Effects of antioxidants and NO on TNF- α-induced adhesion molecule expression in human pulmonary microvascular endothelial cells

• Published in: Respiratory medicine 2005-05, Vol.99 (5), p.580-591
• Main Authors: Jiang, Mi-Zu, Tsukahara, Hirokazu, Hayakawa, Kazuyo, Todoroki, Yukiko, Tamura, Satoshi, Ohshima, Yusei, Hiraoka, Masahiro, Mayumi, Mitsufumi
• Format: Article
• Language: English
• Subjects: Antioxidants - pharmacology; Biological and medical sciences; Cell Adhesion Molecules - analysis; Cell Adhesion Molecules - genetics; Cell Nucleus - chemistry; Cytokine; Cytoplasm - chemistry; E-Selectin - analysis; E-Selectin - genetics; Endothelial activation; Endothelial Cells - drug effects; Endothelial Cells - immunology; Endothelium, Vascular - drug effects; Endothelium, Vascular - immunology; Fluorescent Antibody Technique; HL-60 Cells; Human; Humans; Intercellular Adhesion Molecule-1 - analysis; Intercellular Adhesion Molecule-1 - genetics; Medical sciences; NF-kappa B - analysis; Nitric Oxide - pharmacology; Nitric Oxide Donors - pharmacology; Pneumology; Pulmonary inflammation; Reactive oxygen intermediates; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Tumor Necrosis Factor-alpha - immunology; Tumor Necrosis Factor-alpha - pharmacology; Vascular Cell Adhesion Molecule-1 - analysis; Vascular Cell Adhesion Molecule-1 - genetics
• ISSN: 0954-6111; 1532-3064
• DOI: 10.1016/j.rmed.2004.10.007

Pro-inflammatory cytokines initiate the vascular inflammatory response via upregulation of adhesion molecules on the endothelium. Recent observations suggest that reactive oxygen intermediates may play a pivotal role in TNF- α signaling and upregulate gene expression. We therefore evaluated the effects of pyrrolidine dithiocarbamate (PDTC; 0.1 mM) and spermine NONOate (Sper-NO; 1 mM) on adhesion molecule expression and nuclear factor kappa B (NF- κB) activation induced by TNF- α(10 ng/ml) in cultured human pulmonary microvascular endothelial cells (PMVEC). Treatment of cells with TNF- α for 4 h significantly induced the surface expression of E-selectin and ICAM-1. Treatment with TNF- α for 8 h significantly induced the surface expression of E-selectin, ICAM-1 and VCAM-1. The upregulation of these adhesion molecules was suppressed significantly by pretreatment with PDTC or Sper-NO for 1 h. 8-Bromo-cyclic GMP (1 mM) had no such effect, suggesting that the NO donor's effect was non-cGMP-dependent. The mRNA expression of E-selectin, ICAM-1 and VCAM-1, and activation of NF- κB induced by TNF- α for 2 h were decreased significantly by the above two pretreatments. N-acetylcysteine (10 mM) and S-nitroso- N-acetylpenicillamine (1 mM) had little inhibitory effects on the cell surface and mRNA expression of these adhesion molecules stimulated by TNF- α. Treatment with TNF- α for 4 h enhanced HL-60 leukocyte adhesion to human PMVEC, the effect of which was inhibited significantly by pretreatment with PDTC or Sper-NO. These findings indicate that both cell surface and mRNA expression of adhesion molecules in human PMVEC induced by TNF- α are inhibited significantly by pretreatment with PDTC or Sper-NO, possibly in part through blocking the activation of NF- κB. Although our in vitro results cannot be directly extrapolated to the in vivo situation, they suggest a potential therapeutic approach for intervention in cytokine-mediated inflammatory processes in the human lung.