Diabetes-Associated Sustained Activation of the Transcription Factor Nuclear Factor-κB
- Angelika Bierhaus12,
- Stephan Schiekofer12,
- Markus Schwaninger1,
- Martin Andrassy12,
- Per M. Humpert2,
- Jiang Chen12,
- Mei Hong2,
- Thomas Luther3,
- Thomas Henle3,
- Ingrid Klöting4,
- Michael Morcos1,
- Marion Hofmann5,
- Hans Tritschler6,
- Bernd Weigle3,
- Michael Kasper3,
- Mark Smith7,
- George Perry7,
- Ann-Marie Schmidt5,
- David M. Stern5,
- Hans-Ulrich Häring2,
- Erwin Schleicher2 and
- Peter P. Nawroth12
- 1Department of Medicine I and Department of Neurology, University of Heidelberg, Heidelberg, Germany
- 2Department of Medicine IV, University Tübingen, Tübingen, Germany
- 3Department of Anatomy, Department of Immunology, Department of Pathology and Institute of Food Chemistry, Technical University Dresden, Dresden, Germany
- 4Department of Laboratory Animal Science, Institute of Pathophysiology, University Greifswald, Karlsburg, Germany
- 5Columbia University, Department of Physiology, New York, New York
- 6ASTA-Medica, Frankfurt am Main, Germany
- 7Institute of Pathology, Case Western Reserve University, Cleveland, Ohio
Abstract
Activation of the transcription factor nuclear factor-κB (NF-κB) has been suggested to participate in chronic disorders, such as diabetes and its complications. In contrast to the short and transient activation of NF-κB in vitro, we observed a long-lasting sustained activation of NF-κB in the absence of decreased IκBα in mononuclear cells from patients with type 1 diabetes. This was associated with increased transcription of NF-κBp65. A comparable increase in NF-κBp65 antigen and mRNA was also observed in vascular endothelial cells of diabetic rats. As a mechanism, we propose that binding of ligands such as advanced glycosylation end products (AGEs), members of the S100 family, or amyloid-β peptide (Aβ) to the transmembrane receptor for AGE (RAGE) results in protein synthesis–dependent sustained activation of NF-κB both in vitro and in vivo. Infusion of AGE-albumin into mice bearing a β-globin reporter transgene under control of NF-κB also resulted in prolonged expression of the reporter transgene. In vitro studies showed that RAGE-expressing cells induced sustained translocation of NF-κB (p50/p65) from the cytoplasm into the nucleus for >1 week. Sustained NF-κB activation by ligands of RAGE was mediated by initial degradation of IκB proteins followed by new synthesis of NF-κBp65 mRNA and protein in the presence of newly synthesized IκBα and IκBβ. These data demonstrate that ligands of RAGE can induce sustained activation of NF-κB as a result of increased levels of de novo synthesized NF-κBp65 overriding endogenous negative feedback mechanisms and thus might contribute to the persistent NF-κB activation observed in hyperglycemia and possibly other chronic diseases.
Footnotes
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Address correspondence and reprint requests to Peter P. Nawroth, MD, Department of Medicine I, University of Heidelberg, Bergheimer Strasse 58, 69115 Heidelberg, Germany. E-mail: peter_nawroth{at}med.uni-heidelberg.de.
Received for publication 19 October 2000 and accepted in revised form 12 September 2001.
M.S. and G. P. serve as consultants/collaborators for Panacea Pharmaceuticals, Prion Development Laboratories, and Voyager Laboratories. A.-M.S. and D.M.S. are consultants for and have received a research grant from TransTech Pharma. P.P.N. has received a research grant from ASTA-Medica.
Aβ, amyloid-β peptide; AGE, advanced glycosylation end product; BAEC, bovine aortic endothelial cell; β-Gal, β-galactosidase; CML, carboxymethyllysine; DMEM, Dulbecco’s modified Eagle’s medium; ECL, enhanced chemiluminescence; EMSA, electrophoretic mobility shift assay; ERK, extracellular signal-regulated kinases; FCS, fetal calf serum; HO-1, heme oxygenase-1; HPLC, high-performance liquid chromatography; HPRT, hypoxanthine guanine phosphoribosyl transferase; LPS, lipopolysaccharide; NF-κB, nuclear factor-κB; ODN, oligodeoxynucleotide; PBMC, peripheral blood mononuclear cell; PBS, phosphate-buffered saline; PCR, polymerase chain reaction; PMSF, phenylmethylsulfonyl fluoride; [PS]ODN, phosphorothioate oligodeoxynucleotide; RAGE, receptor for AGE; ROS, reactive oxygen species; RT, reverse transcription; rt, room temperature; SMC, smooth muscle cell; sRAGE, soluble RAGE; TBS, Tris-buffered saline; TNF-α, tumor necrosis factor-α.
