基于TACE-EGFR信号通路探讨扶正祛邪法抑制COPD大鼠气道黏液高分泌的效应机制

Explore the effect and mechanism of Fuzheng Quxie method in inhibiting airway mucus hypersecretion in COPD rats based on the TACE-EGFR signaling pathway

关键词:  慢性阻塞性肺疾病  气道黏液高分泌  补肺颗粒  TACE-EGFR信号通路

Key words:  chronic obstructive pulmonary disease  airway mucus hypersecretion  Bufei Granules  TACE-EGFR signaling pathway

摘要:

目的 构建慢性阻塞性肺部疾病（COPD）稳定期大鼠模型，探讨扶正祛邪中药补肺颗粒干预肿瘤坏死因子转换酶-表皮生长因子受体（TACE-EGFR）信号通路，抑制气道黏液高分泌，降低气道细菌负荷的效应机制。方法 40只Wistar大鼠随机分为空白对照组、模型组、乙酰半胱氨酸组、扶正中药组、扶正祛邪中药组，除空白对照组外均采用香烟烟雾暴露结合气管内缓慢滴注肺炎链球菌琼脂悬液构建AECOPD大鼠模型，进入稳定期后进行1个月的药物干预，运用HE染色进行肺组织病理学分析，肺组织细菌菌落计数观察细菌负荷水平，ELISA法检测大鼠血清及肺泡灌洗液中TACE、转化生长因子-α（TGF-α）、黏蛋白5AC（MUC5AC）的含量，RT-PCR法、Western blot法检测TACE、EGFR mRNA及蛋白表达情况。结果 与空白对照组相比，模型组大鼠支气管结构出现明显损伤，肺组织匀浆稀释后接种于平板可见菌落密集满平板生长，大鼠血清及肺泡灌洗液TGF-α、TACE与MUC5AC水平显著升高（P < 0.01），大鼠肺组织EGFR、TACE mRNA及蛋白表达显著增多（P < 0.01）。与模型组相比，与扶正祛邪中药组大鼠支气管结构损伤及黏膜下炎细胞浸润减轻，接种后菌落数较模型组明显减少，大鼠血清及肺泡灌洗液TGF-α、TACE与MUC5AC水平显著降低（P < 0.05），大鼠肺组织EGFR、TACE mRNA及蛋白表达显著降低（P < 0.05）。结论 扶正祛邪中药补肺颗粒可能通过干预TACE-EGFR信号通路，抑制气道黏液高分泌，降低细菌负荷，综合发挥扶正祛邪效应。

Abstract:

Objective To establish a stable-phase chronic obstructive pulmonary disease (COPD) rat model and investigate the mechanism of Bufei Granules (BFKL), a Chinese herbal medicine for reinforcing healthy qi and eliminating pathogens, in inhibiting airway mucus hypersecretion and reducing bacterial load via the TACE-EGFR signaling pathway. Methods The 40 Wistar rats were randomly divided into five groups: blank control, model, acetylcysteine, reinforcing healthy qi herbal group (FZ), and reinforcing healthy qi-eliminating pathogens herbal group (FZQX). Except for the blank control group, all rats were subjected to cigarette smoke exposure combined with slow intratracheal instillation of Streptococcus pneumoniae agar suspension to establish an acute exacerbation of COPD (AECOPD) model. After stabilization, a 1-month drug intervention was administered. Pathological changes in lung tissue were analyzed via HE staining, bacterial load was assessed via colony counting, serum and bronchoalveolar lavage fluid (BALF) levels of TACE, TGF-α, and MUC5AC were measured using ELISA, and TACE/EGFR mRNA and protein expression were detected via RT-PCR and Western blot. Results Compared with the blank control group, the model group exhibited significant bronchial structural damage, dense bacterial colony growth in lung homogenate cultures, elevated serum and BALF levels of TGF-α, TACE, and MUC5AC (P < 0.01), and increased TACE/EGFR mRNA and protein expression in lung tissue (P < 0.01). Compared with the model group, the FZQX herbal group showed alleviated bronchial damage, reduced submucosal inflammatory cell infiltration, fewer bacterial colonies, decreased TGF-α, TACE, and MUC5AC levels (P < 0.05), and downregulated TACE/EGFR mRNA and protein expression (P < 0.05). Conclusion Bufei Granules may inhibit airway mucus hypersecretion and reduce bacterial load by modulating the TACE-EGFR signaling pathway, demonstrating comprehensive therapeutic effects by reinforcing healthy qi and eliminating pathogens.