Anti-Inflammatory Effect of Heat Shock Protein Induction Is Related to Stabilization of IκBα Through Preventing IκB Kinase Activation in Respiratory Epithelial Cells1

Heat shock protein (HSP) induction confers protection against diverse forms of cellular and tissue injury. However, the mechanism by which HSP exerts cytoprotective effects is unclear. Because HSP induction inhibits genetic expression of pro-inflammatory cytokines, the transcription of which is dependent on NF-κB activation, we explored the relationship between the anti-inflammatory effect of HSP induction and the NF-κB/IκBα pathway. Both HS and sodium arsenite treatment increased HSP70 expression time dependently at mRNA and protein levels. Prior induction of HSP suppressed cytokine-induced IL-8 and TNF-α expression at both mRNA and protein levels. Although HSP induction did not affect total cellular expression of NF-κB, TNF-α-induced increase in NF-κB-DNA binding activity and nuclear translocation of the p65 subunit of NF-κB were inhibited by prior HSP induction, suggesting that activation of NF-κB was blocked. Cytokine-induced IκBα phosphorylation and its degradation were blocked in HSP-induced cells. Immune complex kinase assays demonstrated that TNF-α induced increase in IκB kinase activity was suppressed by prior HSP induction. These results suggest that the anti-inflammatory effect of HSP induction in respiratory epithelial cells is related to stabilization of IκBα, possibly through the prevention of IκB kinase activation, which thereby inhibits activation of NF-κB.

[1]  E. Craig,et al.  Protein Folding In Vivo: Unraveling Complex Pathways , 1997, Cell.

[2]  S. Zuckerman,et al.  Transcriptional inhibition of endotoxin–induced monokine synthesis following heat shock in murine peritoneal macrophages , 1992, Journal of leukocyte biology.

[3]  Zhaodan Cao,et al.  TRAF6 is a signal transducer for interleukin-1 , 1996, Nature.

[4]  W. R. Anderson,et al.  Correlative study of adult respiratory distress syndrome by light, scanning, and transmission electron microscopy. , 1992, Ultrastructural pathology.

[5]  E. Craig,et al.  The heat shock response. , 1985, CRC critical reviews in biochemistry.

[6]  Mike Rothe,et al.  IκB Kinase-β: NF-κB Activation and Complex Formation with IκB Kinase-α and NIK , 1997 .

[7]  Hector R. Wong,et al.  Heat shock activates the I-κBα promoter and increases I-κBα mRNA expression , 1999 .

[8]  J. Frydman,et al.  Chaperones get in touch: the Hip-Hop connection. , 1997, Trends in biochemical sciences.

[9]  A. Baldwin,et al.  THE NF-κB AND IκB PROTEINS: New Discoveries and Insights , 1996 .

[10]  A S Slutsky,et al.  Induction of heat stress proteins is associated with decreased mortality in an animal model of acute lung injury. , 1993, The American review of respiratory disease.

[11]  Simon C Watkins,et al.  The heat-shock response attenuates lipopolysaccharide-mediated apoptosis in cultured sheep pulmonary artery endothelial cells. , 1996, American journal of respiratory cell and molecular biology.

[12]  M. Karin,et al.  Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases. , 1997, The New England journal of medicine.

[13]  H. Wong,et al.  The heat shock response inhibits RANTES gene expression in cultured human lung epithelium. , 1998, Journal of immunology.

[14]  Tom Maniatis,et al.  The ubiquitinproteasome pathway is required for processing the NF-κB1 precursor protein and the activation of NF-κB , 1994, Cell.

[15]  T. Maniatis,et al.  The proteasome pathway is required for cytokine-induced endothelial-leukocyte adhesion molecule expression. , 1995, Immunity.

[16]  T. Raffin,et al.  Tumor necrosis factor causes increased pulmonary permeability and edema. Comparison to septic acute lung injury. , 1988, The American review of respiratory disease.

[17]  E. Weibel,et al.  Ultrastructural aspects of pulmonary edema. , 1988, Journal of thoracic imaging.

[18]  Arthur S Slutsky,et al.  Heat stress increases survival rates in lipopolysaccharide-stimulated rats. , 1997, Critical care medicine.

[19]  T. Maniatis,et al.  Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway. , 1995, Genes & development.

[20]  A S Slutsky,et al.  Induction of the heat shock response reduces mortality rate and organ damage in a sepsis‐induced acute lung injury model , 1994, Critical care medicine.

[21]  N. Mackman,et al.  RelB Modulation of IκBα Stability as a Mechanism of Transcription Suppression of Interleukin-1α (IL-1α), IL-1β, and Tumor Necrosis Factor Alpha in Fibroblasts , 1999, Molecular and Cellular Biology.

[22]  Hector R. Wong,et al.  The Heat Shock Response Inhibits Inducible Nitric Oxide Synthase Gene Expression by Blocking Iκ-B Degradation and NF-κB Nuclear Translocation , 1997 .

[23]  H. Wong,et al.  Heat shock activates the I-kappaBalpha promoter and increases I-kappaBalpha mRNA expression. , 1999, Cell stress & chaperones.

[24]  E. Zandi,et al.  The IκB Kinase Complex (IKK) Contains Two Kinase Subunits, IKKα and IKKβ, Necessary for IκB Phosphorylation and NF-κB Activation , 1997, Cell.

[25]  P. Silver.  How proteins enter the nucleus , 1991, Cell.

[26]  Matthias Mann,et al.  IKK-1 and IKK-2: Cytokine-Activated IκB Kinases Essential for NF-κB Activation , 1997 .

[27]  D. Goeddel,et al.  TNF-dependent recruitment of the protein kinase RIP to the TNF receptor-1 signaling complex. , 1996, Immunity.

[28]  J. Wispé,et al.  HEAT SHOCK PROTEIN INDUCTION PROTECTS HUMAN RESPIRATORY EPITHELIUM AGAINST NITRIC OXIDE‐MEDIATED CYTOTOXICITY , 1997, Shock.

[29]  D. Goeddel,et al.  Identification and Characterization of an IκB Kinase , 1997, Cell.

[30]  T. Ikejima,et al.  Interleukin 1 induces a shock-like state in rabbits. Synergism with tumor necrosis factor and the effect of cyclooxygenase inhibition. , 1988, The Journal of clinical investigation.

[31]  D. Gius,et al.  Heat Shock Inhibits Radiation-induced Activation of NF-κB via Inhibition of I-κB Kinase* , 1999, The Journal of Biological Chemistry.

[32]  Hong-Bing Shu,et al.  TRADD–TRAF2 and TRADD–FADD Interactions Define Two Distinct TNF Receptor 1 Signal Transduction Pathways , 1996, Cell.

[33]  C. Dang,et al.  Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. , 1989, The Journal of biological chemistry.

[34]  David M. Rothwarf,et al.  A cytokine-responsive IκB kinase that activates the transcription factor NF-κB , 1997, Nature.

[35]  F. Hartl.  Molecular chaperones in cellular protein folding , 1996, Nature.

[36]  Z. Cao,et al.  IRAK: A Kinase Associated with the Interleukin-1 Receptor , 1996, Science.

[37]  H. Wong,et al.  Stress response decreases NF-kappaB nuclear translocation and increases I-kappaBalpha expression in A549 cells. , 1997, The Journal of clinical investigation.

[38]  J. Pittet,et al.  Biological markers of acute lung injury: prognostic and pathogenetic significance. , 1997, American journal of respiratory and critical care medicine.

[39]  M J May,et al.  NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. , 1998, Annual review of immunology.

[40]  F. Ognibene,et al.  Role of imaging and interventional techniques in the diagnosis of respiratory disease in the immunocompromised host. , 1988, Journal of thoracic imaging.

[41]  E. Abdulla,et al.  Down-regulation of IL-1 beta biosynthesis by inducers of the heat-shock response. , 1988, Journal of immunology.