Human equilibrative nucleoside transporters 1 and 2 may be differentially modulated by A(2B) adenosine receptors in placenta microvascular endothelial cells from pre-eclampsia

Abstract
Pre-eclampsia is associated with elevated maternal blood pressure and proteinuria, altered fetal growth, and increased plasma adenosine concentration in the mother and the fetus. Human equilibrative nucleoside transporters 1 (hENT1) and hENT2 are crucial to maintain physiological plasma levels of adenosine, thus modulating its several biological effects through adenosine receptor activation. However, it is unknown whether hENTs and adenosine receptors are expressed in human placental microvascular endothelium (hPMEC). To assay whether the increased fetal plasma adenosine concentration in pre-eclampsia results from altered hENT-mediated transport, and the potential involvement of adenosine receptors in this phenomenon, we investigated hENTs and A(2A) and A(2B) adenosine receptors expression and function in hPMEC. Cells were isolated and Cultured from normal pregnancies (n = 17) or Pre-eclampsia with adequate-for-gestational age fetuses (n = 7). hENT1, hENT2, A(2A) and A(2B) adenosine receptors were expressed and functional in hPMEC. Extracelhilar adenosine concentration was higher (4-fold) in pre-eclampsia versus normal pregnancies. hPMEC from pre-eclampsia exhibit increased total ;transport (hENT1 + hENT2), and maximal velocity (V-max) for hENT2- (2-fold), but reduced V-max for hENT1-mediated adenosine transport (75%), with no changes in apparent K-m. hENT2 expression was increased (4.5-fold), but hENT1 protein abundance was reduced (80%) in pre-eclampsia. Equally, A2A expression was reduced (50-80%) in pre-eclampsia. CGS-21680 (A(2A) agonist) did not alter hENTs expression or activity, but ZM-241385 (A(2A) antagonist) blocked pre-eclampsia effects and increased hENT1-mechated transport in normal pregnancies. Thus, A(2B) adenosine receptors may differentially modulate hENTs in hPMEC, which could be a mechanism attempting to re-establish physiological extracellular adenosine levels in pre-eclampsia. (C) 2008 Elsevier Ltd. All rights reserved.
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Keywords
human, placenta, adenosine, endothelium, pre-eclampsia, UMBILICAL VEIN ENDOTHELIUM, SLC29A1 PROMOTER ACTIVITY, FETAL-GROWTH RESTRICTION, ADENOSINE TRANSPORT, PLASMA ADENOSINE, NUCLEOSIDE, RECEPTORS, PREGNANCY, EXPRESSION, VASOCONSTRICTION
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