Yang Xiaolin, Li Xueqiang, Guo Lili, Gong Pengfei, Qian Yinghong, Zhang Shuangyi, Liu Bo, Guo Wenrui, Bao Haixia, Mao Wei
Key Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease, Ministry of Agriculture, Inner Mongolia Agricultural University, Hohhot, China.
Laboratory of Veterinary Clinical Pharmacology, College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China.
Front Vet Sci. 2025 Jul 1;12:1628028. doi: 10.3389/fvets.2025.1628028. eCollection 2025.
() 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 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 . 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. infection significantly induced PGE₂ synthesis in BMDMs, which exacerbated the inflammatory response and tissue damage via NF-κB and MAPK signaling pathways, elevating TNF-, 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 -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.
（）是牛乳腺炎的主要病原体。目前，抗生素治疗仍然是乳腺炎治疗的基石；因此需要确定替代治疗方案。本研究使用培养的牛骨髓来源巨噬细胞（BMDMs）系统评估微粒体前列腺素E合酶-1（mPGES-1）抑制剂（MF63、MK886）和EP4受体抑制剂（格拉普兰特）在调节炎症反应和减少组织损伤方面的潜力。在用（）感染之前，细胞先用mPGES-1抑制剂和EP4受体抑制剂进行预处理。感染后，去除细胞外细菌，并进行包括酶联免疫吸附测定（ELISA）、蛋白质印迹法和定量逆转录聚合酶链反应（qRT-PCR）在内的检测，以分析炎症介质、蛋白质表达和基因表达。（）感染显著诱导BMDMs中PGE₂的合成，其通过核因子κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）信号通路加剧炎症反应和组织损伤，升高肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和白细胞介素-8（IL-8）。用MF63、MK886和格拉普兰特治疗可有效降低PGE₂水平，抑制NF-κB和MAPK信号通路，减少炎症介质，并增强巨噬细胞杀菌活性，从而显示出强大的抗炎和免疫调节作用。此外发现，抑制mPGES-PGE₂-EP4信号通路可降低损伤相关分子模式（高迁移率族蛋白B1（HMGB-1）和透明质酸结合蛋白-（HABP-2））的表达，提示减轻（）诱导的组织损伤。基于PGE₂通过EP4受体介导免疫和炎症反应的作用，抑制mPGES-1-PGE₂-EP4信号轴以减少炎症和组织损伤将有助于进一步研究PGE₂信号轴在乳腺炎发病机制中的调控机制。这种方法为开发替代抗生素的抗炎治疗方法提供了理论基础和实验依据。
