Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies

Escherichia coli (E. coli) is the primary causative agent of bovine mastitis. Currently, antibiotic therapy remains the cornerstone of mastitis treatment; necessitating the identification of alternative therapeutic options. This study employed in vitro cultured bovine bone marrow-derived macrophages...

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Main Authors: Xiaolin Yang, Xueqiang Li, Lili Guo, Pengfei Gong, Yinghong Qian, Shuangyi Zhang, Bo Liu, Wenrui Guo, Haixia Bao, Wei Mao
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Veterinary Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2025.1628028/full
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author Xiaolin Yang
Xiaolin Yang
Xueqiang Li
Xueqiang Li
Lili Guo
Lili Guo
Pengfei Gong
Pengfei Gong
Pengfei Gong
Yinghong Qian
Shuangyi Zhang
Shuangyi Zhang
Bo Liu
Bo Liu
Wenrui Guo
Wenrui Guo
Haixia Bao
Haixia Bao
Wei Mao
Wei Mao
author_facet Xiaolin Yang
Xiaolin Yang
Xueqiang Li
Xueqiang Li
Lili Guo
Lili Guo
Pengfei Gong
Pengfei Gong
Pengfei Gong
Yinghong Qian
Shuangyi Zhang
Shuangyi Zhang
Bo Liu
Bo Liu
Wenrui Guo
Wenrui Guo
Haixia Bao
Haixia Bao
Wei Mao
Wei Mao
author_sort Xiaolin Yang
collection DOAJ
description Escherichia coli (E. coli) is the primary causative agent of bovine mastitis. Currently, antibiotic therapy remains the cornerstone of mastitis treatment; necessitating the identification of alternative therapeutic options. This study employed in vitro cultured bovine bone marrow-derived macrophages (BMDMs) to systematically assess the potential of microsomal prostaglandin e synthase-1 (mPGES-1) inhibitors (MF63, MK886) and EP4 receptor inhibitor (Grapiprant) in modulating inflammatory responses and reducing tissue damage. Cells were pre-treated with mPGES-1 inhibitors and an EP4 receptor inhibitor before infection with E. coli. Following infection, extracellular bacteria were removed, and assays—including ELISA, Western blot, and qRT-PCR—were conducted to analyze inflammatory mediators, protein expression, and gene expression. E. coli infection significantly induced PGE₂ synthesis in BMDMs, which exacerbated the inflammatory response and tissue damage via NF-κB and MAPK signaling pathways, elevating TNF-α, IL-1β, IL-6, and IL-8. Treatment with MF63, MK886 and Grapiprant effectively reduced PGE₂ levels, inhibited NF-κB and MAPK signaling pathways, decreased inflammatory mediators, and enhanced macrophage bactericidal activity, thereby demonstrating potent anti-inflammatory and immunomodulatory effects. Moreover, inhibition of the mPGES-PGE₂-EP4 signaling pathway was found to reduce the expression of damage-associated molecular patterns (HMGB-1 and HABP-2), suggesting alleviation of E. coli-induced tissue damage. Based on the role of PGE₂ in mediating immune and inflammatory responses via the EP4 receptor, inhibiting the mPGES-1-PGE₂-EP4 signaling axis to reduce inflammation and tissue damage will facilitate further investigation into the regulatory mechanisms of the PGE₂ signaling axis in the pathogenesis of mastitis. This approach provides a theoretical foundation and experimental basis for the development of alternative anti-inflammatory therapies to replace antibiotics.
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spelling doaj-art-0e5ff18b95fc40a4bbcf90f577f5d7d42025-08-20T03:31:20ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-07-011210.3389/fvets.2025.16280281628028Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategiesXiaolin Yang0Xiaolin Yang1Xueqiang Li2Xueqiang Li3Lili Guo4Lili Guo5Pengfei Gong6Pengfei Gong7Pengfei Gong8Yinghong Qian9Shuangyi Zhang10Shuangyi Zhang11Bo Liu12Bo Liu13Wenrui Guo14Wenrui Guo15Haixia Bao16Haixia Bao17Wei Mao18Wei Mao19Key Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaInner Mongolia Bayannaoer City Municipal Center for Disease Control and Prevention, Bayannur, ChinaInner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaKey Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, ChinaLaboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, ChinaEscherichia coli (E. coli) is the primary causative agent of bovine mastitis. Currently, antibiotic therapy remains the cornerstone of mastitis treatment; necessitating the identification of alternative therapeutic options. This study employed in vitro cultured bovine bone marrow-derived macrophages (BMDMs) to systematically assess the potential of microsomal prostaglandin e synthase-1 (mPGES-1) inhibitors (MF63, MK886) and EP4 receptor inhibitor (Grapiprant) in modulating inflammatory responses and reducing tissue damage. Cells were pre-treated with mPGES-1 inhibitors and an EP4 receptor inhibitor before infection with E. coli. Following infection, extracellular bacteria were removed, and assays—including ELISA, Western blot, and qRT-PCR—were conducted to analyze inflammatory mediators, protein expression, and gene expression. E. coli infection significantly induced PGE₂ synthesis in BMDMs, which exacerbated the inflammatory response and tissue damage via NF-κB and MAPK signaling pathways, elevating TNF-α, IL-1β, IL-6, and IL-8. Treatment with MF63, MK886 and Grapiprant effectively reduced PGE₂ levels, inhibited NF-κB and MAPK signaling pathways, decreased inflammatory mediators, and enhanced macrophage bactericidal activity, thereby demonstrating potent anti-inflammatory and immunomodulatory effects. Moreover, inhibition of the mPGES-PGE₂-EP4 signaling pathway was found to reduce the expression of damage-associated molecular patterns (HMGB-1 and HABP-2), suggesting alleviation of E. coli-induced tissue damage. Based on the role of PGE₂ in mediating immune and inflammatory responses via the EP4 receptor, inhibiting the mPGES-1-PGE₂-EP4 signaling axis to reduce inflammation and tissue damage will facilitate further investigation into the regulatory mechanisms of the PGE₂ signaling axis in the pathogenesis of mastitis. This approach provides a theoretical foundation and experimental basis for the development of alternative anti-inflammatory therapies to replace antibiotics.https://www.frontiersin.org/articles/10.3389/fvets.2025.1628028/fullEscherichia colimastitisalternative treatments to antibioticsmPGES-1 inhibitorsEP4 receptor inhibitor
spellingShingle Xiaolin Yang
Xiaolin Yang
Xueqiang Li
Xueqiang Li
Lili Guo
Lili Guo
Pengfei Gong
Pengfei Gong
Pengfei Gong
Yinghong Qian
Shuangyi Zhang
Shuangyi Zhang
Bo Liu
Bo Liu
Wenrui Guo
Wenrui Guo
Haixia Bao
Haixia Bao
Wei Mao
Wei Mao
Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
Frontiers in Veterinary Science
Escherichia coli
mastitis
alternative treatments to antibiotics
mPGES-1 inhibitors
EP4 receptor inhibitor
title Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
title_full Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
title_fullStr Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
title_full_unstemmed Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
title_short Investigating the role of the mPGES-PGE₂-EP4 pathway in Escherichia coli-induced mastitis in dairy cows: insights for non-antibiotic therapeutic strategies
title_sort investigating the role of the mpges pge₂ ep4 pathway in escherichia coli induced mastitis in dairy cows insights for non antibiotic therapeutic strategies
topic Escherichia coli
mastitis
alternative treatments to antibiotics
mPGES-1 inhibitors
EP4 receptor inhibitor
url https://www.frontiersin.org/articles/10.3389/fvets.2025.1628028/full
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