title: Tranilast inhibits cytokine-induced nuclear factor kappaB activation in vascular endothelial cells.
abstract: Tranilast [N-(3,4-dimethoxycinnamoyl)anthranilic acid] inhibits vascular inflammation. However, the relevant anti-inflammatory mechanisms are not completely understood. We studied the effects of tranilast on nuclear factor-kappaB (NF-kappaB)-dependent endothelial cell adhesion molecule expression and transcriptional regulation. Cultured human umbilical vein endothelial cells were preincubated with 12.5 to 100 microg/ml tranilast. Tumor necrosis factor-alpha (TNF-alpha)-induced endothelial VCAM-1, ICAM-1, and E-selectin surface expression was inhibited dose dependently. Maximal inhibition achieved with 100 microg/ml tranilast was 38 +/- 6.9, 31.8 +/- 1.5, and 31.9 +/- 1.9%, respectively (mean +/- S.E.M., p < 0.001, n = 5). Secretion of interleukin 6, which is also NF-kappaB-sensitive, was significantly inhibited by tranilast. Endothelial MHC-I expression, which is independent of NF-kappaB, was not inhibited. Although cytokine-induced degradation of NF-kappaB inhibitor proteins (IkappaB-alpha, -beta, and -epsilon), nuclear translocation of NF-kappaB, and binding of NF-kappaB to kappaB cis-acting elements in the adhesion molecule promoters were not affected by tranilast, ICAM-1-kappaB and E-selectin-kappaB reporter gene activity was inhibited by 53% (n = 5, p < 0.01) and 51% (n = 5, p < 0.001), respectively. In contrast, using SP-1 and C/EBP constructs, reporter gene activity was not altered. Expression of the transcriptional coactivator cAMP response element binding protein binding protein (CBP) was inhibited by tranilast, resulting in a loss of interaction between NF-kappaB and CBP. Therefore, in therapeutically relevant concentrations (50 microg/ml), tranilast inhibits NF-kappaB-dependent transcriptional activation by interfering with the NF-kappaB/CBP association. We propose that inhibition of NF-kappaB dependent gene transcription contributes to the anti-inflammatory effects of tranilast.
authors.affiliation: Department of Medicine II, University of Bochum, Bochum, Germany. martin.spieker@ruhr.uni-bochum.de
authors.forename: Martin
authors.initials: M
authors.lastname: Spiecker
authors.forename: Ioana
authors.initials: I
authors.lastname: Lorenz
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Marx
authors.forename: Harald
authors.initials: H
authors.lastname: Darius
mesh_terms: D051697:Activating Transcription Factor 1; D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004268:DNA-Binding Proteins; D019281:Dimerization; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D008025:Ligases / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
chemical_list: D051697:Activating Transcription Factor 1; D015815:Cell Adhesion Molecules; D016207:Cytokines; D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D016328:NF-kappa B; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D062367:ortho-Aminobenzoates; D004247:DNA; D008025:Ligases; C028812:guanosine 3',5'-polyphosphate synthetases; C012293:tranilast