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Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation.

Publication ,  Journal Article
Hall, G; Singh, IS; Hester, L; Hasday, JD; Rogers, TB
Published in: Am J Physiol Heart Circ Physiol
November 2005

TNF-alpha is recognized as a significant contributor to myocardial dysfunction. Although several studies suggest that members of the NF-kappaB family of transcription factors are essential regulators of myocardial TNF-alpha gene expression, recent developments in our understanding of the modulation of NF-kappaB activity through posttranslational modification of NF-kappaB subunits suggest that the present view of NF-kappaB-dependent cytokine expression in heart is incomplete. Therefore, the goal of the present study was to examine the role of p65 subunit phosphorylation in the regulation of TNF-alpha production in cultured neonatal ventricular myocytes. Bacterial LPS-induced TNF-alpha production is accompanied by a 12-fold increase in phosphorylation of p65 at Ser536, a modification associated with enhancement of p65 transactivation potential. Pharmacological inhibition of IKK-beta reduced LPS-induced TNF-alpha production 38-fold, TNF-alpha mRNA levels 6-fold, and IkappaB-alpha phosphorylation 5-fold and degraded IkappaB-alpha 2-fold and p65 phosphorylation 6-fold. Overexpression of dominant-negative p65 reduced TNF-alpha production 3.5-fold, whereas overexpression of dominant-negative IKK-beta reduced LPS-induced TNF-alpha production 2-fold and p65 phosphorylation 2-fold. Overexpression of dominant-negative IKK-alpha had no effect on p65 phosphorylation or TNF-alpha production, revealing that IKK-beta, not IKK-alpha, plays a central role in regulation of p65 phosphorylation at Ser536 and TNF-alpha production in heart. Finally, we demonstrated, using a chromatin immunoprecipitation assay, that LPS stimulates recruitment of Ser536-phosphorylated p65 to the TNF-alpha gene promoter in cardiac myocytes. Taken together, these data provide compelling evidence for the role of NF-kappaB signaling in TNF-alpha gene expression in heart and highlight the importance of this proinflammatory gene-regulatory pathway as a potential therapeutic target in the management of cytokine-induced myocardial dysfunction.

Duke Scholars

Published In

Am J Physiol Heart Circ Physiol

DOI

ISSN

0363-6135

Publication Date

November 2005

Volume

289

Issue

5

Start / End Page

H2103 / H2111

Location

United States

Related Subject Headings

  • Tumor Necrosis Factor-alpha
  • Signal Transduction
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • NF-kappaB-Inducing Kinase
  • Myocytes, Cardiac
  • Myocardium
  • Mutagenesis, Site-Directed
  • Mice
  • Lipopolysaccharides
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Hall, G., Singh, I. S., Hester, L., Hasday, J. D., & Rogers, T. B. (2005). Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation. Am J Physiol Heart Circ Physiol, 289(5), H2103–H2111. https://doi.org/10.1152/ajpheart.00393.2005
Hall, Gentzon, Ishwar S. Singh, Lisa Hester, Jeffery D. Hasday, and Terry B. Rogers. “Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation.Am J Physiol Heart Circ Physiol 289, no. 5 (November 2005): H2103–11. https://doi.org/10.1152/ajpheart.00393.2005.
Hall G, Singh IS, Hester L, Hasday JD, Rogers TB. Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation. Am J Physiol Heart Circ Physiol. 2005 Nov;289(5):H2103–11.
Hall, Gentzon, et al. “Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation.Am J Physiol Heart Circ Physiol, vol. 289, no. 5, Nov. 2005, pp. H2103–11. Pubmed, doi:10.1152/ajpheart.00393.2005.
Hall G, Singh IS, Hester L, Hasday JD, Rogers TB. Inhibitor-kappaB kinase-beta regulates LPS-induced TNF-alpha production in cardiac myocytes through modulation of NF-kappaB p65 subunit phosphorylation. Am J Physiol Heart Circ Physiol. 2005 Nov;289(5):H2103–H2111.

Published In

Am J Physiol Heart Circ Physiol

DOI

ISSN

0363-6135

Publication Date

November 2005

Volume

289

Issue

5

Start / End Page

H2103 / H2111

Location

United States

Related Subject Headings

  • Tumor Necrosis Factor-alpha
  • Signal Transduction
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • NF-kappaB-Inducing Kinase
  • Myocytes, Cardiac
  • Myocardium
  • Mutagenesis, Site-Directed
  • Mice
  • Lipopolysaccharides