Ayslan Barra1, Amanda Ferreira Brasil1, Thaís Lemos Ferreira2, Weslley Fernandes-Braga3, Danielle Gomes Marconato4, Priscila Faria-Pinto4, Jacqueline Isaura Alvarez-Leite3, Luciano Dos Santos Aggum Capettini2, André Klein5. 1. Laboratory of Pain and Inflammation, Department of Pharmacology, Institute of Biological Sciences (ICB), Federal University of Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627, Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil. 2. Laboratory of Vascular Biology, ICB/UFMG, Belo Horizonte, Minas Gerais, Brazil. 3. Laboratory of Atherosclerosis and Nutritional Biochemistry (LABIN-UFMG), Department of Biochemistry and Immunology, ICB/UFMG, Belo Horizonte, Minas Gerais, Brazil. 4. Department of Biochemistry, Federal University of Juiz de Fora (UFJF), Juiz de Fora, Minas Gerais, Brazil. 5. Laboratory of Pain and Inflammation, Department of Pharmacology, Institute of Biological Sciences (ICB), Federal University of Minas Gerais (UFMG), Av. Presidente Antônio Carlos, 6627, Pampulha, Belo Horizonte, Minas Gerais, 31270-901, Brazil. klein@ufmg.br.
Abstract
OBJECTIVE: This study was conducted to investigate the effects of the synthetic PAR2 agonist peptide (PAR2-AP) SLIGRL-NH2 on LPS-induced inflammatory mechanisms in peritoneal macrophages. METHODS: Peritoneal macrophages obtained from C57BL/6 mice were incubated with PAR2-AP and/or LPS, and the phagocytosis of zymosan fluorescein isothiocyanate (FITC) particles; nitric oxide (NO), reactive oxygen species (ROS), and cytokine production; and inducible NO synthase (iNOS) expression in macrophages co-cultured with PAR-2-AP/LPS were evaluated. RESULTS: Co-incubation of macrophages with PAR2AP (30 µM)/LPS (100 ng/mL) enhanced LPS-induced phagocytosis; production of NO, ROS, and the pro-inflammatory cytokines interleukin (IL)-1β, tumour necrosis factor (TNF)-α, IL-6, and C-C motif chemokine ligand (CCL)2; and iNOS expression and impaired the release of the anti-inflammatory cytokine IL-10 after 4 h of co-stimulation. In addition, PAR2AP increased the LPS-induced translocation of the p65 subunit of the pro-inflammatory transcription factor nuclear factor kappa B (NF-κB) and reduced the expression of inhibitor of NF-κB. CONCLUSION: This study provides evidence of a role for PAR2 in macrophage response triggered by LPS enhancing the phagocytic activity and NO, ROS, and cytokine production, resulting in the initial and adequate macrophage response required for their innate response mechanisms.
OBJECTIVE: This study was conducted to investigate the effects of the synthetic PAR2 agonist peptide (PAR2-AP) SLIGRL-NH2 on LPS-induced inflammatory mechanisms in peritoneal macrophages. METHODS: Peritoneal macrophages obtained from C57BL/6 mice were incubated with PAR2-AP and/or LPS, and the phagocytosis of zymosan fluorescein isothiocyanate (FITC) particles; nitric oxide (NO), reactive oxygen species (ROS), and cytokine production; and inducible NO synthase (iNOS) expression in macrophages co-cultured with PAR-2-AP/LPS were evaluated. RESULTS: Co-incubation of macrophages with PAR2AP (30 µM)/LPS (100 ng/mL) enhanced LPS-induced phagocytosis; production of NO, ROS, and the pro-inflammatory cytokines interleukin (IL)-1β, tumour necrosis factor (TNF)-α, IL-6, and C-C motif chemokine ligand (CCL)2; and iNOS expression and impaired the release of the anti-inflammatory cytokine IL-10 after 4 h of co-stimulation. In addition, PAR2AP increased the LPS-induced translocation of the p65 subunit of the pro-inflammatory transcription factor nuclear factor kappa B (NF-κB) and reduced the expression of inhibitor of NF-κB. CONCLUSION: This study provides evidence of a role for PAR2 in macrophage response triggered by LPS enhancing the phagocytic activity and NO, ROS, and cytokine production, resulting in the initial and adequate macrophage response required for their innate response mechanisms.
Authors: Mark N Adams; Rithwik Ramachandran; Mei-Kwan Yau; Jacky Y Suen; David P Fairlie; Morley D Hollenberg; John D Hooper Journal: Pharmacol Ther Date: 2011-01-26 Impact factor: 12.310
Authors: Martin Steinhoff; Jörg Buddenkotte; Victoria Shpacovitch; Anke Rattenholl; Corinna Moormann; Nathalie Vergnolle; Thomas A Luger; Morley D Hollenberg Journal: Endocr Rev Date: 2005-02 Impact factor: 19.871
Authors: Victoria M Shpacovitch; Micha Feld; Dirk Holzinger; Makiko Kido; Morley D Hollenberg; Francesca Levi-Schaffer; Nathalie Vergnolle; Stephan Ludwig; Johannes Roth; Thomas Luger; Martin Steinhoff Journal: Immunology Date: 2011-04-19 Impact factor: 7.397