[A study on the mechanism of anisodamine against thrombosis].
OBJECTIVE
To investigate the effects and the mechanism of Chinese herb anisodamine on PAI-1 expression in LPS stimulated endothelial cells (ECs).


METHODS
Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion method. PAI-1 protein in HUVEC conditioned medium was measured by sandwich enzyme-linked immunosorbent assay (ELISA), and PAI-1 mRNA expression in HUVEC was determined by Northern blot analysis. HUVEC nuclear factor-kappa B (NF-kappaB) nuclear translocation was assessed by immunocytochemistry technique.


RESULTS
LPS treatment of cultured HUVECs resulted in a significant increase of PAI-1 protein as well as mRNA expression by these cells. However, when HUVECs were incubated with LPS plus anisodamine, the upregulation of PAI-1 by LPS was significantly abated. The expression of PAI-1 protein and mRNA was decreased by 30% and 24% respectively compared with LPS treatment alone. Moreover, anisodamine was able to decrease the basal level of PAI-1 protein and mRNA by 74% and 70% respectively. Furthermore, anisodamine could block LPS induced NF-kappaB of HUVECs nuclear translocation.


CONCLUSIONS
Anisodamine inhibits both basal and LPS-induced PAI-1 protein as well as mRNA expression in ECs. The inhibition of anisodamine on LPS-induced PAI-1 in ECs may take place via the NF-kappaB pathway.
Ruan
Q
Q
Department of Pathology, Tongji Medical University, Wuhan 430030, China.
Song
J
J
Deng
Z
Z
D002478:Cells, Cultured; D004365:Drugs, Chinese Herbal / Q000494:pharmacology; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012991:Solanaceous Alkaloids / Q000494:pharmacology*; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation / Q000187:drug effects; D014665:Vasodilator Agents / Q000494:pharmacology
D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D012991:Solanaceous Alkaloids; D014665:Vasodilator Agents; C003922:anisodamine