Attenuation of Ozone-Induced Airway Inflammation and Hyper-Responsiveness by c-Jun NH2 Terminal Kinase Inhibitor SP600125

Ozone has potent oxidizing properties, and exposure to ozone causes airway hyper-responsiveness (AHR) and lung inflammation. We determined the importance of c-Jun NH2 terminal kinase (JNK), a member of the mitogen-activated protein kinase pathway, in ozone-induced AHR and inflammation. SP600125 [anthra[1,9-cd] pyrazol-6 (2H)-one], a specific JNK inhibitor (30 mg/kg) or vehicle, was administered by intraperitoneal injection before and after ozone exposure (3 ppm for 3 h). SP600125 significantly reduced total cells, and neutrophils in bronchoalveolar fluid recovered at 20 to 24 h after exposure and inhibited ozone-induced AHR. Ozone exposure induced activation of JNK in the lung as measured by the expression of phosphorylated-c-Jun, an effect abolished by SP600125. Gene-microarray analysis revealed that ozone increased the expression of over 400 genes by more than 2-fold, including interleukin-6 (IL-6), CXCL1 (keratinocyte cytokine), and CCL2 (monocyte chemoattractant protein-1). SP600125 modulated the expression of a subset of 29 ozone-induced genes; IL-6 and CCL2 expression were further increased, whereas the expression of metallothionein 1, hemopexin, and mitogen-activated 3 kinase 6 was decreased in SP600125-treated ozone-exposed mice. Changes in mRNA for IL-6, CXCL1, and CCL2 were confirmed by real-time polymerase chain reaction. Ozone also decreased the expression of over 500 genes, with the most potent effect on angiopoietin-1. SP600125 modulated the expression of 15 of these genes, and in particular, SP600125 reversed ozone-induced decrease in expression of the redox-sensitive transcription factor, hypoxia-induced factor-1α. This study highlights an important role for JNK in response to oxidative stress through modulation of specific inflammatory and redox mediators. Inhibition of JNK with small molecule kinase inhibitors may be a means of reducing ozone-induced inflammation and AHR.

[1]  Kazuhiro Ito,et al.  Kinase inhibitors and airway inflammation. , 2006, European journal of pharmacology.

[2]  Mark E. Eiswerth,et al.  Impacts of ozone on the activities of asthmatics: revisiting the data. , 2005, Journal of environmental management.

[3]  D. Knecht,et al.  Metallothionein mediates leukocyte chemotaxis , 2005, BMC Immunology.

[4]  J. Haddad,et al.  Cytokines and the regulation of hypoxia-inducible factor (HIF)-1alpha. , 2005, International immunopharmacology.

[5]  I. Adcock,et al.  Potential role of c-Jun NH2-terminal kinase in allergic airway inflammation and remodelling: effects of SP600125. , 2005, European journal of pharmacology.

[6]  B. Witzenbichler,et al.  Protective Role of Angiopoietin-1 in Endotoxic Shock , 2005, Circulation.

[7]  S. Shore,et al.  CXCR2 is essential for maximal neutrophil recruitment and methacholine responsiveness after ozone exposure. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[8]  I. Adcock,et al.  Effect of an inhibitor of Jun N‐terminal protein kinase, SP600125, in single allergen challenge in sensitized rats , 2004, Immunology.

[9]  D. Laskin,et al.  Upregulation of phosphoinositide 3-kinase and protein kinase B in alveolar macrophages following ozone inhalation. Role of NF-κB and STAT-1 in ozone-induced nitric oxide production and toxicity , 2002, Molecular and Cellular Biochemistry.

[10]  A. Yoshimura,et al.  Negative regulation of cytokine signaling and inflammatory diseases. , 2003, Current opinion in pharmacology.

[11]  C. Cross,et al.  Ozone-induced disruptions of lung transcriptomes. , 2003, Biochemical and biophysical research communications.

[12]  Donald Metcalf,et al.  SOCS3 negatively regulates IL-6 signaling in vivo , 2003, Nature Immunology.

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

[14]  P. Cohen,et al.  The specificities of protein kinase inhibitors: an update. , 2003, The Biochemical journal.

[15]  G. Johnson,et al.  Mitogen-Activated Protein Kinase Pathways Mediated by ERK, JNK, and p38 Protein Kinases , 2002, Science.

[16]  S. Gupta,et al.  Amelioration of ozone-induced lung injury by anti-tumor necrosis factor-alpha. , 2002, Toxicological sciences : an official journal of the Society of Toxicology.

[17]  S. Shore,et al.  Ozone-induced airway hyperresponsiveness is reduced in immature mice. , 2002, Journal of applied physiology.

[18]  S. Sur,et al.  CCL7 and CXCL10 Orchestrate Oxidative Stress-Induced Neutrophilic Lung Inflammation1 , 2002, The Journal of Immunology.

[19]  J. Tavernier,et al.  A Mediator Role For Metallothionein in Tumor Necrosis Factor–induced Lethal Shock , 2001, The Journal of experimental medicine.

[20]  David W. Anderson,et al.  SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[21]  S. Shore,et al.  Tumor necrosis factor receptor 2 contributes to ozone-induced airway hyperresponsiveness in mice. , 2001, American journal of respiratory and critical care medicine.

[22]  Hye-Youn Cho,et al.  Ozone-induced lung inflammation and hyperreactivity are mediated via tumor necrosis factor-alpha receptors. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[23]  R. Davis,et al.  Signal Transduction by the JNK Group of MAP Kinases , 2000, Cell.

[24]  J. Gamble,et al.  Angiopoietin-1 Is an Antipermeability and Anti-Inflammatory Agent In Vitro and Targets Cell Junctions , 2000, Circulation research.

[25]  D. Gupta.  LUNG INJURY, INFLAMMATION, AND INFLAMMATORY STIMULI IN RATS EXPOSED TO OZONE , 2000, Journal of toxicology and environmental health. Part A.

[26]  K. Kuwano,et al.  Nitric oxide synthase inhibitors attenuate ozone-induced airway inflammation in guinea pigs. Possible role of interleukin-8. , 2000, American journal of respiratory and critical care medicine.

[27]  Y. Nasuhara,et al.  Inhibition of ozone-induced lung neutrophilia and nuclear factor-kappaB binding activity by vitamin A in rat. , 1999, European journal of pharmacology.

[28]  P. Paggiaro,et al.  Airway inflammatory response to ozone in subjects with different asthma severity. , 1999, The European respiratory journal.

[29]  B. Stripp,et al.  Inflammatory and antioxidant gene expression in C57BL/6J mice after lethal and sublethal ozone exposures. , 1999, Experimental lung research.

[30]  T. Tan,et al.  MAPKKK6, a novel mitogen-activated protein kinase kinase kinase, that associates with MAPKKK5. , 1998, Biochemical and biophysical research communications.

[31]  Jian Wang,et al.  Aldosterone responsiveness of A6 cells is restored by cloned rat mineralocorticoid receptor. , 1998, American journal of physiology. Cell physiology.

[32]  K. Driscoll,et al.  Chemokine regulation of ozone-induced neutrophil and monocyte inflammation. , 1998, American journal of physiology. Lung cellular and molecular physiology.

[33]  C. Scannell,et al.  Effects of ozone on normal and potentially sensitive human subjects. Part I: Airway inflammation and responsiveness to ozone in normal and asthmatic subjects. , 1997, Research report.

[34]  J. Vilček,et al.  TSG-6: an IL-1/TNF-inducible protein with anti-inflammatory activity. , 1997, Cytokine & growth factor reviews.

[35]  I. Adcock,et al.  Ozone induction of cytokine‐induced neutrophil chemoattractant (CINC) and nuclear factor‐κb in rat lung: inhibition by corticosteroids , 1996, FEBS letters.

[36]  I. Adcock,et al.  Dexamethasone inhibits ozone‐induced gene expression of macrophage inflammatory protein‐2 in rat lung , 1995, FEBS letters.

[37]  S. Kleeberger,et al.  Differential susceptibility to ozone-induced airways hyperreactivity in inbred strains of mice. , 1995, Experimental lung research.

[38]  M. Karin,et al.  The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. , 1991, Biochimica et biophysica acta.

[39]  D. Brenner,et al.  Prolonged activation of jun and collagenase genes by tumour necrosis factor-α , 1989, Nature.

[40]  H. Boushey,et al.  O3-induced change in bronchial reactivity to methacholine and airway inflammation in humans. , 1986, Journal of applied physiology.

[41]  P. O'Byrne,et al.  Airway hyperresponsiveness and changes in cell counts in bronchoalveolar lavage after ozone exposure in dogs. , 1984, The American review of respiratory disease.