Summary
To learn whether glucocorticoids inhibit prostaglandin (PG) production in vascular
endothelial cells, we investigated the effects of glucocorticoids on PG synthesis
by cultured human umbilical vein endothelial cells (EC). Pretreatment of EC with dexamethasone
(DX, 10-9 to 5 x 10-5 M) caused a dose-dependent inhibition of PGI2 production when PG synthesis from endogenous arachidonate was stimulated by human
thrombin (0.25-2 U/ml) or ionophore A 23187 (1-5 μM). The inhibition was detectable
at 10-7 M DX and maximal at 10-5 M (4.0 ± 0.7 vs. control: 7.7 ± 1.9 ng/ml, mean ± S.D., P <0.01). The production
of PGE2 and the release of radiolabelled arachidonate (AA) from prelabelled cells were similarly
inhibited. Prolonged incubation of EC with glucocorticoids was required to inhibit
PG production or arachidonate release: ranging from 8% inhibition at 5 h to 44% at
38 h. In contrast, prostaglandin formation from exogenous AA was not altered by DX
treatment. When thrombin or ionophore-stimulated EC were restimulated with exogenous
AA (25 μM), DX-treated cells released more PGI2 than control cells (5.7 ± 0.5 vs.
4.1 ± 0.6 ng/ml, P <0.01). Both the decrease in PGI2 production after thrombin/ionophore
and the increase after re-stimulation with AA were blunted in the presence of the
protein synthesis inhibitor cycloheximide (0.1-0.2 μg/ml). Thus, incubation of EC
with glucocorticoids inhibits PG production at the step of phospholipase activation.
The time requirement for these steroid effects and their blunting by cycloheximide
are consistent with the induction of regulatory proteins, possibly lipocortins, in
endothelial cells.
Keywords
Glucocorticoids - Endothelial cells - Prostaglandins - Prostacyclin - Phospholipases
