Lipocortin-I inhibits the synthesis and release of prolactin from human decidual cells: evidence for autocrine/paracrine regulation by lipocortin-I.
The lipocortins are a family of calcium-dependent phospholipid-binding proteins that are induced by glucocorticoids and inhibit phospholipase-A2 activity. To determine whether the lipocortins affect the release of PRL from human decidua, decidual cells from term pregnancies were exposed to recombinant lipocortin-I for 96 h, with medium changes at 24-h intervals. Lipocortin-I (0.01-100 nM) caused a time- and dose-dependent inhibition of PRL release, with a half-maximal effective dose of 50 nM. PRL release was inhibited by 27%, 62%, 93%, and 98% at 24, 48, 72, and 96 h, respectively. The cells exposed to lipocortin-I did not release the enzymes alkaline phosphatase and lactic dehydrogenase, indicating that the inhibitory effect on PRL release was not due to cell death. In addition to inhibiting basal PRL release, lipocortin also completely inhibited the stimulation of PRL release by decidual PRL-releasing factor, a 23.5-kDa protein recently purified from human placenta that stimulates the synthesis and release of decidual, but not pituitary, PRL. Hydrocortisone and dexamethasone (0.1-10 microM) had no effect on PRL release, and arachidonic acid (2-100 microM) inhibited rather than stimulated PRL release. Western blot analysis demonstrated the presence of lipocortin-I in decidual cells and conditioned media. On Northern blot, decidual mRNA hybridized to an oligonucleotide for lipocortin-I. These results strongly suggest that lipocortin-I has an autocrine/paracrine role in regulation of the synthesis and release of PRL from human decidual cells.
Pihoker
C
C
Department of Pediatrics, Duke University Medical Center, Durham, North Carolina 27710.
Feeney
R J
RJ
Su
J L
JL
Handwerger
S
S
D017302:Annexins; D015152:Blotting, Northern; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology; D003470:Culture Media; D003656:Decidua / Q000166:cytology / Q000378:metabolism*; D004586:Electrophoresis; D004702:Endocrine Glands / Q000378:metabolism*; D005260:Female; D006801:Humans; D009994:Osmolar Concentration; D010740:Phospholipases / Q000037:antagonists & inhibitors; D011388:Prolactin / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins
D017302:Annexins; D002135:Calcium-Binding Proteins; D003470:Culture Media; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011388:Prolactin; D010740:Phospholipases