Analyzing mechanisms of qing fei bao yuan decoction granules in treating COPD based on LC-MS, network pharmacology and in vivo methods

Abstract

Background and aim: The current therapeutic interventions of chronic obstructive pulmonary disease offer only partial alleviation of symptoms, leaving the majority of patients with persistent and significant clinical manifestations. This investigation seeks to elucidate the underlying pharmacological mechanisms of Qing Fei Bao Yuan Decoction (QFBYD) employing a multidisciplinary approach that includes network pharmacology and molecular docking techniques.

Experimental procedure: The QFBYD formulation were subjected to mass spectrometry analysis, while critical compounds and biological targets were subsequently identified through the TCMSP database. Disease- and drug-specific targets were collated from a plethora of databases, including Batman-TCM, Stitch, Swiss Target Prediction and GeneCards. GO and KEGG pathways were analyzed for the collected targets. A PPI network was constructed using STRING database to isolate core targets. Molecular docking was executed using Auto Dock Tools and PyMOL software, and an animal model of COPD was developed for experimental validation.

Results and conclusions: Seven salient compounds and five core biological targets were ascertained through our analysis. Additionally, four compounds demonstrated high-affinity binding to the identified targets. Pathways involving bacterial endotoxin response, oxidative stress regulation, and endothelial cell migration were significantly enriched according to the KEGG database. Animal models substantiated that QFBYD ameliorated pathological hallmarks, enhanced respiratory functionality, mitigated overexpression of pro-inflammatory cytokines, augmented the antioxidant defense mechanism, and suppressed the hyperactivity of the five core targets. The efficacy of QFBYD in COPD treatment may be primarily attributed to its role in moderating inflammatory responses and rectifying oxidative imbalances.

Keywords: Chronic obstructive pulmonary emphysema; Experimental analysis; Mechanism; Molecular docking; network pharmacology.

Graphical abstract

Fig. 1

Total ion chromatograms (TIC) from QFBYD. (A) TIC (+ESI TCC) of QFBYD; (B)TIC (-ESI TCC) of QFBYD.

Fig. 2

Screening of potential targets and the construction of the "disease-component-potential target" network. (A) A Venn diagram illustrating the intersectionality among Disease-Related Targets and Compounds is presented. In both the GeneCards and DisGeNet databases, 519 common genes associated with COPD were initially isolated, and subsequent intersection with compound-specific target genes resulted in the identification of 83 potential targets. (B) A network map elucidating the complex relationships among Disease-Related Targets and Compounds was generated. Post-analysis, these 83 potential targets demonstrated a significant relationship with the aforementioned seven critical compounds, culminating in the construction of a comprehensive "disease-ingredient-potential target" network via Cytoscape v3.9.1.

Fig. 3

Screening of the protein interaction network and core targets of QFBYD acting on COPD. The aforementioned 83 putative targets were uploaded into the STRING database for the acquisition of a comprehensive PPI network diagram, which was subsequently imported into Cytoscape v3.9.1 for advanced topological scrutiny. Core targets within the PPI network were selected based on analyses conducted via the Maximal Clique Centrality (MCC) algorithm, facilitated by the CytoHubba plug-in. The apex targets were identified as TNF, AKT1, HIF1α, IL1β, and PTGS2.

Fig. 4

(A) GO terms of top 10 functional terms were selected. (B) KEGG pathway gathering of top 20 pathway were selected. The more obvious the red, the more important the pathway.

Fig. 5

Molecular docking of the core target to the compounds. (A) TNF and Liquiritin; (B) HIF1ɑ and Liquiritin; (C)IL-1β and Liquiritin; (D) PTGS2 and Liquiritin; (E) AKT1 and Hesperetin; (F) HIF1ɑ and Hesperetin; (G) PTGS2 and α-Linolenic acid; (H)PTGS2 and Oxyresveratrol.

Fig. 6

Lung pathological findings in COPD rats. (A) Selection of three random specimens from the model group at 4, 8, and 12-week intervals, with histopathological alterations observed via Hematoxylin and Eosin (HE) staining; (B) High magnification (100 × ) of lung tissue post-QFBYD treatment. (C–H) Respiratory parameters were measured in awake, freely mobile animals. QFBYD elicited significant amelioration in lung function among COPD afflicted rats (^##P < 0.01 vs. Ctrl group; *P < 0.05 and **P < 0.01 vs. Model group, n = 10).

Fig. 7

(A–F) ELISA was used to monitor the IL-1β, IL-6, TNF-ɑ, IL-17, IL-8 and Ferritin plasma levels in COPD rats (^##P < 0.01 vs. Ctrl group; *P < 0.05 and **P < 0.01 vs. Model group, n = 10).

Fig. 8

(A–D) ELISA was used to get the SOD, MDA, Catalase and GPX plasma levels in COPD rats. (E–F) AKT, HIF1ɑ and PTGS2 levels in COPD rats (^##P < 0.01 vs. Ctrl group; *P < 0.05 and **P < 0.01 vs. Model group, n = 10).