum may increase the risk of acute exacerbation (AECOPD) and death. Ciliary dysfunction, hypersecretion of mucus and nausea cycle of bacterial load is the main reason. The project team proposed the existence of healthy qi deficiency and pathogens strong in stable COPD, and its treatment needs integrated regulation / Precision promotion. And Bufei granule can improve symptoms and reduce exacerbations, which related to the integrated regulation of airway mucus hypersecretion and reduction of bacterial load, but it can not clarify its mechanism of Precision promotion. Cilia autophagy is associated with impaired cilia autophagy clearance. The clarification of cell autophagy mechanism provides a new idea for revealing positive qi deficiency and phlegm of TCM. The project will establish AECOPD model by making rats cigarette smoked and injecting Streptococcus pneumoniae in rats tracheal, and discuss the mechanism that how the Chinese medicinal herbs for strengthening healthy Qi, herbs for strengthening healthy Qi and eliminating pathogens, and Western medicine for expectorant / antioxidant strengthen healthy qi of rats with bacterial infection in stable COPD based on cilia autophagy. Airway epithelial cells of Normal human and COPD for the study, the team will observe the effect and mechanism of Bufei granule on the cilia autophagy of lung airway epithelial cells by using gas-liquid culture, CCK8, MDC, quantitative real-time PCR, et al. and explore its internal mechanism. The proiect research "strengthening healthy qi" mechanism of Bufei granule in overall and cellelar level, and expound the nature and content of overall regulatory / Precision promotion scientifically, Which will enrich TCM theory of Healthy Qi and pathogens, and innovate COPD pathogenesis and treatment.

慢阻肺患者长期咳嗽咳痰可增加急性加重（AECOPD）和死亡风险，纤毛功能障碍、粘液高分泌和细菌负荷的恶性循环是主要原因。项目组提出COPD稳定期存在本虚标实，治疗需整合调控、精准扶正，补肺颗粒可改善症状且减少急性加重，与整合调控粘液高分泌、细菌负荷及炎症有关，但无法阐明其精准扶正机制。纤毛自噬与气道防御功能受损相关，细胞自噬为揭示中医气虚痰瘀提供思路。项目将通过在香烟烟熏大鼠气管内注入肺炎链球菌建立AECOPD模型，基于纤毛自噬探讨中药单纯扶正和扶正祛邪及西药祛痰/抗氧化对细菌感染后COPD稳定期动物模型的“扶正”机制。以正常人体和COPD患者气道上皮细胞为研究对象，采用气-液面共培养、CCK8、MDC和实时定量PCR等方法，观察补肺颗粒对气道上皮细胞自噬的影响和机制。从整体和细胞水平研究补肺颗粒的“扶正”机制，促进整合调控、精准扶正作用的科学表达，丰富中医正邪理论，创新COPD病机治法。

慢性阻塞性肺部疾病（COPD）是一种严重危害人类健康的常见病、多发病，COPD急性加重是慢阻肺患者死亡的重要因素。细胞自噬在COPD的发病中起着重要且复杂的作用，气道上皮细胞自噬与其功能受损相关，通过对气道上皮细胞炎症因子、蛋白和自噬途径的调控，将有利于改善气道防御功能，并成为 COPD 潜在的新的治疗靶点。. 本研究从体内、体外两方面分别基于细胞自噬探索了补肺颗粒预防COPD急性加重的“扶正”作用机制，结果证实补肺颗粒治疗COPD的扶正机制是抑制肺部炎症因子及自噬相关蛋白的表达，进而抑制气道上皮的自噬，保护气道功能，提高机体的防御能力，达到“扶正”的效果。研究一开展动物实验，建立大鼠COPD模型，通过给予扶正中药单用、补肺颗粒、西药抗氧化三种干预措施，从气道功能、信号通路失衡、病原学等方面，阐述其内在机制。研究二以人气道上皮细胞为研究对象，分别设立空白组、空白+补肺颗粒组、模型组、补肺颗粒干预组，干预24h之后，检测细胞上清液中的细胞因子或蛋白的表达情况，以阐述补肺颗粒对气道上皮细胞自噬调控的效果与机制。结果发现扶正中药单用及补肺颗粒合方均能降低COPD下气道细菌负荷，抑制肺部炎症因子（TLR4 、IL-1β、MUC5AC 、HDAC6、Caspase-3 ）的过度表达，进而抑制其诱发的一系列细胞自噬，减少对气道上皮的损伤，以延缓肺功能的下降，提高机体的防御能力，减少急性加重发生的次数，对机体起保护作用。基于网络药理学的研究，初步明确了补肺颗粒中含有多种有效成分，其通过抑制支气管平滑肌收缩、激活环磷酸腺苷通路、缓解氧化应激来发挥作用。