Estrogen stimulates von Willebrand factor production by cultured endothelial cells

Monolayers of cultured human umbilical vein endothelial cells were exposed to 17 beta-estradiol and compared to control cultures with respect to levels of von Willebrand factor (vWF) released into the media after 3-5 days of incubation. The amount of functional vWF activity was assessed by ristocetin-induced platelet aggregation and by a radioreceptor platelet assay. vWF antigen was quantitated by immunoassay. The DNA content of each monolayer was determined fluorometrically and used as a measure of cell number. By all assays, vWF levels in the media from the estradiol-treated endothelial cells were reproducibly and significantly higher when compared with control values. The amount of vWF produced by the cultured endothelial cells showed a dose-response effect to the estradiol added to the media. The estradiol-treated cells produced approximately 1.3 +/- 0.30 micrograms vWF/ml/micrograms DNA at 2 ng estradiol/ml, compared with control cultures that produced 0.75 +/- 0.16 microgram vWF/ml/micrograms DNA (p less than 0.001). The estradiol-treated monolayers consistently contained slightly greater amounts of DNA than control cultures: 2.0 +/- 0.10 micrograms versus 1.7 +/- 0.12 micrograms DNA (p less than 0.001). By multivariant analysis, however, the differences in cell number could only account for less than or equal to 10% of the elevation in the level of vWF that occurred in response to estradiol. By SDS-agarose electrophoresis and radioimmunoblotting, the vWF within the cytosol of the endothelial cells was found to possess a multimeric pattern similar to that found for either purified plasma vWF or vWF released into media overlying endothelial cell cultures. Our studies indicate that estrogen directly stimulates endothelial cells to increase their rate of production of vWF and, in addition, causes a slight increase in endothelial cell replication. These data may bear on the observation that administration of estrogen to some women with von Willebrand's disease causes an increase in their functional levels of vWF.