Release of Type II phospholipase A2 immunoreactivity and phospholipase A2 enzymatic activity from human placenta.
The aim of this study was to determine whether Type II phospholipase A2 (PLA2) is released from late pregnant human placental tissue. Placental explants were incubated in vitro and the release of immunoreactive (ir) Type II PLA2 and PLA2 enzymatic activity into the medium was determined. Both irType II PLA2 and PL2 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 PLA2 and PLA2 enzyme activity present in incubation medium were significantly correlated (P < 0.01). Consistent with the hypothesis that Type II PLA2 may be store in secretory granules within human placental tissue, incubation in the presence of a membrane depolarising concentration of KCI (60 mM) caused the release of irType II PLA2 2.0-fold (P < 0.001). PLA2 enzyme activity released into the incubation medium displays biochemical characteristics consistent with those previously reported for secretory PLA2 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 PLA2 is released from term placenta, when incubated in vitro. The release of this extracellularly-active PLA2 isozyme may contribute to gestational and labour-associated increases in glycerophospholipid metabolism and prostaglandin formation.
Farrugia
W
W
Department of Perinatal Medicine, Royal Women's Hospital, Carlton, Victoria, Australia.
Rice
G E
GE
Wong
M H
MH
Scott
K F
KF
Brennecke
S P
SP
D005260:Female; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D009924:Organ Culture Techniques; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D011263:Pregnancy Trimester, Third; D013997:Time Factors
D007527:Isoenzymes; D010741:Phospholipases A; D054467:Phospholipases A2