Release of type II phospholipase A(2) immunoreactivity and phospholipase A(2) enzymatic activity from human placenta

The aim of this study was to determine whether Type II phospholipase A(2) (PLA(2)) is released from late pregnant human placental tissue. Placental explants were incubated in vitro and the release of immunoreactive (ir) Type II PLA(2) and PLA(2) enzymatic activity into the medium was determined. Both irType II PLA(2) and PLA(2) enzymatic activity accumulated in the incubation medium in a time-dependent manner (P<0.0001). This release was not associated with a loss of cell membrane integrity, as indicated by measurement of the intracellular enzyme, lactate dehydrogenase, in the incubation medium. The concentration of irType II PLA(2) and PLA(2) enzyme activity present in incubation medium were significantly correlated (P<0.01). Consistent with the hypothesis that Type II PLA, may be stored in secretory granules within human placental tissue, incubation in the presence of a caused the release of irType II PLA(2) 2.0-fold (P<0.001). PLA(2) enzyme activity released into the incubation medium displays biochemical characteristics consistent with those previously reported for secretory PLA(2) isozymes, that is, a requirement for millimolar concentrations of calcium for optimal enzyme activity, inhibited by reducing agents, such as dithiothreitol and insensitive to heat inactivation. The data obtained in this study establish that irType II PLA(2) is released from term placenta, when incubated in vitro. The release of this extracellularly-active PLA(2) isozyme may contribute to gestational and labour-associated increases in glycerophospholipid metabolism and prostaglandin formation.