Network Pharmacology and Experimental Verification Reveal the Regulatory Mechanism of Chuanbeimu in Treating Chronic Obstructive Pulmonary Disease

Abstract

Background: Chronic obstructive pulmonary disease (COPD) is a common respiratory disorder in pulmonology. Chuanbeimu (CBM) is a traditional Chinese medicinal herb for treating COPD and has been widely utilized in clinical practice. However, the mechanism of CBM in the treatment of COPD remains incompletely understood. This study aims to investigate the underlying therapeutic mechanism of CBM for COPD using network pharmacology and experimental approaches.

Methods: Active ingredients and their targets were obtained from the Traditional Chinese Medicine Systems Pharmacology database. COPD-associated targets were retrieved from the GeneCards database. The common targets for CBM and COPD were identified through Venn diagram analysis. Protein-protein interaction (PPI) networks and disease-herb-ingredient-target networks were constructed. Subsequently, the results of the network pharmacology were validated by molecular docking and in vitro experiments.

Results: Seven active ingredients and 32 potential targets for CBM were identified as closely associated with COPD. The results of the disease-herb-ingredient-target network and PPI network showed that peimisine emerged as the core ingredient, and SRC, ADRB2, MMP2, and NOS3 were the potential targets for CBM in treating COPD. Molecular docking analysis confirmed that peimisine exhibited high binding affinity with SRC, ADRB2, MMP2, and NOS3. In vitro experiments demonstrated that peimisine significantly upregulated the expression of ADRB2 and NOS3 and downregulated the expression of SRC and MMP2.

Conclusion: These findings indicate that CBM may modulate the expression of SRC, ADRB2, MMP2, and NOS3, thereby exerting a protective effect against COPD.

Keywords: chronic obstructive pulmonary disease; chuanbeimu; experiment verification; molecular docking; network pharmacology.

Figure 1

The overall flow chart of this work.

Figure 2

Screening the targets of CBM and COPD. (A) Venn diagram showing the common targets of CBM in the treatment of COPD; (B) PPI network of the common targets; (C) Barplot of the common targets.

Figure 3

Functional enrichment analysis of the common targets. (A) Bubble chart of the biological process category terms; (B) Bubble chart of the molecular function category terms; (C) Bubble chart of the cellular component category terms; (D) Bubble chart showing the top 10 enriched KEGG terms.

Figure 4

Network construction of the herb, ingredient, and targets. (A) Network of the disease-herb-ingredient-target for the treatment of COPD using the CBM; (B) Peimisine and disease-target network; (C) Barplot of the 16 targets of peimisine.

Figure 5

Molecular docking analysis between peimisine and the targets. (A) SRC, (B) ADRB2, (C) MMP2, and (D) NOS3.

Figure 6

Establishment and evaluation of COPD cell model. (A) Cell proliferation was detected by CCK8 assay at 24 h and 48 h; (B and C) Detection of inflammatory factors; (D) Detection of cell apoptosis. All in vitro experiments were repeated three times independently. *p < 0.05 and ***p < 0.001.

Figure 7

To clarify the effect of Peimisine on target gene expression in COPD. Effect of Peimisine on cell viability at 24h (A). RT-qPCR was used to detect the mRNA expression levels of SRC (B), ADRB2 (C), MMP2 (D), and NOS3 (E). All experiments were repeated three times independently. *p < 0.05 and and ***p < 0.001.