Korean Journal of Nephrology 2000;19(6):999-1011.

복막 중피세포에서 Nitric Oxide가 VCAM-1의 발현에 미치는 영향 : cGMP 및 NF-kB의 역할 (Nitric Oxide Inhibits VCAM-1 Expression in Human Peritoneal Mesothelial Cells : Possible Role of Cyclic GMP and NF-kB)

김지훈(Jee Hoon Kim),양원석(Won Suk Yang),김순배(Soon Bae Kim),박수길(Soo Gil Park),이상구(Sang Goo Lee),박정식(Jung Sik Park)

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-kB. Cells were exposed to tumor necrosis factor-α (TNF-α) 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 flo#w cytometry. To detect NF-kB binding activity, electrophoretic mobility shift assay(EMSA) was performed. To determine the role of guanylate cyclase or cGMP, inhibitor of guanylate cyclase, 1H-[l, 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-α-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-1β) or lipopolysaccharide(LPS) as well. By EMSA, SIN-1 inhibited the TNF-α-induced NF-kB activity. The 8-bromo-cGMP had no significant effect on TNF-α-induced VCAM-1 mRNA expression and ODQ also had no significant influence on the inhibitory effect of SIN-l. 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-kB activity through cGMP-independent pathway.