Aim To investigate the effects and mechanisms of anisodamine on plasminogen activator inhibitor type 1 (PAI 1) expression in normal endothelial cells (EC) and lipopolicaccharide (LPS) treated EC. Methods Human umbilical vein endothelial cells (hUVEC) were cultured by trypsin digestion method. PAI 1 and tissue plasminogen activator (tPA) proteins in hUVEC conditioned medium were measured by sandwich enzyme linked immunosorbent assay (ELISA), and their mRNA expressions were determined by Northern blot analysis. Using electrophoretic mobility shift assay (EMSA), we assessed hUVEC nuclear factor kappa B (NF κB) nuclear translocation. Results LPS treatment of cultured hUVEC resulted in a significant increase in PAI 1 protein as well as mRNA expression by these cells. However, when hUVEC were incubated with LPS plus anisodamine, the upregulation of PAI 1 by LPS was abated. Moreover anisodamine was able to decrease the basal level of PAI 1 protein and mRNA as compared to control. Nuclear extracts prepared from LPS stimulated hUVEC demonstrated increased binding to the NF κB oligonucleotide as compared to unstimulated cells, and anisodamine could block those binding in the presence of LPS. Conclusion Anisodamine downregulated both basal and LPS induced PAI 1 protein and mRNA expression in EC, and the mechanism of the modulation might be via NF κB pathway.