Korean J Nephrol.  2000 Nov;19(6):999-1011.

Nitric Oxide Inhibits VCAM-1 Expression in Human Peritoneal Mesothelial Cells: Possible Role of Cyclic GMP and NF-kappa B

Affiliations
  • 1Department of Internal Medicine, College of Medicine, University of Ulsan, Seoul, Korea.

Abstract

Leukocyte adhesion to mesothelium is an important step during peritonitis, which may be mediated by adhesion molecules including vascular cell adhesion molecule-1(VCAM-1). Nitric oxide(NO) is known to be an endogenous inhibitor of leukocyte adhesion. We investigated the effect of NO on VCAM-1 expression in cultured human peritoneal mesothelial cells and the possible role of cGMP and NF-kappa B. Cells were exposed to tumor necrosis factor-alpha(TNF-alpha) for the indicated periods in the presence or absence of NO donors, 3-morpholino-sydnonimine (SIN-1) or nitroprusside(NP). The expression of VCAM-1 mRNA and cell surface VCAM-1 molecule was measured by Northern blot analysis and flow cytometry. To detect NF-kappa B binding activity, electrophoretic mobility shift assay(EMSA) was performed. To determine the role of guanylate cyclase or cGMP, inhibitor of guanylate cyclase, 1H-[1, 2, 4] oxadiazolo [4, 3-a] quinoxalin-1-one(ODQ) or analogue of cGMP, 8-bromo-cGMP was used. Both SIN-1 and NP inhibited the TNF-alpha-induced VCAM-1 mRNA expression in a dose dependent manner. SIN-1 also inhibited the expression of cell surface VCAM-1 molecule. Furthermore, SIN-1 and NP inhibited the expression of VCAM-1 mRNA induced by interleukin-1(IL-1beta) or lipopolysaccharide(LPS) as well. By EMSA, SIN-1 inhibited the TNF-alpha-induced NF-kappa B activity. The 8-bromo-cGMP had no significant effect on TNF-alpha-induced VCAM-1 mRNA expression and ODQ also had no significant influence on the inhibitory effect of SIN-1. In conclusion, NO may play an important role in mediating the inflammatory process during peritonitis by down-regulating the mesothelial VCAM-1 expression via suppression of NF-kappa B activity through cGMP- independent pathway.

Keyword

Nitric oxide; Human peritoneal mesothelial cell; VCAM-1

MeSH Terms

Blotting, Northern
Cell Adhesion
Cyclic GMP*
Epithelium
Flow Cytometry
Guanylate Cyclase
Humans*
Leukocytes
Necrosis
Negotiating
NF-kappa B*
Nitric Oxide*
Peritonitis
RNA, Messenger
Tissue Donors
Vascular Cell Adhesion Molecule-1*
Cyclic GMP
Guanylate Cyclase
NF-kappa B
Nitric Oxide
RNA, Messenger
Vascular Cell Adhesion Molecule-1
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