By study on the effect of
anisodamine on
lipopolysaccharide-induced expression of
tissue factor (TF) in vascular endothelial cells (EC), the mechanism of
anisodamine antithrombosis, as well as in the treatment of bacteraemic
shock was investigated. Human umbilical vein endothelial cells (HUVECs) were cultured by
trypsin digestion method. TF activity was measured in the lysates of HUVEC by using a single step clotting assay. Specific
mRNA expression was detected by Northern blotting. In order to evaluate a possible contribution of the nuclear factor (
NF)-kappa B pathway on the effects observed, electrophoretic mobility shift assays (EMSA) were performed using nuclear extracts from HUVECs and
NF-kappa B-binding
oligonucleotides. The results showed that treatment of HUVEC with LPS resulted in a significant increase in TF activity.
Anisodamine dose-dependently inhibited LPS-induced upregulation of TF. These effects was also confirmed on the level of specific TF
mRNA expression by Northern blotting. Furthermore, EMSA showed that
anisodamine completely abolished LPS-induced
NF-kappa B DNA binding activity in nuclear extracts from HUVECs treated with LPS together with
anisodamine. The results suggest that
anisodamine counteracts endothelial cell activation by inhibiting LPS-induced TF expression in these cells. Its interference with the
NF-kappa B pathway might--at least in part--contribute to this effect. The ability of
anisodamine to counteract LPS effect on endothelial cells might be one underlying mechanism explaining its antithrombosis and efficacy in the treatment of bacteraemic
shock.