背景:连花清咳(LHQK)是一种治疗急性气管支气管炎的有效中药。在这项研究中,我们评估了LHQK治疗慢性阻塞性肺疾病急性加重期(AECOPD)气道黏液高分泌的有效性.
方法:雄性Wistar大鼠暴露香烟烟雾(CS)12周(80支/天,5天/周,持续12周)和气管内脂多糖(LPS)暴露(200μg,在第1、14和84天)。将大鼠分为六组:对照组(室内空气暴露),模型(CS+LPS暴露),LHQK(LHQK-L,LHQK-M,和LHQK-H),和阳性对照组(氨溴索)。H&E染色,和AB-PAS染色用于评估肺组织病理学,炎症反应,和杯状细胞增生。RT-qPCR,免疫组织化学,免疫荧光和ELISA用于分析转录,体内和体外人气道上皮细胞系NCI-H292中与粘液产生相关的蛋白质的表达和分泌。预测和筛选LHQK的活性成分,采用网络药理学分析和NF-κB报告系统分析。
结果:LHQK治疗可改善AECOPD引起的肺结构损伤,炎性细胞浸润,和促炎细胞因子的产生。AB-PAS和CCSP和Muc5ac抗体免疫荧光染色显示LHQK减少杯状细胞增生,可能通过抑制Club细胞转分化为杯状细胞。RT-qPCR和Muc5ac和APQ5的免疫组织化学显示,LHQK通过抑制体内和体外Muc5ac转录和高分泌来调节粘液稳态,并维持Muc5ac和AQP5表达之间的平衡。网络药理学分析和NF-κB荧光素酶报告系统分析提供了对LHQK活性成分的见解,这些成分可能有助于控制气道粘液高分泌和调节炎症。
结论:LHQK通过减轻炎症在AECOPD中表现出治疗作用,抑制杯状细胞增生,防止俱乐部细胞转分化,减少Muc5ac分泌过多,和调节气道粘液稳态。这些发现支持LHQK作为AECOPD潜在治疗的临床应用。
BACKGROUND: Lianhua Qingke (LHQK) is an effective traditional Chinese medicine used for treating acute tracheobronchitis. In this study, we evaluated the effectiveness of LHQK in managing airway mucus hypersecretion in the acute exacerbation of chronic obstructive pulmonary disease (AECOPD).
METHODS: The AECOPD model was established by subjecting male Wistar rats to 12 weeks of cigarette smoke (CS) exposure (80 cigarettes/day, 5 days/week for 12 weeks) and intratracheal lipopolysaccharide (LPS) exposure (200 μg, on days 1, 14, and 84). The rats were divided into six groups: control (room air exposure), model (CS + LPS exposure), LHQK (LHQK-L, LHQK-M, and LHQK-H), and a positive control group (Ambroxol). H&E staining, and AB-PAS staining were used to evaluate lung tissue pathology, inflammatory responses, and goblet cell hyperplasia. RT-qPCR, immunohistochemistry, immunofluorescence and ELISA were utilized to analyze the transcription, expression and secretion of proteins related to mucus production in vivo and in the human airway epithelial cell line NCI-H292 in vitro. To predict and screen the active ingredients of LHQK, network pharmacology analysis and NF-κB reporter system analysis were employed.
RESULTS: LHQK treatment could ameliorate AECOPD-triggered pulmonary structure damage, inflammatory cell infiltration, and pro-inflammatory cytokine production. AB-PAS and immunofluorescence staining with CCSP and Muc5ac antibodies showed that LHQK reduced goblet cell hyperplasia, probably by inhibiting the transdifferentiation of Club cells into goblet cells. RT-qPCR and immunohistochemistry of Muc5ac and APQ5 showed that LHQK modulated mucus homeostasis by suppressing Muc5ac transcription and hypersecretion in vivo and in vitro, and maintaining the balance between Muc5ac and AQP5 expression. Network pharmacology analysis and NF-κB luciferase reporter system analysis provided insights into the active ingredients of LHQK that may help control airway mucus hypersecretion and regulate inflammation.
CONCLUSIONS: LHQK demonstrated therapeutic effects in AECOPD by reducing inflammation, suppressing goblet cell hyperplasia, preventing Club cell transdifferentiation, reducing Muc5ac hypersecretion, and modulating airway mucus homeostasis. These findings support the clinical use of LHQK as a potential treatment for AECOPD.