Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation

Abstract Background Neutrophil Extracellular Traps (NETs) are closely related to the progression of inflammation in Chronic Obstructive Pulmonary Disease (COPD). Erythromycin (EM) has been shown to inhibit inflammation in COPD, but its molecular mechanisms is still unclear. The aim of our study is i...

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Main Authors: Nan Ma, Xiao Na Liang, Quan Fang Chen, Mei Hua Li, Guang Sheng Pei, Xiao Fei Yi, Li Yan Guo, Fu Gang Chen, Zhi Yi He
Format: Article
Language:English
Published: BMC 2025-04-01
Series:Respiratory Research
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Online Access:https://doi.org/10.1186/s12931-025-03226-5
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author Nan Ma
Xiao Na Liang
Quan Fang Chen
Mei Hua Li
Guang Sheng Pei
Xiao Fei Yi
Li Yan Guo
Fu Gang Chen
Zhi Yi He
author_facet Nan Ma
Xiao Na Liang
Quan Fang Chen
Mei Hua Li
Guang Sheng Pei
Xiao Fei Yi
Li Yan Guo
Fu Gang Chen
Zhi Yi He
author_sort Nan Ma
collection DOAJ
description Abstract Background Neutrophil Extracellular Traps (NETs) are closely related to the progression of inflammation in Chronic Obstructive Pulmonary Disease (COPD). Erythromycin (EM) has been shown to inhibit inflammation in COPD, but its molecular mechanisms is still unclear. The aim of our study is investigate the molecular mechanisms of EM's anti-inflammatory effects in NETs-induced inflammation. Methods Transcriptomics and proteomics data were obtained from U937 cells treated with NETs and EM. Differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were identified using R software. Pathway enrichment analyses, were employed to identify inflammation-related pathways. Cytoscape were utilized to construct network of hub targets regulated by EM which related with oxidative stress and inflammation. Additionally, Cytoscape and STRING were used to construct protein–protein interaction (PPI) network of key targets regulated by EM. The expression levels of key targets were further confirmed through WB and PCR experiments. Results Both transcriptomics and proteomics indicate that EM decrease NETs –induced AKT1 expression. Enrichment analysis of DEGs and DEPs reveal multiple common pathways involved in EM’s regulation inflammation, including the PI3K/AKT pathway, response to oxidative stress, IKK/NF-κB signaling and PTEN signaling pathway. Nine key targets in PI3K/AKT-related inflammatory pathways regulated by EM and ten targets of EM-regulated oxidative stress were identified. WB and PCR results confirmed that EM reversing the NETs-induced inflammation by modulating the activity of these targets. Furthermore, clinical samples and vitro experiments confirm that EM alleviates NETs-induced glucocorticoid resistance via inhibiting PI3K/AKT, thereby repressing inflammation. Conclusions Our study provides a comprehensive proteogenomic characterization of how EM alleviates NET-related inflammation, and identify PI3K/AKT play a critical role in the mechanism by which EM inhibits inflammation.
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spelling doaj-art-2977cbfb2f094e5eab26fbfa8ec31f992025-08-20T03:18:28ZengBMCRespiratory Research1465-993X2025-04-0126112510.1186/s12931-025-03226-5Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammationNan Ma0Xiao Na Liang1Quan Fang Chen2Mei Hua Li3Guang Sheng Pei4Xiao Fei Yi5Li Yan Guo6Fu Gang Chen7Zhi Yi He8Department of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityDepartment of Respiratory Medicine, First Affiliated Hospital of Guangxi Medical UniversityAbstract Background Neutrophil Extracellular Traps (NETs) are closely related to the progression of inflammation in Chronic Obstructive Pulmonary Disease (COPD). Erythromycin (EM) has been shown to inhibit inflammation in COPD, but its molecular mechanisms is still unclear. The aim of our study is investigate the molecular mechanisms of EM's anti-inflammatory effects in NETs-induced inflammation. Methods Transcriptomics and proteomics data were obtained from U937 cells treated with NETs and EM. Differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) were identified using R software. Pathway enrichment analyses, were employed to identify inflammation-related pathways. Cytoscape were utilized to construct network of hub targets regulated by EM which related with oxidative stress and inflammation. Additionally, Cytoscape and STRING were used to construct protein–protein interaction (PPI) network of key targets regulated by EM. The expression levels of key targets were further confirmed through WB and PCR experiments. Results Both transcriptomics and proteomics indicate that EM decrease NETs –induced AKT1 expression. Enrichment analysis of DEGs and DEPs reveal multiple common pathways involved in EM’s regulation inflammation, including the PI3K/AKT pathway, response to oxidative stress, IKK/NF-κB signaling and PTEN signaling pathway. Nine key targets in PI3K/AKT-related inflammatory pathways regulated by EM and ten targets of EM-regulated oxidative stress were identified. WB and PCR results confirmed that EM reversing the NETs-induced inflammation by modulating the activity of these targets. Furthermore, clinical samples and vitro experiments confirm that EM alleviates NETs-induced glucocorticoid resistance via inhibiting PI3K/AKT, thereby repressing inflammation. Conclusions Our study provides a comprehensive proteogenomic characterization of how EM alleviates NET-related inflammation, and identify PI3K/AKT play a critical role in the mechanism by which EM inhibits inflammation.https://doi.org/10.1186/s12931-025-03226-5ProteogenomicErythromycinNETsInflammationPI3K/AKT
spellingShingle Nan Ma
Xiao Na Liang
Quan Fang Chen
Mei Hua Li
Guang Sheng Pei
Xiao Fei Yi
Li Yan Guo
Fu Gang Chen
Zhi Yi He
Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
Respiratory Research
Proteogenomic
Erythromycin
NETs
Inflammation
PI3K/AKT
title Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
title_full Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
title_fullStr Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
title_full_unstemmed Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
title_short Proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps-induced inflammation
title_sort proteogenomic verifies targets underlying erythromycin alleviate neutrophil extracellular traps induced inflammation
topic Proteogenomic
Erythromycin
NETs
Inflammation
PI3K/AKT
url https://doi.org/10.1186/s12931-025-03226-5
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