Redox regulation of reactive oxygen species-induced p38 MAP kinase activation and barrier dysfunction in lung microvascular endothelial cells.

Reactive oxygen species (ROS)-mediated compromise of endothelial barrier integrity has been implicated in a number of pulmonary disorders, including adult respiratory distress syndrome, pulmonary edema, and vasculitis. The mechanisms by which ROS increase endothelial permeability are unclear. We hypothesized that ROS-induced changes in cellular redox status (thiols) may contribute to endothelial barrier dysfunction. To test this hypothesis, we used N-acetylcysteine (NAC) and diamide to modulate intracellular levels of cellular glutathione (GSH) and investigated hydrogen peroxide (H(2)O(2))-mediated mitogen-activated protein kinase (MAPK) activation and transendothelial electrical resistance (TER). Exposure of bovine lung microvascular endothelial cells (BLMVECs) to H(2)O(2), in a dose- and time-dependent fashion, increased endothelial permeability. Pretreatment of BLMVECs with NAC (5 mM) for 1 h resulted in partial attenuation of H(2)O(2)-induced TER (a measure of increase in permeability) and GSH. Furthermore, treatment of BLMVECs with diamide, which is known to reduce the intracellular GSH, resulted in significant reduction in TER, which was prevented by NAC. To understand further the role of MAPKs in ROS-induced barrier dysfunction, we examined the role of extracellular signal-regulated kinase (ERK) and p38 MAPK on H(2)O(2)- and diamide-mediated permeability changes. Both H(2)O(2) and diamide, in a dose-dependent manner, activated ERK and p38 MAPK in BLMVECs. However, SB203580, an inhibitor of p38 MAPK, but not PD98059, blocked H(2)O(2)- and diamide-induced TER. Also, NAC prevented H(2)O(2)- and diamide-induced p38 MAPK, but not ERK activation. These results suggest a role for redox regulation of p38 MAPK in ROS-dependent endothelial barrier dysfunction.

[1]  R. Flavell,et al.  c-Jun NH2-Terminal Kinase Is Essential for the Regulation of AP-1 by Tumor Necrosis Factor , 2003, Molecular and Cellular Biology.

[2]  J. Travers,et al.  Hyperoxia alters phorbol ester-induced phospholipase D activation in bovine lung microvascular endothelial cells. , 2003, Antioxidants & redox signaling.

[3]  K. Storey,et al.  Mitogen-activated protein kinases: new signaling pathways functioning in cellular responses to environmental stress , 2003, Journal of Experimental Biology.

[4]  M. Ahmad,et al.  Molecular mechanisms of N-acetylcysteine actions , 2003, Cellular and Molecular Life Sciences CMLS.

[5]  H. Forman,et al.  Glutathione in Defense and Signaling , 2002 .

[6]  H. Kamata,et al.  Multiple Redox Regulation of the Cellular Signaling System Linked to AP‐1 and NFκB: Effects of N‐Acetylcysteine and H2O2 on the Receptor Tyrosine Kinases, the MAP Kinase Cascade, and IκB Kinases , 2002, Annals of the New York Academy of Sciences.

[7]  W. J. Esselman,et al.  Inhibition of PTPs by H(2)O(2) regulates the activation of distinct MAPK pathways. , 2002, Free radical biology & medicine.

[8]  F. Liu,et al.  Critical involvement of p38 MAP kinase in pertussis toxin‐induced cytoskeletal reorganization and lung permeability , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[9]  H. Forman,et al.  Glutathione in defense and signaling: lessons from a small thiol. , 2002, Annals of the New York Academy of Sciences.

[10]  P. Vincent,et al.  p38 MAPK activation by TGF-beta1 increases MLC phosphorylation and endothelial monolayer permeability. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[11]  S. Dudek,et al.  Cytoskeletal regulation of pulmonary vascular permeability. , 2001, Journal of applied physiology.

[12]  R. Wadgaonkar,et al.  Role of p38 MAP kinase in diperoxovanadate-induced phospholipase D activation in endothelial cells. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[13]  C M Doerschuk,et al.  The p38 Mitogen-Activated Protein Kinase Mediates Cytoskeletal Remodeling in Pulmonary Microvascular Endothelial Cells Upon Intracellular Adhesion Molecule-1 Ligation1 , 2001, The Journal of Immunology.

[14]  M. Yoshizumi,et al.  Antioxidants inhibit JNK and p38 MAPK activation but not ERK 1/2 activation by angiotensin II in rat aortic smooth muscle cells. , 2001, Hypertension research : official journal of the Japanese Society of Hypertension.

[15]  H. Lum,et al.  Oxidant stress and endothelial cell dysfunction. , 2001, American journal of physiology. Cell physiology.

[16]  E. Levin,et al.  Estrogen Signals to the Preservation of Endothelial Cell Form and Function* , 2000, The Journal of Biological Chemistry.

[17]  B. Fanburg,et al.  Reactive oxygen species in cell signaling. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[18]  M. Wilson,et al.  Constitutive and growth factor-regulated phosphorylation of caveolin-1 occurs at the same site (Tyr-14) in vivo: identification of a c-Src/Cav-1/Grb7 signaling cassette. , 2000, Molecular endocrinology.

[19]  N. Parinandi,et al.  Involvement of c-Src in diperoxovanadate-induced endothelial cell barrier dysfunction. , 2000, American journal of physiology. Lung cellular and molecular physiology.

[20]  P. Brecher,et al.  Distinct Effects of N-Acetylcysteine and Nitric Oxide on Angiotensin II-induced Epidermal Growth Factor Receptor Phosphorylation and Intracellular Ca2+ Levels* , 2000, The Journal of Biological Chemistry.

[21]  M. Yaffe,et al.  MAP kinase pathways activated by stress: the p38 MAPK pathway. , 2000, Critical care medicine.

[22]  Michael Karin,et al.  The Beginning of the End: IκB Kinase (IKK) and NF-κB Activation* , 1999, The Journal of Biological Chemistry.

[23]  K. Roebuck Oxidant stress regulation of IL-8 and ICAM-1 gene expression: differential activation and binding of the transcription factors AP-1 and NF-kappaB (Review). , 1999, International journal of molecular medicine.

[24]  J. Garcia,et al.  Hydrogen peroxide stimulates tyrosine phosphorylation of focal adhesion kinase in vascular endothelial cells. , 1999, American journal of physiology. Lung cellular and molecular physiology.

[25]  K. Aoshiba,et al.  Thiol depletion induces apoptosis in cultured lung fibroblasts. , 1999, American journal of respiratory cell and molecular biology.

[26]  N. Parinandi,et al.  Phospholipase D activation in endothelial cells is redox sensitive. , 1999, Antioxidants & redox signaling.

[27]  K. Roebuck Regulation of interleukin-8 gene expression. , 1999, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[28]  H. Kamata,et al.  Redox regulation of cellular signalling. , 1999, Cellular signalling.

[29]  M. Grisham,et al.  Differential regulation of extracellular signal-regulated kinase and nuclear factor-kappa B signal transduction pathways by hydrogen peroxide and tumor necrosis factor. , 1998, Archives of biochemistry and biophysics.

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

[31]  C. Der,et al.  The Mitogen-activated Protein Kinase Phosphatases PAC1, MKP-1, and MKP-2 Have Unique Substrate Specificities and Reduced Activity in Vivo toward the ERK2 sevenmaker Mutation (*) , 1996, The Journal of Biological Chemistry.

[32]  J. Landry,et al.  Modulation of actin dynamics during stress and physiological stimulation by a signaling pathway involving p38 MAP kinase and heat-shock protein 27. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[33]  A. Bridges,et al.  A synthetic inhibitor of the mitogen-activated protein kinase cascade. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[34]  H. Kawasaki,et al.  Evidence for Multiple Activators for Stress-activated Protein Kinases/c-Jun Amino-terminal Kinases. , 1995, The Journal of Biological Chemistry.

[35]  E. Krebs,et al.  The MAPK signaling cascade , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[36]  Jerry L. Adams,et al.  A protein kinase involved in the regulation of inflammatory cytokine biosynthesis , 1994, Nature.

[37]  C. N. Coleman,et al.  X-irradiation, phorbol esters, and H2O2 stimulate mitogen-activated protein kinase activity in NIH-3T3 cells through the formation of reactive oxygen intermediates. , 1994, Cancer research.

[38]  Ivar Giaever,et al.  A morphological biosensor for mammalian cells , 1993, Nature.

[39]  I. Giaever,et al.  Micromotion of mammalian cells measured electrically. , 1991, Proceedings of the National Academy of Sciences of the United States of America.