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[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.
Conclusion:
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.