Prostacyclin synthetic activity in cultured aortic endothelial cells undergoing cyclic mechanical deformation

The effect of mechanical stretching on prostacyclin (PGI2) synthesis was studied by growing bovine aortic endothelial cells on flexible-bottomed culture plates that could be deformed by vacuum. A stress unit was used to exert an elongation of 24% at maximum downward deflection of the culture plate bottom. The experimental group was subjected to cycles of 10 seconds of elongation, 10 seconds of relaxation for 1, 3, or 5 days. The control group was subjected to similar incubations but without cyclic stretch. Twenty-four hours before collection, the medium was replaced with new medium that was devoid of serum. On days 1, 3, and 5, the 24-hour culture medium was collected (basal state). Arachidonic acid (20 mumol/L) was then added to each culture and incubated for 5 minutes at 37 degrees C. The medium was then collected to assess prostacyclin synthetic activity (stimulated state). Media were assayed for PGI2 and thromboxane A2 by radioimmunoassays for 6-keto-PGF1 alpha and thromboxane B2, the respective stable hydrolysis product. The results indicate that cyclic stretching, while not altering the basal production of PGI2, increases PGI2 synthetic capacity in a time-dependent manner. These data suggest that it may be inappropriate to extrapolate the mechanisms of in vivo PGI2 release from studies of endothelial cells in stationary culture.