Journal Article

Deletion of Rac1GTPase in the Myeloid Lineage Protects against Inflammation-Mediated Kidney Injury in Mice

Sakai, Tatsuo, Young, Morag J., Nagase, Miki, Kurihara, Hidetake, Aiba, Atsu

2016

Overview

Macrophage-mediated inflammation has been implicated in various kidney diseases. We previously reported that Rac1, a Rho family small GTP-binding protein, was overactivated in several chronic kidney disease models, and that Rac1 inhibitors ameliorated renal injury, in part via inhibition of inflammation, but the detailed mechanisms have not been clarified. In the present study, we examined whether Rac1 in macrophages effects cytokine production and the inflammatory mechanisms contributing to kidney derangement. Myeloid-selective Rac1 flox control (M-Rac1 FC) and knockout (M-Rac1 KO) mice were generated using the cre-loxP system. Renal function under basal conditions did not differ between M-Rac1 FC and KO mice. Accordingly, lipopolysaccharide (LPS)-evoked kidney injury model was created. LPS elevated blood urea nitrogen and serum creatinine, enhanced expressions of kidney injury biomarkers, Kim-1 and Ngal, and promoted tubular injury in M-Rac1 FC mice. By contrast, deletion of myeloid Rac1 almost completely prevented the LPS-mediated renal impairment. LPS triggered a marked induction of macrophage-derived inflammatory cytokines, IL-6 and TNFα, in M-Rac1 FC mice, which was accompanied by Rac1 activation, stimulation of reduced nicotinamide-adenine dinucleotide phosphate (NADPH) oxidase, and reactive oxygen species overproduction. These changes were inhibited in M-Rac1 KO mice. LPS evoked F4/80-positive macrophages accumulation in the kidney, which was not affected by myeloid Rac1 deficiency. We further tested the role of Rac1 signaling in cytokine production using macrophage cell line, RAW264.7. Exposure to LPS increased IL-6 and TNFα mRNA expression. The LPS-driven cytokine induction was dose-dependently blocked by the Rac1 inhibitor EHT1864, NADPH oxidase inhibitor diphenyleneiodonium, and NF-κB inhibitor BAY11-7082. In conclusion, genetic ablation of Rac1 in the myeloid lineage protected against LPS-induced renal inflammation and injury, by suppressing macrophage-derived cytokines, IL-6 and TNFα, without blocking recruitment. Our data suggest that Rac1 in macrophage is a novel target for the treatment of kidney disease through inhibition of cytokine production.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Analysis / Animal models / Animals / Biology and Life Sciences / Biomarkers / Biomarkers - metabolism / Cell Line / Cell Lineage / Chemokine CCL2 - genetics / Chemokine CXCL1 - genetics / Chemokine CXCL2 - genetics / Chemokines / Chronic kidney failure / Clonal deletion / Creatinine / Culture Media, Conditioned - chemistry / Cytokines / Cytokines - metabolism / Flox / Gene Deletion / Gene expression / Gene Expression Regulation / GTP-binding protein / Inflammation / Inflammation - metabolism / Inflammation - pathology / Inhibition / Inhibitors / Injuries / Interleukin 6 / Interleukin-6 - metabolism / Ischemia / Kidney - pathology / Kidney diseases / Kidneys / Laboratories / Lipopolysaccharides / Macrophages / Macrophages - cytology / Macrophages - metabolism / Male / Medical treatment / Medicine / Medicine and Health Sciences / Mice / Mice, Knockout / Mitogens / Monocytes - cytology / Myeloid Cells - cytology / Myeloid Cells - metabolism / NAD / NAD(P)H oxidase / NADPH Oxidases - metabolism / NADPH-diaphorase / Neuropeptides - genetics / Neuropeptides - metabolism / Neutrophils / NF-κB protein / Nicotinamide / Nitriles - chemistry / Onium Compounds - chemistry / Oxidase / Oxidases / Oxidative stress / Oxygen / Pancreatitis / Phosphates / Physical Sciences / Protein binding / Pyrones - chemistry / Quinolines - chemistry / rac1 GTP-Binding Protein - genetics / rac1 GTP-Binding Protein - metabolism / Rac1 protein / Reactive oxygen species / Reactive Oxygen Species - metabolism / Renal function / Research and Analysis Methods / RNA / Rodents / Signaling / Sulfones - chemistry / Tumor Necrosis Factor-alpha - metabolism / Tumor necrosis factor-α / Urea