title: Pivotal role of serum- and glucocorticoid-inducible kinase 1 in vascular inflammation and atherogenesis.
abstract: OBJECTIVE
  Atherosclerosis, an inflammatory disease of arterial vessel walls, requires migration and matrix metalloproteinase (MMP)-9-dependent invasion of monocytes/macrophages into the vascular wall. MMP-9 expression is stimulated by transcription factor nuclear factor-κB, which is regulated by inhibitor κB (IκB) and thus IκB kinase. Regulators of nuclear factor-κB include serum- and glucocorticoid-inducible kinase 1 (SGK1). The present study explored involvement of SGK1 in vascular inflammation and atherogenesis.
  
  
  RESULTS
  Gene-targeted apolipoprotein E (ApoE)-deficient mice without (apoe(-/-)sgk1(+/+)) or with (apoe(-/-)sgk1(-/-)) additional SGK1 knockout received 16-week cholesterol-rich diet. According to immunohistochemistry atherosclerotic lesions in aorta and carotid artery, vascular CD45(+) leukocyte infiltration, Mac-3(+) macrophage infiltration, vascular smooth muscle cell content, MMP-2, and MMP-9 positive areas in atherosclerotic tissue were significantly less in apoe(-/-)sgk1(-/-)mice than in apoe(-/-)sgk1(+/+)mice. As determined by Boyden chamber, thioglycollate-induced peritonitis and air pouch model, migration of SGK1-deficient CD11b(+)F4/80(+) macrophages was significantly diminished in vitro and in vivo. Zymographic MMP-2 and MMP-9 production, MMP-9 activity and invasion through matrigel in vitro were significantly less in sgk1(-/-) than in sgk1(+/+)macrophages and in control plasmid-transfected or inactive (K127N)SGK1-transfected than in constitutively active (S422D)SGK1-transfected THP-1 cells. Confocal microscopy revealed reduced macrophage number and macrophage MMP-9 content in plaques of apoe(-/-)sgk1(-/-) mice. In THP-1 cells, MMP-inhibitor GM6001 (25 μmol/L) abrogated (S422D)SGK1-induced MMP-9 production and invasion. According to reverse transcription polymerase chain reaction, MMP-9 transcript levels were significantly reduced in sgk1(-/-)macrophages and strongly upregulated in (S422D)SGK1-transfected THP-1 cells compared with control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. According to immunoblotting and confocal microscopy, phosphorylation of IκB kinase and inhibitor κB and nuclear translocation of p50 were significantly lower in sgk1(-/-)macrophages than in sgk1(+/+)macrophages and significantly higher in (S422D)SGK1-transfected THP-1 cells than in control plasmid-transfected or (K127N)SGK1-transfected THP-1 cells. Treatment of (S422D)SGK1-transfected THP-1 cells with IκB kinase-inhibitor BMS-345541 (10 μmol/L) abolished (S422D)SGK1-induced increase of MMP-9 transcription and gelatinase activity.
  
  
  CONCLUSIONS
  SGK1 plays a pivotal role in vascular inflammation during atherogenesis. SGK1 participates in the regulation of monocyte/macrophage migration and MMP-9 transcription via regulation of nuclear factor-κB.
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Oliver
authors.initials: O
authors.lastname: Borst
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Malte
authors.initials: M
authors.lastname: Schaub
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Britta
authors.initials: B
authors.lastname: Walker
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Evi
authors.initials: E
authors.lastname: Schmid
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Patrick
authors.initials: P
authors.lastname: Münzer
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Jakob
authors.initials: J
authors.lastname: Voelkl
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Ioana
authors.initials: I
authors.lastname: Alesutan
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: José M
authors.initials: JM
authors.lastname: Rodríguez
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Sebastian
authors.initials: S
authors.lastname: Vogel
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Tanja
authors.initials: T
authors.lastname: Schoenberger
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Katja
authors.initials: K
authors.lastname: Metzger
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Dominik
authors.initials: D
authors.lastname: Rath
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Anja
authors.initials: A
authors.lastname: Umbach
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Dietmar
authors.initials: D
authors.lastname: Kuhl
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Iris I
authors.initials: II
authors.lastname: Müller
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Peter
authors.initials: P
authors.lastname: Seizer
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Tobias
authors.initials: T
authors.lastname: Geisler
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.).
authors.forename: Meinrad
authors.initials: M
authors.lastname: Gawaz
authors.affiliation: From the Department of Cardiology and Cardiovascular Medicine (O.B., M.S., S.V., T.S., K.M., D.R., I.I.M., P.S., T.G., M.G.), Department of Physiology (O.B., B.W., E.S., P.M., J.V., I.A., A.U., F.L.), Department of Pediatric Surgery and Urology, University Children's Hospital (E.S.), University of Tuebingen, Tuebingen, Germany; Laboratory of Biochemical Genetics and Metabolism, The Rockefeller University of New York (J.M.R.); and Center for Molecular Neurobiology (ZMNH), Institute for Molecular and Cellular Cognition (IMCC), University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany (D.K.). florian.lang@uni-tuebingen.de meinrad.gawaz@med.uni-tuebingen.de.
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D001011:Aorta / Q000201:enzymology / Q000473:pathology; D001018:Aortic Diseases / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D002339:Carotid Arteries / Q000201:enzymology / Q000473:pathology; D002340:Carotid Artery Diseases / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D002460:Cell Line; D002633:Chemotaxis; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D020672:I-kappa B Proteins / Q000378:metabolism; D017874:Immediate-Early Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D007249:Inflammation / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D008264:Macrophages / Q000201:enzymology / Q000473:pathology; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D052002:NF-kappa B p50 Subunit / Q000378:metabolism; D010538:Peritonitis / Q000139:chemically induced / Q000201:enzymology / Q000235:genetics; D058226:Plaque, Atherosclerotic; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013864:Thioglycolates; D014158:Transcription, Genetic; D014162:Transfection; D066253:Vascular Remodeling; D000099184:Serum-Glucocorticoid Regulated Kinases
chemical_list: D001057:Apolipoproteins E; D051550:I-kappa B Kinase; D020672:I-kappa B Proteins; D017874:Immediate-Early Proteins; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D052002:NF-kappa B p50 Subunit; D017346:Protein Serine-Threonine Kinases; D013864:Thioglycolates; D000099184:Serum-Glucocorticoid Regulated Kinases; C017487:2-mercaptoacetate; C522361:MMP2 protein, human; C522362:Mmp2 protein, mouse; C579270:MMP9 protein, human; C501830:Mmp9 protein, mouse
keywords: atherosclerosis; inflammation; migration and invasion protein; serum-glucocorticoid regulated kinase