Superoxide, H2O2, and iron are required for TNF-alpha-induced MCP-1 gene expression in endothelial cells: role of Rac1 and NADPH oxidase.

Reactive oxygen species (ROS) play an important but not yet fully defined role in the expression of inflammatory genes such as monocyte chemoattractant protein (MCP)-1. We used complementary molecular and biochemical approaches to explore the roles of specific ROS and their molecular linkage to inflammatory signaling in endothelial cells. Adenovirus-mediated expression of superoxide dismutase and catalase inhibited TNF-alpha-induced MCP-1 gene expression, suggesting important roles of superoxide (O(2)(-).) and H(2)O(2) in MCP-1 gene activation. In addition, the iron chelator 1,2-dimethyl-3-hydroxypyridin-4-one and the hydroxyl radical scavengers dimethylthiourea and dimethyl sulfoxide inhibited TNF-alpha-induced MCP-1 expression, suggesting important roles of iron and hydroxyl radicals in inflammatory signal activation. In contrast, scavenging of peroxynitrite with 5,10,15,20-tetrakis-(4-sulfonatophenyl)prophyrinato iron (III) chloride had no effect on TNF-alpha-induced MCP-1 expression. Inhibition of NADPH oxidase, the major oxidase responsible for O(2)(-). generation, with diphenylene iodonium suppressed TNF-alpha-induced MCP-1 mRNA accumulation. Rac1 is an upstream signaling molecule for the activation of NADPH oxidase and O(2)(-). generation. Expression of dominant negative N17Rac1 by adenovirus suppressed TNF-alpha-induced MCP-1 mRNA levels and MCP-1 protein secretion. Expression of N17Rac1 inhibited TNF-alpha-induced MCP-1 and NF-kappaB transcriptional activity. These data suggest that ROS such as superoxide and H(2)O(2) derived from Rac1-activated NADPH oxidase mediate TNF-alpha-induced MCP-1 expression in endothelial cells.

[1]  Qiang Zhang,et al.  Rac1 and Superoxide Are Required for the Expression of Cell Adhesion Molecules Induced by Tumor Necrosis Factor-α in Endothelial Cells , 2003, Journal of Pharmacology and Experimental Therapeutics.

[2]  C. Kunsch,et al.  Laminar Flow Induction of Antioxidant Response Element-mediated Genes in Endothelial Cells , 2003, The Journal of Biological Chemistry.

[3]  H. Kovacic,et al.  Rac-Dependent Monocyte Chemoattractant Protein-1 Production Is Induced by Nutrient Deprivation , 2002, Circulation research.

[4]  A. Shah,et al.  Essential Role of the NADPH Oxidase Subunit p47phox in Endothelial Cell Superoxide Production in Response to Phorbol Ester and Tumor Necrosis Factor-&agr; , 2002, Circulation research.

[5]  S. Davidge,et al.  Peroxynitrite increases iNOS through NF-kappaB and decreases prostacyclin synthase in endothelial cells. , 2002, American journal of physiology. Cell physiology.

[6]  M. Galiñanes,et al.  Peroxynitrite Is an Essential Component of Cytokines Production Mechanism in Human Monocytes through Modulation of Nuclear Factor-κB DNA Binding Activity* , 2002, The Journal of Biological Chemistry.

[7]  W. Aird,et al.  Vascular endothelial growth factor induces manganese‐superoxide dismutase expression in endothelial cells by a Racl‐regulated NADPH oxidase‐dependent mechanism , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  J. Filep,et al.  Peroxynitrite mediates cytokine‐induced IL‐8 gene expression and production by human leukocytes , 2001, Journal of leukocyte biology.

[9]  K. Irani,et al.  Rac1 Regulates Stress-induced, Redox-dependent Heat Shock Factor Activation* , 2000, The Journal of Biological Chemistry.

[10]  Jang-Soo Chun,et al.  Tumor Necrosis Factor-α Generates Reactive Oxygen Species via a Cytosolic Phospholipase A2-linked Cascade* , 2000, The Journal of Biological Chemistry.

[11]  K. Irani,et al.  Rac1 inhibits TNF‐α‐induced endothelial cell apoptosis: dual regulation by reactive oxygen species , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[12]  R. Medford,et al.  NF- kappa B independent suppression of endothelial vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 gene expression by inhibition of flavin binding proteins and superoxide production. , 2000, Journal of molecular and cellular cardiology.

[13]  W. Kox,et al.  Secretion of MCP-1 and IL-6 by cytokine stimulated production of reactive oxygen species in endothelial cells , 2000, Molecular and Cellular Biochemistry.

[14]  B. Rollins,et al.  Monocyte Chemoattractant Protein 1–Dependent Leukocytic Infiltrates Are Responsible for Autoimmune Disease in Mrl-Faslpr Mice , 1999, The Journal of experimental medicine.

[15]  I. Fridovich,et al.  Superoxide and Iron: Partners in Crime , 1999, IUBMB life.

[16]  K. Irani,et al.  Shear-induced tyrosine phosphorylation in endothelial cells requires Rac1-dependent production of ROS. , 1999, American journal of physiology. Cell physiology.

[17]  B. Rollins,et al.  MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B. , 1999, The Journal of clinical investigation.

[18]  R. Eisenstein,et al.  Iron regulatory proteins, iron responsive elements and iron homeostasis. , 1998, The Journal of nutrition.

[19]  P. Libby,et al.  Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. , 1998, Molecular cell.

[20]  M. Currie,et al.  Characterization of the Cytoprotective Action of Peroxynitrite Decomposition Catalysts* , 1998, The Journal of Biological Chemistry.

[21]  V. Ferrans,et al.  Protection from reoxygenation injury by inhibition of rac1. , 1998, The Journal of clinical investigation.

[22]  B. Rollins,et al.  Abnormalities in Monocyte Recruitment and Cytokine Expression in Monocyte Chemoattractant Protein 1–deficient Mice , 1998, The Journal of experimental medicine.

[23]  R. Medford,et al.  E-selectin gene expression in vascular smooth muscle cells. Evidence for a tissue-specific repressor protein. , 1997, Circulation research.

[24]  V. Ferrans,et al.  rac1 regulates a cytokine-stimulated, redox-dependent pathway necessary for NF-kappaB activation , 1996, Molecular and cellular biology.

[25]  J. Imlay,et al.  Superoxide accelerates DNA damage by elevating free-iron levels. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[26]  V. Ferrans,et al.  Regulation of reactive-oxygen-species generation in fibroblasts by Rac1. , 1996, The Biochemical journal.

[27]  B. Freeman,et al.  Nitric oxide regulation of tissue free radical injury. , 1996, Chemical research in toxicology.

[28]  T. Nagaya,et al.  Antioxidants inhibit tumor necrosis factor-alpha mediated stimulation of interleukin-8, monocyte chemoattractant protein-1, and collagenase expression in cultured human synovial cells. , 1996, The Journal of rheumatology.

[29]  A. Ben-Baruch,et al.  Chemokines: progress toward identifying molecular targets for therapeutic agents. , 1996, Trends in biotechnology.

[30]  J. Westwick,et al.  Regulation and expression of chemokines: potential role in glomerulonephritis , 1996, Journal of leukocyte biology.

[31]  V. Ferrans,et al.  Requirement for Generation of H2O2 for Platelet-Derived Growth Factor Signal Transduction , 1995, Science.

[32]  Hoffbrand Av,et al.  Iron chelation therapy , 1995, Current opinion in hematology.

[33]  S. Ohno,et al.  NF-kappa B and Sp1 regulate transcription of the human monocyte chemoattractant protein-1 gene. , 1994, Journal of immunology.

[34]  D. Graves,et al.  Expression of monocyte chemoattractant protein-1 in delayed type hypersensitivity reactions in the skin. , 1994, Laboratory investigation; a journal of technical methods and pathology.

[35]  N. Sepp,et al.  Characterization of expression and modulation of cell adhesion molecules on an immortalized human dermal microvascular endothelial cell line (HMEC-1). , 1994, The Journal of investigative dermatology.

[36]  P. Kaminski,et al.  NADH oxidoreductase is a major source of superoxide anion in bovine coronary artery endothelium. , 1994, The American journal of physiology.

[37]  R. Alexander,et al.  Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. , 1993, The Journal of clinical investigation.

[38]  Yun Xing,et al.  Oxygen radicals as second messengers for expression of the monocyte chemoattractant protein, JE/MCP-1, and the monocyte colony-stimulating factor, CSF-1, in response to tumor necrosis factor-alpha and immunoglobulin G. Evidence for involvement of reduced nicotinamide adenine dinucleotide phosphate ( , 1993, The Journal of clinical investigation.

[39]  R. Terkeltaub,et al.  Production of monocyte chemoattractant protein-1 by inflamed synovial tissue and cultured synoviocytes. , 1992, Journal of immunology.

[40]  A. J. Valente,et al.  Expression of monocyte chemoattractant protein 1 mRNA in human idiopathic pulmonary fibrosis. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[41]  A. Abo,et al.  Activation of the NADPH oxidase involves the small GTP-binding protein p21rac1 , 1991, Nature.

[42]  S. Coughlin,et al.  Monocyte chemoattractant protein-1 in human atheromatous plaques. , 1991, The Journal of clinical investigation.

[43]  E. Ruuge,et al.  A simple assay of the superoxide generation rate with Tiron as an EPR-visible radical scavenger. , 1986, Biochemistry international.

[44]  R. Medford,et al.  Rac1 and superoxide are required for the expression of cell adhesion molecules induced by tumor necrosis factor-alpha in endothelial cells. , 2003, The Journal of pharmacology and experimental therapeutics.

[45]  J. Korn,et al.  Induction of heme oxygenase-1 by hypoxia and free radicals in human dermal fibroblasts. , 2000, American journal of physiology. Cell physiology.

[46]  K. Irani,et al.  The actin cytoskeleton reorganization induced by Rac1 requires the production of superoxide. , 1999, Antioxidants & redox signaling.

[47]  B. Lucchesi.  Free radicals and tissue injury , 1998 .

[48]  H. Forman,et al.  Oxidants as stimulators of signal transduction. , 1997, Free radical biology & medicine.

[49]  B Dewald,et al.  Human chemokines: an update. , 1997, Annual review of immunology.

[50]  K. Takeshige,et al.  Activation of the phagocyte NADPH oxidase. , 1996, The Journal of toxicological sciences.