Influence of hydroxylation and conjugation in cross-inhibition of bile acid transport across the human trophoblast basal membrane

Abstract

Taurocholate (TC) transport across the basal plasma membrane of the human trophoblast is a carrier-mediated process, whose specificity is probably not restricted to TC. The aim of this work was to gain further insight into the role of hydroxylation and conjugation in the behavior of the carrier system vs. bile acid (BA) species. Radiolabeled TC transport by basal plasma membrane (BPM) vesicles obtained from human term placenta was measured by a rapid filtration technique. Glycocholate (GC), taurochenodeoxycholate (TCDC) and taurodeoxycholate (TDC) inhibited TC binding to BPM. These bile acids complete with TC for the binding sites. Symmetry properties for GC- and TCDC-induced inhibition of TC transport was found in experiments where GC or TCDC were at the cis-side of the membrane (uptake and efflux experiments). GC and TCDC-induced inhibition seems to be of mixed type. By contrast, TDC was observed to affect TC transport differently, depending on whether the experiments addressed uptake or efflux. At the intracellular side of the membrane (uptake), TDC induced a marked increase in both V max and K t . However, at the fetal side (efflux) a significant reduction in both V max and K t was found. In spite of these peculiarities, the values for K i were very close for GC, TCDC and TDC at the intracellular side but not at the fetal side, where the decreasing order for K i was GC > TCDC > TDC . TC uptake by BPM vesicles was not modified in the presence of a wide range of estrone sulfate concentrations (0.002–1.0 mM). In summary, these results indicate that a particular bile acid molecular structure is necessary for steroid-related compounds to interact with the bile acid carrier located in BPM. They also suggest that changes in the number and position of hydroxy groups, as well as in the amino-acid moiety in amidated bile acids modify the behavior of the carrier, which may play an important role in the net vectorial transfer of bile acids across the placenta.