Screening of key host targets and mechanistic assessment of berberine modulation of host responses to respiratory syncytial virus

Abstract

Objective

To screen potential host targets and investigate the molecular mechanism by which berberine regulates host responses to respiratory syncytial virus infection.

Methods

The structure and potential targets of berberine were retrieved and predicted using the PubChem, Swiss Target Prediction, and Pharm Mapper databases. Respiratory syncytial virus–related targets were screened from the GeneCards and Online Mendelian Inheritance in Man databases. Intersection targets were obtained using Venny 2.1.0. A protein–protein interaction network of the intersection targets was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins, and core targets were screened by performing topological analysis using Cytoscape. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed using the Database for Annotation, Visualization and Integrated Discovery. Molecular docking between berberine and the core targets was conducted using PyMol and AutoDock.

Results

In total, 299 berberine targets and 1196 respiratory syncytial virus targets were obtained, with 72 intersecting targets. Protein–protein interaction analysis identified 6 core targets, including epidermal growth factor receptor, Src, heat shock protein 90-alpha, albumin, signal transducer and activator of transcription 1, and caspase-3. Gene Ontology enrichment analysis indicated that the intersecting targets were involved in biological processes such as the phosphatidylinositol 3-kinase/protein kinase B signaling pathway, localized in cellular components including cytosol, and possessed molecular functions such as protein kinase activity. Kyoto Encyclopedia of Genes and Genomes analysis enriched 145 signaling pathways. Molecular docking results showed that the binding energies of berberine to the 8 core targets were all lower than −5 kcal/mol, presenting strong binding affinity.

Conclusion

Through network pharmacology and molecular docking, this study preliminarily suggests that berberine targets epidermal growth factor receptor, Src, heat shock protein 90-alpha, albumin, signal transducer and activator of transcription 1, and caspase-3 and regulates cell proliferation, inflammatory response, antiviral immunity, and cell apoptosis to modulate host response to respiratory syncytial virus infection. This study represents a purely bioinformatics prediction research, and its findings potentially provide a preliminary theoretical basis for elucidating the mechanism by which berberine regulates respiratory syncytial virus infection.

Introduction

Respiratory syncytial virus (RSV) is an enveloped negative-sense single-stranded RNA virus belonging to the Orthopneumovirus genus of the Pneumoviridae family. It can cause acute upper and lower respiratory tract infections (LRTIs) and is sometimes accompanied with extrapulmonary complications. The disease burden of RSV infection is extremely heavy, mainly affecting infants and older individuals aged >75 years. ¹ There are approximately 33 million new cases of acute LRTI each year, mainly occurring in children aged <5 years. High-risk populations, especially premature infants and individuals with underlying chronic lung disease, congenital heart disease, or impaired immune systems, are the most severely affected. ² In adults, the disease burden of RSV infection is comparable with that of influenza virus. ³ The pathological mechanism of RSV infection involves airway epithelial cell damage caused by viral replication and excessive immune inflammatory response. The clinical manifestations mainly include cough, wheezing, dyspnea, and other symptoms, and in severe cases, the condition can progress to respiratory failure.^4,5 At present, clinical treatment is mainly based on symptomatic support. Widespread use of the only available monoclonal antibodies (such as palivizumab) is limited owing to its high cost and the limited eligible population, whereas antiviral drugs such as ribavirin have limited efficacy and notable adverse effects. ⁶ Therefore, there is an urgent need to explore safe and effective anti-RSV drugs. ⁷

Berberine (chemical formula: C₂₀H₁₈NO₄⁺; molecular weight: 336.4 g/mol, chemical structure shown in Figure 1) is an isoquinoline alkaloid extracted from the Chinese herbal medicine Coptis chinensis and other Berberis plants, with a wide range of pharmacological activities.^8–10 Modern pharmacological studies have confirmed that berberine exerts multiple effects, including anticancer, antibacterial, antioxidative, antiapoptotic, and metabolic regulatory effects. ⁸ In addition, berberine has been reported to demonstrate antiviral activity, inhibiting infections caused by a variety of viruses, including RSV, human cytomegalovirus (HCMV), human papillomavirus (HPV), hepatitis C virus (HCV), and influenza virus. ¹¹ In addition, recent studies have confirmed that berberine and its derivatives exert direct antiviral activity against RSV in vitro. ¹⁰ Other studies have reported that berberine inhibits RSV replication and suppresses virus-induced p38 kinase activation. ⁹ However, the systematic host targets and overall regulatory mechanisms of berberine in RSV infection remain to be fully elucidated. The antiviral activity of berberine against several viruses makes it a promising candidate for the development of new antiviral drugs.

Figure 1.

Chemical structure of berberine.

Network pharmacology, based on the complex network relationship of “drug–target–disease,” enables the analysis of the multitarget and multipathway mechanism of drug action from a systems perspective, thereby overcoming the limitations of traditional single-target research. ¹² In this study, network pharmacology and molecular docking were used to screen potential host targets of berberine in the context of RSV infection, identify core targets, analyze biological functions and signaling pathways via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and verify the binding ability between berberine and core host targets. This study aimed to systematically clarify the molecular mechanism of berberine in regulating host responses to RSV infection and provide experimental clues and theoretical support for its application.

Materials and methods

Screening of berberine action targets

The two-dimensional (2D) structure and SMILES string of berberine were retrieved from the PubChem database (https://pubchem.ncbi.nlm.nih.gov/). ¹³ Potential targets were predicted using SwissTargetPrediction (http://www.swisstargetprediction.ch/) (2024 version, accessed in 2025) and PharmMapper (http://www.lilab-ecust.cn/pharmmapper/) (2017 version, accessed in 2025), with the species restricted to Homo sapiens for both databases.^14,15 For SwissTargetPrediction, the probability score was >0, no Top N limit was set, and all valid targets were retained. For PharmMapper, the platform's default Fit-score criteria were adopted, no custom threshold was set, and the top 300 ranked predicted targets were retained. ¹⁶ After merging the targets from the two databases, standardized mapping of gene symbols was performed using the UniProt database (human, accessed in 2025) (https://www.uniprot.org/) as the standard. ¹⁷ Duplicate targets, nonhuman targets, invalid targets, targets with missing values, and low-confidence entries were strictly excluded to screen out the potential action targets of berberine.

Screening of RSV targets

RSV-related targets were retrieved from the GeneCards (https://www.genecards.org) and Online Mendelian Inheritance in Man (OMIM; https://www.omim.org) databases using the keyword “respiratory syncytial virus.”^18,19

GeneCards database. The median relevance score was used as the cutoff to screen highly relevant targets; score thresholds of ≥5, ≥10, and ≥15 were also set for sensitivity analysis to validate target stability.^20–22

OMIM database. Duplicate targets were removed, and indirect or irrelevant disease-associated targets were excluded; only high-confidence candidate targets were retained.^16,19 Finally, targets from the two databases were merged and deduplicated to obtain the final RSV candidate targets.

Screening of intersection targets of berberine against RSV

The screened action targets of berberine and RSV were uploaded to the Venny 2.1.0 platform (https://bioinfogp.cnb.csic.es/tools/venny/) to obtain the intersection targets of the two, and a Venn diagram was constructed.

Construction and analysis of the protein–protein interaction (PPI) network

The intersection targets of berberine and RSV were imported into the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database (https://string-db.org) (version 12.0, accessed in 2026), with the species set to Homo sapiens. The interaction confidence threshold was set to ≥0.7, and all seven evidence types, including text mining, experiments, databases, co-expression, neighborhood, gene fusion, and co-occurrence, were included. A PPI network was then constructed. Network visualization and topological analysis were performed using Cytoscape v3.9.1. The CytoHubba plugin was used to screen hub genes by integrating four centrality algorithms: (a) degree; (b) maximal clique centrality (MCC); (c) betweenness; and (d) closeness centrality. The top 20 candidate targets ranked by each algorithm were selected, and the intersection of the four algorithm results was used to obtain the key core genes.^23,24

GO functional enrichment and KEGG pathway analysis

The intersection targets of berberine and RSV were imported into the Database for Annotation, Visualization and Integrated Discovery (DAVID) (https://david.ncifcrf.gov/) for GO and KEGG pathway functional enrichment analyses. The species was set to “Homo sapiens.” The Benjamini–Hochberg (BH) method was used for multiple testing correction to calculate the false discovery rate (FDR). Enriched terms with an adjusted q-value <0.05 were regarded as the screening criterion for significant enrichment.^25,26 The top 10 entries in GO function (including biological process (BP), cellular component (CC), and molecular function (MF)) were obtained, and the top 10 pathways in KEGG enrichment analysis were obtained. A GO function bubble chart and a KEGG enrichment bubble chart were drawn after visualization.

Molecular docking

The SDF structure of berberine was obtained from the PubChem small molecule database (https://pubchem.ncbi.nlm.nih.gov). After energy minimization and structural optimization, rotatable bonds were set using AutoDockTools 1.5.6, and the file was saved in the Protein Data Bank, Partial Charge, and Atom Type (PDBQT) format.

The crystal structures of core target proteins were retrieved from the Protein Data Bank (PDB) database (https://www.rcsb.org) with the PDB IDs, epidermal growth factor receptor (EGFR; 3p0y), Src (1o4j), heat shock protein 90-alpha (HSP90AA1; 2qfo), albumin (ALB; 6l4k), signal transducer and activator of transcription 1 (STAT1; 8d3f), and caspase-3 (CASP3; 3edq). PyMOL 2.3 was used to remove water molecules and original ligands. Subsequently, hydrogen addition as well as calculation and assignment of partial charges were performed using AutoDockTools 1.5.6. Molecular docking was performed using AutoDock Vina 1.1.2. The grid box parameters were set as follows: the grid size along the X/Y/Z axes was set to 40 with a grid spacing of 0.375 Å. The search exhaustiveness was set to the software default value of 8, and the number of output conformations (num_modes) was set at 10.^27,28 All other parameters remained at default values, and the built-in convergence criteria of the software were adopted.

Results

Acquisition of berberine action targets

Potential targets of berberine were predicted using the Swiss Target Prediction and PharmMapper databases, yielding 112 and 220 predicted targets, respectively. After merging and deduplication, a total of 299 standardized nonduplicate targets were obtained for subsequent analysis (Figure 2, Supplementary Table S1).

Figure 2.

Target network diagram of berberine.

Screening of action targets of berberine against RSV

RSV-related action targets were screened from the GeneCards and OMIM databases. After merging and deduplication, a total of 1196 action targets of RSV were screened. The targets of berberine and RSV were imported into Venny 2.1.0, and 72 intersecting targets were acquired. A Venn diagram was plotted accordingly (Figure 3). All intersecting targets were annotated with gene symbols and UniProt IDs (Supplementary Table S2).

Figure 3.

Venn diagram of intersection targets of berberine against respiratory syncytial virus.

PPI network and screening of key targets

The 72 overlapping targets of berberine against RSV were uploaded to the STRING database, with the species set to Homo sapiens and the minimum interaction confidence score set at 0.7. The results were exported in tab-separated values (TSV) format to construct the PPI network, which contained 72 nodes and 208 edges, with an average degree of 5.78. Network visualization was performed using Cytoscape v3.9.1 (Figure 4).

Figure 4.

PPI network of potential targets of berberine against respiratory syncytial virus.

PPI: protein–protein interaction.

Based on the CytoHubba plugin, four topological algorithms—degree, MCC, betweenness, and closeness—were combined to screen core targets. The top 20 candidate genes from each algorithm were selected (Figure 5), and their intersection yielded 8 key core genes: (a) EGFR; (b) SRC; (c) HSP90AA1; (d) ALB; (e) STAT1; (f) CASP3; (g) IGF1; and (h) PTGS2 (Table 1).

Figure 5.

Network diagram of core targets of berberine against respiratory syncytial virus based on four CytoHubba algorithms. (a) Degree; (b) MCC; (c) betweenness; (d) closeness; (e) Venn diagram of eight common hub proteins.

MCC: maximal clique centrality.

Table 1.

The topological characteristics of eight hub proteins.

Hub protein	Degree	MCC	Betweenness	Closeness
EGFR	28	2699	1014	45
SRC	24	2594	547	42
HSP90AA1	23	228	681	41
ALB	16	62	1093	38
STAT1	15	155	336	37
CASP3	14	53	549	38
IGF1	12	90	188	35
PTGS2	11	45	241	36

^MCC:maximal clique centrality; EGFR: epidermal growth factor receptor; HSP90AA1: heat shock protein 90-alpha; ALB: albumin; STAT1: signal transducer and activator of transcription 1; CASP3: caspase-3; IGF1: insulin-like growth factor 1; PTGS2: prostaglandin-endoperoxide synthase 2.

Sensitivity analysis was further conducted based on GeneCards relevance scores (cutoffs: ≥5, ≥10, ≥15, and median). EGFR, SRC, HSP90AA1, ALB, STAT1, and CASP3 were consistently identified across all thresholds; insulin-like growth factor 1 (IGF1) was stable at thresholds of ≥5, ≥10, and the median; prostaglandin-endoperoxide synthase 2 (PTGS2) was stable at thresholds of ≥10, ≥15, and the median. The above results confirmed that the six core targets, including EGFR, SRC, HSP90AA1, ALB, STAT1, and CASP3, exhibited excellent stability and high reliability across different thresholds.

GO pathway enrichment analysis of intersection targets

GO and KEGG enrichment analyses were performed on the intersecting targets of berberine and RSV to explore the regulatory mechanisms of berberine on host responses during RSV infection using the DAVID database. The GO enrichment results identified a total of 342 BPs, 50 CCs, and 81 MFs.

Among them, the BPs were mainly associated with the positive regulation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signal transduction, positive regulation of cell migration, negative regulation of apoptotic process, positive regulation of mitogen-activated protein kinase (MAPK) cascade, response to xenobiotic stimulus, positive regulation of gene expression, PI3K/Akt signal transduction, positive regulation of extracellular signal-regulated kinase 1 (ERK1) and extracellular signal-regulated kinase 2 (ERK2) cascade, positive regulation of cell population proliferation, and response to lipopolysaccharide.

The CCs primarily included cytosol, extracellular space, cytoplasm, membrane raft, extracellular region, receptor complex, extracellular exosome, ficolin-1-rich granule lumen, extracellular matrix, and focal adhesion.

The MFs mainly comprised protein kinase activity, kinase activity, endopeptidase activity, protein tyrosine kinase activity, adenosine triphosphate (ATP) binding, nucleotide binding, enzyme binding, identical protein binding, transferase activity, and transmembrane receptor protein tyrosine kinase activity (Figure 6).

Figure 6.

GO functional enrichment analysis of intersection targets. (a) Biological processes (BP); (b) cellular components (CC); (c) molecular functions (MF).

GO: Gene Ontology.

KEGG enrichment analysis

KEGG enrichment analysis results showed that a total of 145 signaling pathways were enriched, including pathways in cancer, lipid and atherosclerosis, fluid shear stress and atherosclerosis, advanced glycation end product (AGE)–receptor for advanced glycation end products (RAGE) signaling pathway in diabetic complications, proteoglycans in cancer, tumor necrosis factor (TNF) signaling pathway, Ras signaling pathway, PI3K/Akt signaling pathway, prostate cancer, vascular endothelial growth factor (VEGF) signaling pathway, and other signaling pathways (Figure 7).

Figure 7.

KEGG enrichment analysis.

KEGG: Kyoto Encyclopedia of Genes and Genomes.

Molecular docking

Berberine was docked with core targets (EGFR, Src, HSP90AA1, ALB, STAT1, and CASP3) involved in RSV infection. The results showed that berberine exhibited favorable binding affinity to all these core proteins, with binding energies for all lower than −5 kcal/mol. Specifically, the binding energies for the binding of berberine to EGFR, Src, HSP90AA1, ALB, STAT1, and CASP3 were −7.3, −6.8, −8.9, −10.1, −6.6, and −7.0 kcal/mol, respectively (Table 2). These strong interactions between berberine and core host proteins may underlie its regulatory effects on RSV infection–associated cellular processes, thereby contributing to its anti-RSV–related effects (Figure 8).

Table 2.

Molecular docking results of berberine with core targets (kcal/mol).

Target name	Protein data Bank ID	Binding energy (Kcal/mol)
EGFR	3p0y	−7.3
SRC	1o4j	−6.8
HSP90AA1	2qfo	−8.9
ALB	6l4k	−10.1
STAT1	8d3f	−6.6
CASP3	3edq	−7.0

ID: identification.

Figure 8.

Schematic diagram of molecular docking between berberine and core targets.

Discussion

RSV infection is a common clinical respiratory disease, which is particularly harmful to infants and immunocompromised populations. ²⁹ Berberine, a natural alkaloid with diverse pharmacological activities, has been shown to exert direct anti-RSV activity and modulate kinase signaling pathways during viral infection.^9,10 However, its anti-RSV mechanism remains unclear. In this study, network pharmacology and molecular docking technology were used to systematically explore the potential targets and signaling pathways of berberine in regulating host responses to RSV infection, providing novel ideas for clarifying its mechanism of action.

In this study, 299 potential action targets of berberine and 1196 related targets of RSV were first screened using databases. Seventy-two intersection targets were obtained using Venny platform analysis, indicating that these are key targets for berberine to exert anti-RSV effects.

PPI network analysis showed that four topological algorithms, including degree, MCC, betweenness, and closeness, were combined for hub target screening and sensitivity analysis based on GeneCards correlation score. Six pivotal core genes were finally obtained by intersection analysis, including EGFR, SRC, HSP90AA1, ALB, STAT1, and CASP3.

Among them, EGFR is an epidermal growth factor receptor involved in various BPs such as cell proliferation and differentiation. ³⁰ Studies have shown that RSV infection can activate the EGFR signaling pathway and promote viral replication. ³¹ Src is a nonreceptor tyrosine kinase that plays an important role in cell signal transduction and is closely related to the inflammatory response caused by viral infections. ³² HSP90AA1, as a heat shock protein, plays an important role in signal transduction and viral proliferation.^33,34 ALB is involved in drug transport and metabolism. ³⁵ In addition, existing literature has reported that ALB can exert indirect biological effects through immunomodulation, regulate host immune responses after viral infection, and enhance the body's antiviral immune capacity. ³⁶ HSP90AA1 and ALB may not act as direct antiviral targets and lack RSV specificity. Their identification as core targets reflects the multitarget pharmacological characteristics of berberine and represents an inherent feature of the multitarget analytical approach in network pharmacology. STAT1 is a member of the signal transducer and activator of transcription family, which plays an important role in antiviral immune response and can regulate the expression of antiviral factors such as interferons. ³⁷ CASP3 is a cysteine protease involved in the process of cell apoptosis, and viral infection often affects viral replication and release by regulating cell apoptosis. ³⁸ RSV reportedly inhibits CASP3 in the early stage of infection to facilitate viral replication, whereas it activates CASP3 in the late stage to promote viral release and inflammatory response. ³⁹ The functions of these core targets suggest that berberine alleviates RSV-induced host injury by regulating cell proliferation, inflammatory response, antiviral immune response, and cell apoptosis.

GO functional enrichment analysis revealed that the intersection targets of berberine against RSV were mainly involved in BPs such as positive regulation of PI3K/Akt signal transduction, positive regulation of cell migration, and negative regulation of apoptotic process; CCs such as cytosol, extracellular space, and membrane raft; and MFs such as protein kinase activity, ATP binding, and enzyme binding. The PI3K/Akt signaling pathway is an important intracellular signaling pathway involved in the regulation of cell survival, proliferation, apoptosis, and other processes, and a variety of viruses can promote their own replication by activating this pathway. ⁴⁰ Negative regulation of cellular apoptosis is closely associated with RSV infection. In the early stage of RSV infection, the PI3K/Akt signaling pathway is activated to negatively regulate the apoptotic process, delay host cell death, prolong cell survival time, and thereby create favorable conditions for viral replication. ⁴¹ These BPs and MFs further support that berberine may mitigate RSV-triggered host injury through regulation of cell signal transduction and metabolic processes.

KEGG pathway enrichment analysis revealed that the intersection targets of berberine against RSV were mainly enriched in pathways including those in cancer, the TNF signaling pathway, and Ras signaling pathway. Although some pathways seem to have no direct correlation with viral infection, many signaling pathways are abnormally activated during viral infection to meet the needs of viral replication and transmission. For example, the TNF signaling pathway is an important inflammatory signaling pathway, and RSV infection can activate this pathway, leading to the release of inflammatory factors, thereby causing lung inflammatory damage. ⁴² The Ras signaling pathway is involved in the regulation of cell proliferation and differentiation; it is also related to the viral replication cycle.^43,44 Berberine may inhibit viral replication and inflammatory response by regulating these signaling pathways, thereby reducing the damage caused by RSV infection.

Molecular docking results showed that berberine has good binding affinity for core targets, including EGFR, SRC, HSP90AA1, ALB, STAT1, and CASP3, with all binding energies less than −5 kcal/moll. This indicates that berberine can form strong potential binding interactions with these core targets, providing preliminary computational evidence for elucidating the molecular mechanism by which berberine regulates host responses to RSV infection.

This study is the first to adopt a systematic approach, which combines network pharmacology and molecular docking to comprehensively predict the core targets and molecular mechanisms of berberine in regulating host responses to RSV infection. By further identifying EGFR, SRC, HSP90AA1, ALB, STAT1, and CASP3 as key targets, this study suggests that berberine exerts a host-protective effect during RSV infection by modulating cell proliferation, inflammatory responses, host antiviral immune responses, and apoptosis. These findings complement and extend current research on the mechanisms by which berberine regulates host responses to RSV infection.^9,10,45 This work not only identifies potential therapeutic targets but also clarifies the multitarget and multipathway characteristics of berberine, providing new directions for the development of natural agents for RSV infection intervention. The integrated design of target screening, pathway enrichment, and molecular docking enhances the reliability of the predictions and provides a rational framework for subsequent experimental validation.

Limitations

This study has certain limitations. First, this research is a purely bioinformatics prediction analysis based on public databases, and all derived results are hypothetical without experimental verification. Since no in vitro cellular experiments or in vivo animal model validation were performed, the expression, interaction, and function of the predicted targets and signaling pathways cannot be confirmed. In addition, the dynamic changes and synergistic effects of multiple targets and pathways during RSV infection were not fully explored. Therefore, follow-up in vitro and in vivo RSV infection experiments will be conducted to determine the inhibitory effect of berberine on RSV viral load using TCID₅₀ and plaque assays and verify the alterations of core pathways and key targets such as EGFR phosphorylation, STAT1 activation, and CASP3 cleavage, to obtain experimental support for the proposed mechanisms.

Second, target screening in this study was only performed based on public databases, including GeneCards and OMIM, without integrating transcriptomic and proteomic data of RSV infection or experimentally validated RSV-host factor databases. In subsequent research, transcriptomic and proteomic data of RSV-infected cells and tissues from Gene Expression Omnibus (GEO) and ArrayExpress will be systematically integrated, combined with reported databases of RSV-specific host factors, to construct a more accurate disease target set. Meanwhile, in vitro cell experiments using RSV-infected HEp-2 cells will be applied to verify changes in the expression of core targets (e.g. STAT1 and CASP3) and the regulatory effect of berberine, thereby eliminating nonspecific signal interference at the experimental level. Third, this study mainly aimed to systematically elucidate the molecular mechanisms by which berberine alleviates RSV infection–induced injury in the host rather than exploring its direct antiviral effect. Moreover, the binding affinity between berberine and RSV viral proteins was not evaluated. Accordingly, this work is limited by the lack of experimental evidence on the direct interaction between berberine and RSV viral proteins as well as its in vitro anti-RSV activity. Further studies that aim to evaluate the binding between berberine and viral proteins and assess the vitro anti-RSV activity will be performed to clarify whether berberine exerts direct anti-RSV effects and provide sufficient experimental basis for its clinical application.

Finally, strict experimental validation of molecular docking was not conducted in this study; therefore, the relevant results are only theoretical predictions. The molecular docking analysis lacked rigorous validation designs such as redocking, positive controls, and negative controls, which is an inherent limitation of preliminary computational simulation research at this stage. Key experimental design elements mentioned above will be supplemented in follow-up studies to validate the docking results.

Conclusion

In this study, network pharmacology and molecular docking were used for preliminary exploration of the mechanism by which berberine regulates host responses to RSV infection. The results indicated that berberine may modulate host responses during RSV infection by acting on core host targets, including EGFR, SRC, HSP90AA1, ALB, STAT1, CASP3, and IGF1, regulating signaling pathways such as PI3K/Akt, TNF, and Ras and participating in the regulation of cell proliferation, inflammatory response, host antiviral immune response, and apoptosis. This study provides a theoretical basis for further research on berberine in the context of RSV infection, and all predicted host targets and mechanisms remain to be verified by in vitro and in vivo experiments. In subsequent studies, we will validate the expression and interactions between berberine and core host targets by performing vitro cell experiments and in vivo animal models as well as clarify the dose–effect and time–effect relationships of its activity in regulating host responses during RSV infection. The findings of this study can provide candidate targets for the development of berberine-based preparations targeting RSV infection and offer novel insights and research directions for the development of anti-RSV agents.