Pregnancy induces expression of cPLA2 in ovine uterine artery but not systemic artery endothelium.

Abstract

To determine whether pregnancy increases cytosolic phospholipase A2 (cPLA2) expression in uterine artery (UA) endothelial cells and vascular smooth-muscle (VSM) cells, and whether ovarian steroids mediate this effect. Uterine arteries and omental arteries (systemic control) were isolated from pregnant (120-130 days' gestation) ewes and from nonpregnant ewes synchronized to the corresponding phases of the ovarian cycle (follicular or luteal) or ovariectomized. In addition, ovariectomized ewes were treated with vehicle, estradiol-17 beta (E2 beta), progesterone (P4), or combined E2 beta/P4 for 10 days, and UAs were collected. Arteries from all studies were separated into mechanically isolated endothelial and VSM fractions. Proteins were then solubilized and separated on 7.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western immunoblotting using an affinity-purified mouse monoclonal antibody for cPLA2. Both UA endothelium and VSM fractions were seen to express cPLA2, detected as a single band with a molecular mass similar to that observed in myometrium (about 100 kD). Distribution of cPLA2, when expressed per microgram of protein, was observed as 40% in UA endothelium compared with 60% in VSM fractions. Uterine artery endothelial cPLA2 expression was specifically increased 1.9-fold in pregnancy (P < .05), whereas there was no significant change from VSM. Furthermore, in ovariectomized sheep versus intact luteal or follicular phase animals, there was no significant change in cPLA2 expression in endothelium or VSM. Administration of E2 beta, P4, or their combination in ovariectomized sheep also failed to reproduce the pregnancy-induced increase in cPLA2 expression in UA endothelium. Omental artery endothelial and VSM cPLA2 expression was observed at similar magnitudes as UA expression, but levels were consistently unchanged by pregnancy, the ovarian cycle, or ovariectomy. cPLA2 was expressed throughout the endothelium and VSM of both uterine and omental arteries, but only in UA endothelium was pregnancy associated with elevated cPLA2 expression. The lack of change in cPLA2 expression with the ovarian cycle or in ovariectomized animals, even after prolonged treatment with E2 beta, P4, or E2 beta/P4 combined suggests this pregnancy-induced increase in cPLA2 may not be estrogen and/or progesterone dependent.