Anisodamine inhibits endotoxin-induced tissue factor expression in human endothelial cells.
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.
Ruan
Qiurong
Q
Department of Pathology, School of Basic Medical Sciences, Tongji Medical College.
Song
Jianxin
J
Deng
Zhongduan
Z
D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012991:Solanaceous Alkaloids / Q000494:pharmacology*; D013925:Thromboplastin / Q000096:biosynthesis / Q000235:genetics*; D014471:Umbilical Veins / Q000166:cytology
D004365:Drugs, Chinese Herbal; D008070:Lipopolysaccharides; D016328:NF-kappa B; D012333:RNA, Messenger; D012991:Solanaceous Alkaloids; C003922:anisodamine; D013925:Thromboplastin