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Salt increases monocyte CCR2 expression and inflammatory responses in humans
Eliane F.E. Wenstedt, … , Liffert Vogt, Jan Van den Bossche
Eliane F.E. Wenstedt, … , Liffert Vogt, Jan Van den Bossche
Published November 1, 2019
Citation Information: JCI Insight. 2019;4(21):e130508. https://doi.org/10.1172/jci.insight.130508.
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Research Article Clinical trials Inflammation

Salt increases monocyte CCR2 expression and inflammatory responses in humans

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Abstract

Inflammation may play a role in the link between high salt intake and its deleterious consequences. However, it is unknown whether salt can induce proinflammatory priming of monocytes and macrophages in humans. We investigated the effects of salt on monocytes and macrophages in vitro and in vivo by performing a randomized crossover trial in which 11 healthy human subjects adhered to a 2-week low-salt and high-salt diet. We demonstrate that salt increases monocyte expression of CCR2, a chemokine receptor that mediates monocyte infiltration in inflammatory diseases. In line with this, we show a salt-induced increase of plasma MCP-1, transendothelial migration of monocytes, and skin macrophage density after high-salt diet. Macrophages demonstrate signs of an increased proinflammatory phenotype after salt exposure, as represented by boosted LPS-induced cytokine secretion of IL-6, TNF, and IL-10 in vitro, and by increased HLA-DR expression and decreased CD206 expression on skin macrophages after high-salt diet. Taken together, our data open up the possibility for inflammatory monocyte and macrophage responses as potential contributors to the deleterious effects of high salt intake.

Authors

Eliane F.E. Wenstedt, Sanne G.S. Verberk, Jeffrey Kroon, Annette E. Neele, Jeroen Baardman, Nike Claessen, Özge T. Pasaoglu, Emma Rademaker, Esmee M. Schrooten, Rosa D. Wouda, Menno P.J. de Winther, Jan Aten, Liffert Vogt, Jan Van den Bossche

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Figure 6

High salt (HS) boosts LPS-induced cytokine secretion in macrophages in vitro but does not affect the activation of M(IL-4).

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High salt (HS) boosts LPS-induced cytokine secretion in macrophages in v...
(A) Schematic overview of in vitro LPS stimulation of macrophages. Monocytes of healthy volunteers were cultured in the presence of M-CSF for 7 days to differentiate into mature macrophages and stimulated for 24 h with LPS in the presence of normal salt (NS) concentrations present in IMDM medium (NS; [Na+] = 139 mM), HS ([Na+] = 179 mM; i.e., IMDM medium supplemented with an additional 40 mM NaCl), or 80 mM urea as tonicity control. (B) IL-6, IL-10, IL-12, and TNF were quantified by ELISA. (C) Schematic overview of in vitro IL-4 stimulation of macrophages. (D) Differentially treated macrophages were stained with antibodies against the M2 surface markers CD206 and CD200R, or isotype control, followed by flow cytometric analysis. Representative histogram graphs and corresponding surface expression quantifications (ΔMFI = [MFI]positive staining – [MFI]isotype staining) are presented. (E) Gene expression of IL-4–induced M2 marker genes. The fold inductions of indicated marker genes are shown relative to the expression in untreated macrophages (= 1). M-CSF, macrophage CSF; NS, ([Na+] = 139 mM); HS, ([Na+] = 179 mM); MFI, median fluorescence intensity. Values represent mean ± SEM of at least n = 3 healthy male donors. n = 3 (1 outlier excluded for TNF for B, n = 6 for D, n = 3 for E). Data tested using 1-way ANOVA. *P < 0.05; **P < 0.01; ***P < 0.001.

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