Exploring the Mechanism of Guben Pingchuan Granule in the Treatment of Bronchial Asthma

Summary

This protocol investigates the mechanism of action of Guben Pingchuan granules in bronchial asthma using network pharmacology, molecular docking, and in vivo. validation.

Abstract

This study investigates the mechanism of action of Guben Pingchuan granules in the treatment of bronchial asthma using network pharmacology, molecular docking, and experimental validation. Active ingredients and corresponding targets were identified using TCMSP, HERB, and SymMap databases. Bronchial asthma-related targets were collected from GeneCards, OMIM, TTD, DrugBank, and PharmGKB. Overlapping targets were identified, and component–target and protein–protein interaction (PPI) networks were constructed. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed with false discovery rate (FDR) correction. Molecular docking was conducted to evaluate binding affinity, followed by in vivo validation. A total of 121 active components and 234 drug-related targets were identified, yielding 155 overlapping targets. Core targets included TP53, AKT1, TNF, and IL6. KEGG analysis demonstrated significant enrichment of the PI3K–Akt signaling pathway, which was selected for experimental validation. Molecular docking revealed a strong binding affinity between core components and AKT1. In ovalbumin-induced asthmatic rats, Guben Pingchuan granule reduced serum IL-4 and IL-10 levels, increased FoxP3 expression in lung tissue, and upregulated PI3K and AKT mRNA levels. These findings indicate that Guben Pingchuan granule alleviates bronchial asthma by modulating the PI3K–Akt. signaling pathway, regulating inflammatory cytokines, and enhancing regulatory T cell function.

Introduction

Bronchial asthma is a heterogeneous chronic respiratory disease characterized by airway inflammation, airway hyperresponsiveness, mucus hypersecretion, and airway remodeling¹. Its pathogenesis involves complex interactions among immune cell activation, inflammatory mediators, and multiple signaling pathways. Clinically, asthma presents with recurrent wheezing, dyspnea, chest tightness, and cough. Airway inflammation is a central mechanism underlying disease onset and progression, characterized by increased secretion of pro-inflammatory cytokines such as IL-6, IL-4, IL-17, and TNF-α., infiltration of inflammatory cells inc....

Protocol

All animal procedures were approved by the Animal Ethics Committee of Liaoning University of Traditional Chinese Medicine (Approval No.: 21000042023058) and performed in accordance with institutional guidelines. The research tools used in this protocol are listed in the Table of Materials.

1. Databases and software

1. Access the following databases and software: TCMSP, HERB, SymMap, PubChem, STRING, GeneCards, OMIM, TTD, DrugBank, PharmGKB, DAVID, UniProt, and Cytoscape (version 3.9.1).

2. Acquisition of component targets

1. Retrieve chem....

Results

Screening of active ingredients
A total of 121 active components were identified from the TCMSP database after filtering for target activity. These included 23 components from Ephedra Herb, 7 from Psoralea, 19 from Apricot seed, 8 from Pseudostellaria root, 16 from Perilla fruit, 18 from Loquat leaf, 7 from Lilium, and 23 from Epimedium. (Table 2). Sixteen common components were identified, including luteolin, quercetin, β-sitosterol, kaem.......

Discussion

Bronchial asthma is a common disease in the respiratory department¹⁶. It is an obstructive airway disorder caused by allergens and airway hyperresponsiveness¹⁷. It has a high clinical incidence, is prone to recurrence, and is difficult to cure, significantly affecting patients' health and quality of life. The 2025 Global Initiative for Asthma (GINA) reports that inhaled glucocorticoid therapy is effective for asthma¹⁸; however, long-term use may .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
qPCR Kit	Novogene	NQ-001, 2×1mL	Detection of relative mRNA expression of target genes
aluminum hydroxide Al(OH)3	Shanghai Yubo Biotechnology Co., LTD	YB-AL001, 500g	Immunological adjuvant to enhance antigen immune response
BCA protein Concentration Test Kit	Fode Biotechnology Co., LTD	FD2001, 100 pieces per box	Total protein quantification to standardize sample loading
Chemiluminescence imaging system	Tanon	5200Multi	WB band imaging, gel and blot image collection and analysis
Color Pre-stained Protein Marker	Biolaibo Technology Co., LTD	BL005, 10-180kDa, 500μL	Protein molecular weight reference for electrophoresis experiments
Enhanced RNA Extraction Kit	Whole Golden Branch	ET111-01v2, 50 pieces per box	Extraction and purification of total RNA from tissues and cells
Enzyme-linked immunosorbent assay reader	U.S. Biotek Company	Epoch/BioTek	Quantitative detection of cytokines and protein contents in samples
FOXP3 primary antibody	Biyun Tian Biotechnology Co., LTD	AG5002, 50ul/piece	Targeted binding to FOXP3 protein for immunoassay
Gel Rapid Preparation Kit	Shanghai Yamei	PG212, 30 sets	Rapid preparation of separating gel and stacking gel for electrophoresis
masson Staining Kit	Solabao Biotechnology Co., LTD	G1340, 100 tablets	Specific staining for tissue collagen fibers and fibrosis observation
OVA	Beijing Boaotoda Technology Co., LTD.	OVA-10g, ≥98%	Classical allergen for establishing allergic animal models
primary Antibody Dilution solution	Biyun Tian Biotechnology Co., LTD	P0256-100ml	Dilute primary antibody, maintain activity and reduce non-specific binding
Rat IL-10ELISA Kit	Andy Bio	E-30649, 96T	Quantitative detection of rat IL-10 anti-inflammatory factor
Rat IL-4ELISA Kit	Andy Bio	E-30623, 96T	Quantitative detection of rat Th2 inflammatory factor IL-4
Reverse Transcription Kit	Baori Medical Technology Co., LTD	E047-01A	Reverse transcription of RNA into cDNA for subsequent gene analysis
Saturated Oil Red O Kit	Solabao Biotechnology Co., LTD	G1261, 50mL×2 bottle	Specific staining and detection of tissue lipid droplets
Shangyang Anti-Rabbit Secondary Antibody	Proteintech Biological Co., LTD	SA00001-2, 1mL	Bind to rabbit primary antibody for WB color reaction
TBST Buffer	Wuhan Sewell Biotechnology Co., LTD	G2150, 1L / bottle	Buffer for membrane washing and antibody dilution in Western blot
Transfer Electrophoresis System	Shanghai Jingcanshi Precision Machinery Co., Ltd.	PowerPac Basic	Used for protein electrophoresis and Western blot membrane transfer
Ultra-low temperature refrigerator	Zhongke Meiling	MDF-382	Low-temperature cryopreservation of reagents, protein samples and tissues
Vertical electrophoresis tank	BIO-RAD	JY-ZY5	Vertical electrophoretic separation of protein and nucleic acid samples