O-076 Video mapping to assess efficacy of an antiestrogen (raloxifene) on spontaneous endometriosis in the rhesus monkey

and prostgrandins (PGs) (metabolites of AA) in terms of PGV proliferation.Design: Effects of phospholipasa A2 (PLA2) inhibitor, PLC inhibitor, phosphatidic acid (PA) phosphohydrolase inhibitor, DAG lipase inhibitor, cycloxygenase inhibitor with or without AA and PGs on the PGV proliferation.Materials and Methods: Porcine GCs from medium-size follicles were infected by SV40, then cloned and cultured for more then 9 years.PGV were cultured in flasks with TCM199 containing 10% fetal bovine serum.After trypsin digestion, PGV (l×104/well) were inoculated to 96 wells.After 2 days of culture, various doses (10-12~10-~M) of PLA2 inhibitor (bromophenacyl bromide, BPB), PA phosphohydrolase inhibit (propranolol, PPL), PLC inhibitor (U73122, U), DAG lipase inhibitor (RHC80267, RHC), cycloxygenase inhibitor (indomethacin) with or without 10-12~10 -6 of AA and PGs (PGE1, E2, F2a, I2 and D2) were added to PGV with TCM199 containing 10% horse serum.After 2 days of culture with reagents, cell numbers were counted using absorptiometer with XTT.Results: BPB, PPL, U, RHC and indomethacin decreased PGV cell numbers dose-dependently.AA reversed the inhibition of PGV cell replication by BPB, U, PPL, and RHC.PGE1, E2, and F2a effectively increased PGV cell numbers to control levels when added with indomethacin.PG12 and D2 had no effects.Conclusions: These results confirm the central role of DAG in the proliferation of PGV and suggest that DAG is synthesied not only through the phospholipase C pathway from phosphatidyl inositol but also through the PLD pathway from other phospholipids.AA is synthesied from phospholipids through PLA2 and also from DAG.Our data also suggest that, besides PKC, AA (DAG metabolite) and PGs (AA metabolites) play an important role in the PGV proliferation.The mechanisms of these reagents must be studied further.