Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway

ABSTRACT Berberine exhibits antitumor, antibacterial, and antiviral properties. This study explored its therapeutic potential and molecular mechanism in respiratory syncytial virus (RSV)-induced pediatric bronchiolitis (PB). BEAS-2B cells were infected with RSV (multiplicity of infection = 0.5, 1.0,...

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Main Authors: Yang Jiao, Rui Yang
Format: Article
Language:English
Published: American Society for Microbiology 2025-08-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.00900-25
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author Yang Jiao
Rui Yang
author_facet Yang Jiao
Rui Yang
author_sort Yang Jiao
collection DOAJ
description ABSTRACT Berberine exhibits antitumor, antibacterial, and antiviral properties. This study explored its therapeutic potential and molecular mechanism in respiratory syncytial virus (RSV)-induced pediatric bronchiolitis (PB). BEAS-2B cells were infected with RSV (multiplicity of infection = 0.5, 1.0, or 5.0) to establish a PB cell model. Quantitative reverse transcription PCR (qRT-PCR) quantified RSV, HMGB1, and fibrosis-related genes. MTT assay assessed cell viability, and plaque assay measured RSV titers. Western blot, immunohistochemistry (IHC), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) evaluated HMGB1 and key markers of inflammation, pyroptosis, epithelial-mesenchymal transition (EMT), and fibrosis. Flow cytometry analyzed apoptosis, while electron microscopy visualized RSV-induced pyroptotic and apoptotic changes. A PB mouse model was established by administering RSV (6.8 × 10⁶ PFU) via nasal drip in BALB/c mice. Mice were divided into six groups (n = 5 per group): blank, RSV, RSV + dimethyl sulfoxide (DMSO), RSV + berberine low dose, RSV + berberine medium dose and RSV + berberine high dose. HE staining examined lung pathology, while Western blot, IHC, ELISA, and immunofluorescence quantified HMGB1 and markers of inflammation, pyroptosis, EMT, and fibrosis. Berberine significantly reduced RSV replication, alleviated lung inflammation and fibrosis, and suppressed the HMGB1/TLR4/NF-κB pathway in both RSV-infected BEAS-2B cells and PB mice, as indicated by the downregulation of HMGB1, TLR4, p-IκBα, and p-p65. Berberine also inhibited pyroptosis (NLRP3, ASC, cleaved caspase-1, caspase-11, and GSDMD-N), EMT (Snail and N-cadherin), and fibrosis (CoL1A1, α-SMA, fibronectin, and ACTA2). Electron microscopy confirmed that RSV-induced cellular damage, including mitochondrial swelling and chromatin condensation, was alleviated by berberine treatment. These protective effects were reversed by HMGB1 overexpression and enhanced by TLR4 inhibition, indicating that berberine exerted its effects via the HMGB1/TLR4/NF-κB signaling pathway. Overall, berberine attenuated RSV-induced PB by suppressing the HMGB1/TLR4/NF-κB pathway, reducing inflammation, pyroptosis, EMT, and fibrosis in lung tissues and epithelial cells, which may be a promising remedy for RSV-induced PB.IMPORTANCEPediatric bronchiolitis caused by RSV remains a major global health challenge with limited treatment options. This study highlights berberine as a promising therapeutic agent capable of reducing RSV replication and associated lung injury. By targeting the HMGB1/TLR4/NF-κB pathway, berberine effectively attenuates inflammation, pyroptosis, EMT, and fibrosis. These findings provide a novel mechanistic insight and offer potential for the development of targeted therapies against RSV-induced bronchiolitis.
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spelling doaj-art-42f74958fe2c4f70bfe922db9f5d581b2025-08-20T03:44:01ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-08-0113810.1128/spectrum.00900-25Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathwayYang Jiao0Rui Yang1Department of Pediatrics, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, ChinaDepartment of Pediatrics, Hebei Reproductive Health Hospital, Hebei Women and Children's Health Hospital, Shijiazhuang, Hebei, ChinaABSTRACT Berberine exhibits antitumor, antibacterial, and antiviral properties. This study explored its therapeutic potential and molecular mechanism in respiratory syncytial virus (RSV)-induced pediatric bronchiolitis (PB). BEAS-2B cells were infected with RSV (multiplicity of infection = 0.5, 1.0, or 5.0) to establish a PB cell model. Quantitative reverse transcription PCR (qRT-PCR) quantified RSV, HMGB1, and fibrosis-related genes. MTT assay assessed cell viability, and plaque assay measured RSV titers. Western blot, immunohistochemistry (IHC), immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) evaluated HMGB1 and key markers of inflammation, pyroptosis, epithelial-mesenchymal transition (EMT), and fibrosis. Flow cytometry analyzed apoptosis, while electron microscopy visualized RSV-induced pyroptotic and apoptotic changes. A PB mouse model was established by administering RSV (6.8 × 10⁶ PFU) via nasal drip in BALB/c mice. Mice were divided into six groups (n = 5 per group): blank, RSV, RSV + dimethyl sulfoxide (DMSO), RSV + berberine low dose, RSV + berberine medium dose and RSV + berberine high dose. HE staining examined lung pathology, while Western blot, IHC, ELISA, and immunofluorescence quantified HMGB1 and markers of inflammation, pyroptosis, EMT, and fibrosis. Berberine significantly reduced RSV replication, alleviated lung inflammation and fibrosis, and suppressed the HMGB1/TLR4/NF-κB pathway in both RSV-infected BEAS-2B cells and PB mice, as indicated by the downregulation of HMGB1, TLR4, p-IκBα, and p-p65. Berberine also inhibited pyroptosis (NLRP3, ASC, cleaved caspase-1, caspase-11, and GSDMD-N), EMT (Snail and N-cadherin), and fibrosis (CoL1A1, α-SMA, fibronectin, and ACTA2). Electron microscopy confirmed that RSV-induced cellular damage, including mitochondrial swelling and chromatin condensation, was alleviated by berberine treatment. These protective effects were reversed by HMGB1 overexpression and enhanced by TLR4 inhibition, indicating that berberine exerted its effects via the HMGB1/TLR4/NF-κB signaling pathway. Overall, berberine attenuated RSV-induced PB by suppressing the HMGB1/TLR4/NF-κB pathway, reducing inflammation, pyroptosis, EMT, and fibrosis in lung tissues and epithelial cells, which may be a promising remedy for RSV-induced PB.IMPORTANCEPediatric bronchiolitis caused by RSV remains a major global health challenge with limited treatment options. This study highlights berberine as a promising therapeutic agent capable of reducing RSV replication and associated lung injury. By targeting the HMGB1/TLR4/NF-κB pathway, berberine effectively attenuates inflammation, pyroptosis, EMT, and fibrosis. These findings provide a novel mechanistic insight and offer potential for the development of targeted therapies against RSV-induced bronchiolitis.https://journals.asm.org/doi/10.1128/spectrum.00900-25respiratory syncytial viruspediatric bronchiolitisberberineinflammationepithelial-mesenchymal transformation
spellingShingle Yang Jiao
Rui Yang
Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
Microbiology Spectrum
respiratory syncytial virus
pediatric bronchiolitis
berberine
inflammation
epithelial-mesenchymal transformation
title Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
title_full Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
title_fullStr Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
title_full_unstemmed Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
title_short Berberine alleviates respiratory syncytial virus (RSV)-induced pediatric bronchiolitis and fibrosis via suppressing the HMGB1/TLR4/NF-κB pathway
title_sort berberine alleviates respiratory syncytial virus rsv induced pediatric bronchiolitis and fibrosis via suppressing the hmgb1 tlr4 nf κb pathway
topic respiratory syncytial virus
pediatric bronchiolitis
berberine
inflammation
epithelial-mesenchymal transformation
url https://journals.asm.org/doi/10.1128/spectrum.00900-25
work_keys_str_mv AT yangjiao berberinealleviatesrespiratorysyncytialvirusrsvinducedpediatricbronchiolitisandfibrosisviasuppressingthehmgb1tlr4nfkbpathway
AT ruiyang berberinealleviatesrespiratorysyncytialvirusrsvinducedpediatricbronchiolitisandfibrosisviasuppressingthehmgb1tlr4nfkbpathway