Endothelin 1 transcription is controlled by nuclear factor-kappaB in AGE-stimulated cultured endothelial cells.
Incubation of bovine aortic endothelial cells (BAECs) with erythrocytes from patients with type 2 diabetes induced an increase in endothelin 1 (ET-1) production. The effect of erythrocytes on ET-1 synthesis was dependent on glycemic control. ET-1 levels after incubation with erythrocytes derived from patients with HbA(1c) levels <6% were just half the levels observed after incubation with erythrocytes from patients with HbA(1c) levels >8%. Nepsilon-(carboxymethyl)lysine (CML)-containing protein isolated from patients' erythrocytes induced ET-1, and CML-containing protein-dependent ET-1 induction was blocked by the recombinant decoy peptide soluble receptor for advanced glycation end products (AGEs), which comprises the NH2-terminal Ig domain of the receptor for AGEs. In vitro-generated AGEs induced ET-1 mRNA transcription (nuclear run-on assay and Northern blot) in a time- and dose-dependent manner. Transient transfection of BAECs with a chimeric construct containing the 5' promoter region of the ET-1 gene linked to a reporter gene confirmed that AGE induced ET-1 promoter activity. Electrophoretic mobility shift assay confirmed AGE-inducible binding of members of the nuclear factor-kappab (NF-kappaB) family to a potential binding site at -2,090 bp. Binding was functionally significant because overexpression of the cytoplasmic inhibitor of NF-kappaB or deletion of the NF-kappaB binding site reduced ET-1 induction, whereas overexpression of NF-kappaB p65 induced ET-1 even in the absence of AGEs. Thus, ET-1 transcription is controlled by the AGE-inducible redox-sensitive transcription factor NF-kappaB.
Quehenberger
P
P
Department of Medical and Chemical Laboratory Diagnostics, University of Vienna, Austria.
Bierhaus
A
A
Fasching
P
P
Muellner
C
C
Klevesath
M
M
Hong
M
M
Stier
G
G
Sattler
M
M
Schleicher
E
E
Speiser
W
W
Nawroth
P P
PP
D000818:Animals; D001011:Aorta; D001665:Binding Sites; D002417:Cattle; D002478:Cells, Cultured; D003924:Diabetes Mellitus, Type 2 / Q000097:blood*; D019332:Endothelin-1 / Q000235:genetics*; D004730:Endothelium, Vascular / Q000502:physiology*; D004912:Erythrocytes / Q000502:physiology*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D017930:Genes, Reporter; D006442:Glycated Hemoglobin / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D016328:NF-kappa B / Q000378:metabolism*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D013873:Thionucleotides; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
D019332:Endothelin-1; D006442:Glycated Hemoglobin; D017127:Glycation End Products, Advanced; D016328:NF-kappa B; D020319:Oligodeoxyribonucleotides, Antisense; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides