Abstract

Ethnopharmacological relevance Linggan Wuwei Jiangxin Decoction (LWJD), a classical formula documented in Zhang Zhongjing's Jin Gui Yao Lue , composed of multiple herbal components, is widely used for respiratory diseases associated with cold-phlegm obstructing the lungs, such as Chronic Obstructive Pulmonary Disease (COPD) and bronchial asthma, demonstrating significant clinical efficacy. However, the mechanisms underlying its therapeutic effects on these conditions remain to be elucidated. Aim of the study This study aims to investigate the mechanisms by which LWJD alleviates airway inflammation in COPD-like model through network pharmacology analysis and experimental validation, providing a theoretical basis for its clinical application. Materials and methods Core targets of LWJD for COPD treatment were predicted using network pharmacology analysis and protein-protein interaction network prediction. Key signaling pathways of LWJD action were identified through KEGG pathway and GO enrichment analyses. A COPD model was established by cigarette smoke exposure and lipopolysaccharide (LPS) injection to simulate the disease. Inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF) and serum were measured by ELISA, while inflammatory cell proportions were evaluated via routine blood tests. Pathological changes in tracheal and lung tissues were examined using hematoxylin and eosin (H&E) and Masson staining. Expression of relevant mRNAs and proteins in lung tissue was assessed by quantitative real-time reverse transcription PCR (qRT-PCR) and Western blotting. To further investigate LWJD's anti-inflammatory mechanisms, LPS-induced inflammation was modeled in BEAS-2B human bronchial epithelial cells. The regulatory effects of LWJD on the AGE-RAGE signaling pathway were evaluated by qRT-PCR and Western blotting. Immunofluorescence detected nuclear translocation of downstream NF-κB p65. Reactive oxygen species (ROS) levels in cells were quantified by flow cytomet