Mechanism of uterine vascular refractoriness to endothelin-1 in pregnant sheep

Endothelin-1 (ET-1) is a potent vasoconstrictor and produces marked pressor responses when given systemically. Studies in sheep have demonstrated that during pregnancy the uterine vasculature is refractory to exogenously administered ET-1. We hypothesize that this pregnancy-dependent refractoriness is due to an upregulation of local uterine metabolism of ET-1 and/or ET(B) receptors and/or downregulation of local uterine ET(A) receptors. To investigate these possibilities, 21 nonpregnant and 17 pregnant sheep were used. Dose-response curves to intravenous infusion of ET-1 and phenylephrine were generated for pregnant and nonpregnant sheep. ET-1 infused systemically demonstrated vasoconstriction in the systemic and renal vasculature of pregnant and nonpregnant animals and vasoconstriction in the uterine vasculature of nonpregnant animals. The pregnant animals showed no uterine vascular response to ET-1. In contrast, phenylephrine showed vasoconstriction in the systemic, renal, and uterine circulations in both pregnant and nonpregnant sheep. After experimentation, the animals were euthanized, and tissues were harvested for Western blot and activity analysis of neutral endopeptidase (NEP) or RT-PCR analysis of endothelin-converting enzyme (ECE) and ET(A) and ET(B) receptors. The content and activity of NEP in the uterine and renal vasculature of pregnant and nonpregnant animals were similar. RT-PCR demonstrated the presence of ECE in the uterine vasculature of pregnant and nonpregnant sheep. ET(A) receptor mRNA was significantly reduced in pregnant compared with nonpregnant sheep, whereas ET(B) receptor mRNA remained unchanged. We conclude that the uterine vascular refractoriness seen in the pregnant sheep is due to a downregulation of ET(A) receptors.