title: Deletion of Rac1GTPase in the Myeloid Lineage Protects against Inflammation-Mediated Kidney Injury in Mice.
abstract: 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.
authors.affiliation: Department of Anatomy and Life Structure, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Nagase
authors.affiliation: Department of Anatomy and Life Structure, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Hidetake
authors.initials: H
authors.lastname: Kurihara
authors.affiliation: Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Atsu
authors.initials: A
authors.lastname: Aiba
authors.affiliation: Centre for Endocrinology and Metabolism, Hudson Institute of Medical Research and Monash University, Clayton, Victoria, Australia.
authors.forename: Morag J
authors.initials: MJ
authors.lastname: Young
authors.affiliation: Department of Anatomy and Life Structure, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Sakai
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D019070:Cell Lineage; D018932:Chemokine CCL2 / Q000235:genetics; D054360:Chemokine CXCL1 / Q000235:genetics; D054426:Chemokine CXCL2 / Q000235:genetics; D017077:Culture Media, Conditioned / Q000737:chemistry; D016207:Cytokines / Q000378:metabolism; D017353:Gene Deletion; D005786:Gene Expression Regulation; D007249:Inflammation / Q000378:metabolism / Q000473:pathology*; D015850:Interleukin-6 / Q000378:metabolism*; D007668:Kidney / Q000473:pathology*; D008070:Lipopolysaccharides; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009000:Monocytes / Q000166:cytology; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism; D009479:Neuropeptides / Q000235:genetics* / Q000378:metabolism*; D009570:Nitriles / Q000737:chemistry; D009861:Onium Compounds / Q000737:chemistry; D011753:Pyrones / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism; D013450:Sulfones / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000235:genetics* / Q000378:metabolism*
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D015415:Biomarkers; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D054360:Chemokine CXCL1; D054426:Chemokine CXCL2; D017077:Culture Media, Conditioned; C475895:Cxcl1 protein, mouse; C404915:Cxcl2 protein, mouse; D016207:Cytokines; C506907:EHT 1864; D015850:Interleukin-6; D008070:Lipopolysaccharides; D009479:Neuropeptides; D009570:Nitriles; D009861:Onium Compounds; D011753:Pyrones; D011804:Quinolines; C484602:Rac1 protein, mouse; D017382:Reactive Oxygen Species; D013450:Sulfones; D014409:Tumor Necrosis Factor-alpha; C007517:diphenyleneiodonium; D019255:NADPH Oxidases; D020830:rac1 GTP-Binding Protein