Antioxidants inhibit monocyte adhesion by suppressing nuclear factor-kappa B mobilization and induction of vascular cell adhesion molecule-1 in endothelial cells stimulated to generate radicals.

Abstract

Cell adhesion to endothelial cells stimulated by tumor necrosis factor-a (TNF) is due to induction of surface receptors, such as vascular cell adhesion molecule-1 (VCAM-1). The antiaxidant pyrrolidine dithiocarbamate (PDTC) specifically inhibits activation of nuclear factor-KB (NF-KB). Since *B motifs are present in VCAM-1 and intercellular adhesion molecule-1 (ICAM-1) promoters, we used PDTC to study the regulatory mechanisms of VCAM-1 and ICAM-1 induction and subsequent monocyte adhesion in TNF-treated human umbilical vein endothelial cells (HUVECs). PDTC or JV-acetylcysteine dose dependently reduced TNF-induced VCAM-1 but not ICAM-1 surface protein (also in human umbilical arterial endothelial cells) and mRNA expression (by 70% at 100 junol/L PDTC) in HUVECs as assessed by flow cytometry and porymerase chain reaction. Gel-shift analysis in HUVECs demonstrated that PDTC prevented NF-KB mobilization by TNF, suggesting that only VCAM-1 induction was controlled by NF-KB. Since HUVECs released superoxide anions in response to TNF, and H 2 O 2

Differential chemokine receptor expression and function in human monocyte subpopulations

“…cDNA was amplified with Taq polymerase and primers by 36 cycles in a thermocycler 480 (Perkin-Elmer-Cetus) set to 95°C (30 s), 58°C annealing (60 s), 72°C extension (60 s). Primers were synthesized according to sequences with minimal homology for CCR2 [24] with a product size of 557 bp, or were as described for CCR1 and CCR5 [27] or ␤-actin [28]. PCR products were analyzed by 1.25% (CCR2 and ␤-actin) or 2.0% (CCR1 and CCR5) agarose gel electrophoresis, and were quantitated by ultraviolet detection at 260 nm after separation by high-performance liquid chromatography (HPLC) on DEAE columns (Perkin-Elmer), as described in detail [28][29][30].…”

“…Isolation and culture [28] of human umbilical vein endothelial cells (HUVEC) and transendothelial migration assays [24,32] were performed as described.…”

Inhibition of intimal hyperplasia after balloon injury in rat carotid artery model using cis-element ‘decoy’ of nuclear factor-kB binding site as a novel molecular strategy

“…In vascular cells, NF B has been shown to regulate the expression of vascular adhesion molecule-1 (VCAM-1) and intracellular adhesion molecule-1 (ICAM-1) as a part of the inflammatory response. [18][19][20] It is also known that neutralizing antibody to ICAM-1 inhibits intimal hyperplasia after balloon injury of the carotid artery in rats. 21 In this study, to evaluate the therapeutic benefit of inhibition of NF B in neointimal formation after balloon injury, we utilized synthetic double-stranded DNA with high affinity for NF B as a 'decoy' cis element to bind the transcription factor and to block the activation of genes.…”

“…From this viewpoint, we focused on adhesion molecules, as it is known that NFkB regulates the expression of ICAM-1 and VCAM-1, [18][19][20] whose expression is increased in vascular cells after balloon injury. 15,25 Upregulation of these adhesion molecules is considered to be closely implicated in neointimal thickening after arterial injury.…”

α-Lipoic acid attenuates LPS-induced inflammatory responses by activating the phosphoinositide 3-kinase/Akt signaling pathway

“…Intracellular antioxidants may affect cellular redox environment and thus attenuate the response of endothelial cells to oxidative stress and inflammatory cytokines. For example, the antioxidants pyrrolidine dithiocarbamate, N-acetyl-L-cysteine, and ␣-lipoic acid (LA; 1,2-dithiolane-3-pentanoic acid) inhibit cytokine-and LPS-induced CAM expression in various cell types (9)(10)(11) and animal models (12)(13)(14).…”