Abstract
The role of protease-activated receptor (PAR)4 in thrombin-induced platelet aggregation has been studied, and PAR4 blockade is thought to be useful as a new and promising approach in antiplatelet therapy in humans. In recent years, studies have been conducted to clarify the role of PAR4 in the host defense against invading microorganisms and pathogen-induced inflammation; however, to date, the role of PAR4 in mediating the LPS-induced inflammatory repertoire in macrophages remains to be elucidated. Here, we investigated the effects of the synthetic PAR4 agonist peptide (PAR4-AP) AYPGKF-NH2 on the phagocytosis of zymosan-FITC particles; NO, ROS, and iNOS expression; and cytokine production in C57/BL6 macrophages cocultured with PAR4-AP/LPS. The PAR4-AP impaired LPS-induced and basal phagocytosis, which was restored by pharmacological PAR4 blockade. Coincubation with the PAR4-AP/LPS enhanced NO and ROS production and iNOS expression; decreased IL-10, but not TNF-α, in the culture supernatant; and increased translocation of the p65 subunit of the proinflammatory gene transcription factor NF-κ-B. Our results provide evidence for a complex mechanism and new approach by which PAR4 mediates the macrophage response triggered by LPS through counter-regulating the phagocytic activity of macrophages and innate response mechanisms implicated in the killing of invading pathogens.
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Acknowledgments
This work was supported by Fundacao de Amparo a Pesquisa de Minas Gerais (FAPEMIG/Brazil, grant number PPM-00593-16). AB was a graduate student fellow from CNPq/Brazil, and MTPL is a senior research fellow from CNPq/Brazil.
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AB conceived and conducted experiments, analyzed the data, and wrote the manuscript. KMF and DGM conducted experiments and analyzed the data. PFP and MTPL analyzed the data and contributed new reagents and analytical tools. AK supervised, conceived the experiments, designed research, analyzed the data, and wrote the manuscript. All authors read and approved the manuscript and declare that all data were generated in-house and that no paper mill was used.
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Barra, A., Freitas, K.M., Marconato, D.G. et al. Protease-activated receptor 4 plays a role in lipopolysaccharide-induced inflammatory mechanisms in murine macrophages. Naunyn-Schmiedeberg's Arch Pharmacol 394, 853–862 (2021). https://doi.org/10.1007/s00210-020-02014-w
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DOI: https://doi.org/10.1007/s00210-020-02014-w