dinoprost and arachidonyltrifluoromethane

We investigated role(s) of luteal group IVA phospholipase A(2) (GIVA PLA(2)) in prostaglandin (PG) F(2alpha)-induced regression in pseudopregnant rats. Prostaglandin F(2alpha) (PGF(2alpha)) treatment of day 6 pseudopregnant rats stimulated luteal PLA(2) activity, which was sensitive to inhibitors and associated with increased GIVA PLA(2) immunoreactivity. Intra-bursal treatment with the enzyme inhibitor (AACOCF3) prior to PGF(2alpha) failed to prevent the initial decline in progesterone but induced subsequently a persistent rise that was significantly higher than that of vehicle-treated group. TUNEL-positive signals in luteal cells of control group were reduced by AACOCF3 treatment. TUNEL-positive reaction induced in luteal cells in vitro by combined cytokines and agonistic anti-Fas were both reduced by AACOCF3 and another inhibitor pyrrophenone. Overall data show that luteal GIVA PLA(2) activity and expression increased following PGF(2alpha) administration and that acute chemical inhibition of this activity could reverse, at least partly, PGF(2alpha)-induced functional regression and prevent apoptosis induced by PGF(2alpha)in vivo and by cytokines in vitro.

Topics: Animals; Apoptosis; Arachidonic Acids; Dinoprost; Enzyme Inhibitors; Female; Immunohistochemistry; In Situ Nick-End Labeling; Luteolysis; Male; Phospholipases A2; Progesterone; Pseudopregnancy; Pyrrolidines; Rats; Rats, Wistar

Changes in endothelium functions during ischemia are thought to be of importance in numerous pathological conditions, with, for instance, an increase in the release of inflammatory mediators like prostaglandins. Here, we showed that hypoxia increases phospholipase A(2) (PLA(2)) activity in human umbilical vein endothelial cells. Both basal PLA(2) activity and PG synthesis are sensitive to BEL and AACOCF3, respectively, inhibitors of calcium-independent PLA(2) (iPLA(2)) and cytosolic PLA(2) (cPLA(2)), while OPC, an inhibitor of soluble PLA(2) (sPLA(2)) only inhibited the hypoxia-induced AA release and PGF(2alpha) synthesis. Hypoxia does not alter expression of iPLA(2), sPLA(2) and cPLA(2) and cycloheximide did not inhibit PLA(2) activation, indicating that hypoxia-induced increase in PLA(2) activity is due to activation rather than induction. However, mRNA levels for sPLA(2) displayed a 2-fold increase after 2 hr incubation under hypoxia. BAPTA, an intracellular calcium chelator, partially inhibited the AA release in normoxia and in hypoxia. Direct assays of specific PLA(2) activity showed an increase in sPLA(2) activity but not in cPLA(2) activity after 2hr hypoxia. Taken together, these results indicate that the hypoxia-induced increase in PLA(2) activity is mostly due to the activation of sPLA(2).

Topics: Arachidonic Acid; Arachidonic Acids; Cell Hypoxia; Cells, Cultured; Dinoprost; Endothelium, Vascular; Enzyme Inhibitors; Humans; Isoenzymes; Naphthalenes; Phospholipases A; Phosphorylcholine; Pyrones

Although transforming growth factor alpha (TGF-alpha) is known to be an important survival factor for granulosa cells, the cellular and molecular mechanisms involved are uncertain. The purpose of the present study was to investigate the possible involvement of prostaglandins in the anti-apoptotic action of TGF-alpha. Hen granulosa cells from healthy prehierarchical follicles (2-6 mm) cultured in serum-free medium underwent spontaneous apoptosis as demonstrated by DNA fragmentation and nuclear chromatin condensation. TGF-alpha (20 ng ml(-1)) stimulated maximum synthesis of prostaglandins (PGE and PGF) in granulosa cells and completely inhibited serum deprivation-induced apoptosis. The addition of an inhibitor of cyclooxygenase (COX; N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide (NS398) or ibuprofen) or phospholipase A(2) (PLA(2); aristolochic acid, 2-p-amylcinnamoyl amino-4-chlorobenzoic acid (ONO-RS-82) or arachidonyl triflouro methyl ketone (TFMK)), to the culture medium markedly suppressed the TGF-alpha-induced prostaglandin synthesis and significantly increased granulosa cell apoptosis. The apoptotic effect of NS398 and aristolochic acid was completely inhibited by exogenous prostaglandins (PGF(2 alpha), PGE(1), PGE(2)) and arachidonic acid, respectively. However, exogenous prostaglandins failed to inhibit the PLA(2) inhibitor-induced apoptotic DNA fragmentation, implying that in addition to prostaglandins, arachidonic acid or leukotrienes may be important in transducing the anti-apoptotic action of TGF-alpha. In the absence of exogenous TGF-alpha, prostaglandins had no significant influence on granulosa cell apoptosis induced by serum withdrawal. These findings indicate that prostaglandin synthesis is a necessary, but not sufficient, event in the suppression of granulosa cell apoptosis by TGF-alpha. Whether arachidonic acid or leukotrienes are important in the anti-apoptotic action of TGF-alpha in hen granulosa cells remains to be determined.

Topics: Alprostadil; Animals; Apoptosis; Arachidonic Acid; Arachidonic Acids; Aristolochic Acids; Cells, Cultured; Chickens; Chromatin; Culture Media, Serum-Free; Cyclooxygenase Inhibitors; Dinoprost; Dinoprostone; DNA Fragmentation; Enzyme Inhibitors; Female; Granulosa Cells; Ibuprofen; Nitrobenzenes; Phenanthrenes; Phospholipases A; Prostaglandins; Prostaglandins E; Prostaglandins F; Sulfonamides; Transforming Growth Factor alpha