Thrombomodulin has a central role in the regulation of coagulation through its abilities to promote generation of the potent anticoagulant activated protein C. Because little is known about monocyte thrombomodulin expression and its regulatory mechanism by lipopolysaccharide, we investigated the effect of lipopolysaccharide on monocyte's thrombomodulin expression. Lipopolysaccharide reduced the surface thrombomodulin expression of human peripheral blood monocytes in a dose-dependent and time-dependent manner, regardless of the addition of serum. The surface thrombomodulin activity was comparably decreased in monocytes incubated with lipopolysaccharide. Blocking nuclear factor-κB by MG132 or aurine tricarboxylic acid effectively inhibited the lipopolysaccharide-induced surface thrombomodulin down-regulation of monocytes. Lipopolysaccharide inactivation by polymyxin B in the supernatants from the lipopolysaccharide-stimulated cultures still reduced the surface thrombomodulin expression of monocytes, suggesting a role for soluble mediators in the down-regulation of thrombomodulin. The lipopolysaccharide-induced thrombomodulin surface expression and the mRNA levels of the monocytic leukemic cell line (THP-1) were decreased in serum-depleted culture, while they were increased in medium containing 10% serum. We conclude that lipopolysaccharide down-regulates the thrombomodulin expression of monocytes and that nuclear factor-κB is a critical mediator of the repression of thrombomodulin by lipopolysaccharide. Regulation of the THP-1 thrombomodulin expression by lipopolysaccharide depends on the presence of serum.