Angiotensin-(1-7)/MasR axis promotes migration of monocytes/macrophages with a regulatory phenotype to perform phagocytosis and efferocytosis
Isabella Zaidan, Luciana P. Tavares, Michelle A. Sugimoto, Kátia M. Lima, Graziele L. Negreiros-Lima, Lívia C.R. Teixeira, Thais C. Miranda, Bruno V.S. Valiate, Allysson Cramer, Juliana Priscila Vago, Gabriel H. Campolina-Silva, Jéssica A.M. Souza, Laís C. Grossi, Vanessa Pinho, Maria Jose Campagnole-Santos, Robson A.S. Santos, Mauro M. Teixeira, Izabela Galvão, Lirlândia P. Sousa
Isabella Zaidan, Luciana P. Tavares, Michelle A. Sugimoto, Kátia M. Lima, Graziele L. Negreiros-Lima, Lívia C.R. Teixeira, Thais C. Miranda, Bruno V.S. Valiate, Allysson Cramer, Juliana Priscila Vago, Gabriel H. Campolina-Silva, Jéssica A.M. Souza, Laís C. Grossi, Vanessa Pinho, Maria Jose Campagnole-Santos, Robson A.S. Santos, Mauro M. Teixeira, Izabela Galvão, Lirlândia P. Sousa
View: Text | PDF
Research Article Infectious disease Inflammation

Angiotensin-(1-7)/MasR axis promotes migration of monocytes/macrophages with a regulatory phenotype to perform phagocytosis and efferocytosis

Abstract

Nonphlogistic migration of macrophages contributes to the clearance of pathogens and apoptotic cells, a critical step for the resolution of inflammation and return to homeostasis. Angiotensin-(1-7) [Ang-(1-7)] is a heptapeptide of the renin-angiotensin system that acts through Mas receptor (MasR). Ang-(1-7) has recently emerged as a novel proresolving mediator, yet Ang-(1-7) resolution mechanisms are not fully determined. Herein, Ang-(1-7) stimulated migration of human and murine monocytes/macrophages in a MasR-, CCR2-, and MEK/ERK1/2–dependent manner. Pleural injection of Ang-(1-7) promoted nonphlogistic mononuclear cell influx alongside increased levels of CCL2, IL-10, and macrophage polarization toward a regulatory phenotype. Ang-(1-7) induction of CCL2 and mononuclear cell migration was also dependent on MasR and MEK/ERK. Of note, MasR was upregulated during the resolution phase of inflammation, and its pharmacological inhibition or genetic deficiency impaired mononuclear cell recruitment during self-resolving models of LPS pleurisy and E. coli peritonitis. Inhibition/absence of MasR was associated with reduced CCL2 levels, impaired phagocytosis of bacteria, efferocytosis, and delayed resolution of inflammation. In summary, we have uncovered a potentially novel proresolving feature of Ang-(1-7), namely the recruitment of mononuclear cells favoring efferocytosis, phagocytosis, and resolution of inflammation. Mechanistically, cell migration was dependent on MasR, CCR2, and the MEK/ERK pathway.

Authors

Isabella Zaidan, Luciana P. Tavares, Michelle A. Sugimoto, Kátia M. Lima, Graziele L. Negreiros-Lima, Lívia C.R. Teixeira, Thais C. Miranda, Bruno V.S. Valiate, Allysson Cramer, Juliana Priscila Vago, Gabriel H. Campolina-Silva, Jéssica A.M. Souza, Laís C. Grossi, Vanessa Pinho, Maria Jose Campagnole-Santos, Robson A.S. Santos, Mauro M. Teixeira, Izabela Galvão, Lirlândia P. Sousa

×

Figure 2

Ang-(1-7)–induced macrophage migration is dependent on MasR and associated with CCR2 and the MEK/ERK1/2 pathway.

Options: View larger image (or click on image) Download as PowerPoint
Ang-(1-7)–induced macrophage migration is dependent on MasR and associat...
(A) RAW 264.7 cells were pretreated with the antagonist of MasR (A779, 1 μM) 1 hour before performing chemotaxis assays using Ang-(1-7) (100 nM) as the chemoattractant. (B) BMDMs from WT and MasR–/– mice were next used to assess cell chemotaxis toward Ang-(1-7). (C) Graph shows the production of CCL2 by WT BMDMs 4 hours post–Ang-(1-7) exposure. (D) RAW 264.7 cells were pretreated with CCR2 antagonist (RS504393 10 μM) 1 hour before performing chemotaxis assays using Ang-(1-7) (100 nM) as the chemoattractant. (E) BMDMs from WT and CCR2–/– mice were used to assess cell chemotaxis toward Ang-(1-7), and (F) a chemotaxis assay toward CCL2 (0.1 and 1 ng/mL) was performed using BMDMs from WT and MasR–/– mice. (G) RAW 264.7 cells were pretreated for 1 hour with inhibitors of the MEK/ERK pathway (selumetinib and U0126, 10 and 15 μM, respectively), and chemotaxis assays using Ang-(1-7) (100 nM) as the chemoattractant were performed. (H) Western blot analyses for p-ERK1/2 were performed after incubation of RAW 264.7 cells with the abovementioned inhibitors followed by Ang-(1-7) treatment. β-Actin was used as a protein load control. (I) Quantification of blot bands was performed using ImageJ (NIH). Data are presented as mean ± SEM, * for P < 0.05, ** for P < 0.01, and *** for P < 0.001 when compared with the control group by 1-way ANOVA (A, B, and D–I) or 2-tailed t test (C). Data are representative of 3 independent experiments performed in biological triplicates or quadruplicates (n = 3–4). CM, control medium; UT, untreated cells.