The origin of circulating 13,14-dihydro-15-keto-prostaglandin F2α during delivery

All uterine tissues as well as the fetal membranes and the placenta can form prostaglandins from endogenous precursors but it is not clear which of the tissues is the main site for the increase in PGF_2α production during human parturition. To examine this question, we measured plasma prostaglandin levels before and at intervals after expulsion of the fetus, placenta, and membranes. The concentration of PGFM at the beginning of the second stage of labor was significantly higher than before the onset of labor. Five minutes after the birth of the infant, the concentration had doubled. Thirty minutes after the expulsion of placenta and membranes, plasma PGFM had fallen to the levels at full dilatation; two hours postpartum it was still significantly raised over levels before labor. Since the halflife of PGFM in the circulation is about 7 minutes, these findings indicate that the uterine tissues are important sources of PGFM during labor. In contrast, endogenous oxytocin levels, which were significantly raised over control levels at the second stage of labor, did not change during the third stage, and decline postpartum to control levels. Oxytocin infusion did not influence PGFM levels at 5 and at 30 minutes postpartum, but raised them at 2 hours.     

Jugular levels of 13,14-dihydro-15-keto-prostaglandin F and progesterone around luteolysis and early pregnancy in the ewe

Six non-pregnant ewes at day 12 of the estrous cycle each had a day-12 embryo transferred into the uterine horn ipsilateral to the corpus luteum, and 4 non-pregnant ewes at day 13 each had a day-13 embryo similarly transferred. Four control ewes, 2 at day 12 and 2 at day 13 received sheep serum into the uterine horn ipsilateral to the corpus luteum. Jugular blood samples were taken at 2-hourly intervals for 3 days post-surgery, then twice-daily for a further 4 days, and the plasma radioimmunoassayed for progesterone and 13,14-dihydro-15-keto-prostaglandin F. All control ewes exhibited estrus within the expected time range and pulsatile peaks of 13,14-dihydro-15-keto-prostaglandin F occurred coincident with declining progesterone levels. With one exception, the recipient ewes had prolonged cycles and those ewes found pregnant at necropsy, 30 days after transfer, showed no progesterone decline and no pulsatile peaks of prostaglandin during days 12 to 16 after estrus. These observations suggest that the presence of the embryo at a critical stage after mating suppresses the release of uterine prostaglandin F_2α.     

Temporal relationships between peripheral plasma concentrations of oxytocin, progesterone and 13, 14-dihydro-15-keto-prostaglandin F2α during the oestrous cycle and early pregnancy in the ewe

Peripheral plasma concentrations of oxytocin, 13,14-dihydro-15-keto-prostaglandin F_2α(PGFM), progesterone and LH were determined at 3 hourly intervals during the oesterous cycle (n = 3) and in early pregnancy (n = 4) in sheep. The progesterone and LH concentrations showed that the cycling ewes were samples during the periods of luteal regression (decreasing progesterone concentrations), the preovulatory gonadotrophin surge and the beginning of the next luteal phase (increasing progesterone concentrations). The pregnant ewes had basal LH concentrations and luteal phase concentrations of progesterone (>lng/ml afte day 5 following mating) throughout the whole of the sampling period. Oxytocin concentrations in the non-pregnant ewes decreased around the time of luteal regression to reach low concentrations (mean concentrations of approximately 18pg/ml) during the preovulatory period and then increased after the preovulatory surge. PGFM concentrations exhibited a pulsatile pattern with increasing concentrations as progesterone levels fell. In the pregnant ewes oxytocin concentrations gradually fell until approximately 16 days post-mating (approximately 7–8pg/ml). The magnitude of the pulses in PGFM concentrations were also lower than in the cycling ewes. These results demonstrate that the increased concentrations of PGFM which are found during the period of luteal regression are not caused by increased peripheral concentrations of oxytocin.     

Plasma levels of 13,14-dihydro-15-keto prostaglandin F2α, estrogens, and progesterone during stretch-induced labor at term

The uterus of six healthy multiparous women at term was mechanically stretched by a rubber catheter and balloon. Apparent labor was inaugurated in all cases within 5 hours and increased progressively with time. Advanced cervical softening and dilatation were also evident after the stretch treatment. Significant increases in the levels of 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM) were observed with the progress of treatment (P < 0.01). Plasma estrogens and progesterone levels did not change significantly during the treatment (P > 0.05). Stretching and/or resulting uterine contractions appear to induce the secretion of prostaglandin F_2α (PGF) from the organ, which in turn seems to be involved in both cervical softening, and the onset and progress of labor, under stable conditions of plasma estrogens and progesterone.     

Effects of dibuturyl cAMP, LHRH, and aromatase inhibitor on simultaneous outputs of prostaglandin F2α, and 13,14-dihydro-15-keto-prostaglandin F2α by term placental explants

Explants from term human placentas were maintained in culture with daily changes of medium. Daily output of PGF_2α and PGFM¹ decreased during the course of the incubation. Addition of 4 μg/ml DHEAS or 67 μg/ml LDL cholesterol had no effect on output of PGF_2α or PGFM. Addition of 1.6, 3.2, or 6.4 μg/ml of LHRH to the culture plates had no effect on output of PGFM or PGF_2α, but LHRH increased hCG output. Dibutyryl cAMP (1mM, 2mM, and 4mM) increased output of PGF_2α, PGFM, and hCG. Aromatase inhibitor decreased hCG output, but it was without effect on output of PGF_2α, or PGFM. Significant correlations were demonstrated between progesterone, PGFM, PGF_2α, and hCG, suggesting that PGF_2α originates in the syncytiotrophoblast cell. The ability of LHRH to stimulate output of hCG but not PGF_2α while dbcAMP stimulated both suggests that either PGF_2α and hCG arise in different cells or that LHRH does not act through cAMP.     

Influence of low plasma progesterone concentrations on the release of prostaglandin F2α during luteolysis and oestrus in heifers

A study was conducted to determine the effect of suprabasal plasma concentrations of progesterone on the release of prostaglandin F_2α (PGF_2α) at luteolysis and oestrus. Heifers received silicone implants containing 2.5 (n = 4), 5 (n = 4), 6 (n = 3), 7.5 (n = 3), 10 (n = 4), or 15 (n = 3) g of progesterone, or an empty implant (controls, n = 4) between Days 8 and 25 post ovulation. Blood was collected frequently between Days 14 and 28 and assayed for progesterone and 15-ketodihydroprostaglandin F_2α. Basal progesterone concentrations in control heifers did not differ from those in heifers with 2.5- or 5-g implants and remained around 0.4−0.5 nmol l⁻¹ until ovulation in all three groups. In the heifers treated with 6–15 g of progesterone, basal concentrations were maintained at higher (P < 0.05) levels compared with those in the controls, ranging from 0.8 to 1.6 nmol 1⁻¹. The effect of these elevated progesterone levels was to delay ovulation by prolonging the growth of the ovulatory follicle, which continued growing until the implant was removed. In all experimental groups, the first significant increase of the PGF_2α metabolite occurred between Days 15.3 and 16.3 (P > 0.05) and was associated with the onset of a decrease in progesterone concentrations, which had reached levels below 3 nmol 1⁻¹ by Days 17.4−19.1. PGF_2α metabolite peaks associated with luteolysis were frequent until Day 20. In the period from Day 20 until implant removal, sporadic peaks were observed, ranging in number from 1.0 ± 1.2 (mean ± SEM) in the control group to 3.0 ± 1.4 peaks in the heifers treated with 7.5 g of progesterone (P > 0.05). The number of PGF_2α metabolite peaks during that period was higher (P < 0.05) in heifers treated with 10 and 15 g than in controls. A positive correlation was found between the basal concentration of progesterone and the number of PGF_2α peaks after luteolysis (r = 0.54; P < 0.01). Plasma progesterone concentrations above approximately 1.4 nmol l⁻¹ were able to maintain the release of PGF_2α until the progesterone implants were removed and plasma levels decreased to basal values. These heifers had a preovulatory PGF_2α release pattern resembling that found in repeat breeder heifers.     

Temporal relationship between plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F and neurophysin I/II around luteolysis in sheep

Plasma concentrations of neurophysin I/II (N-I/II), 13,14-dihydro-15-keto-prostaglandin F (PGFM) and progesterone were measured by radioimmunoassay in plasma samples collected from four sheep at hourly intervals between 0700 and 1900 h from Days 12–17 of the estrous cycle. Plasma samples were also collected from a fifth sheep at 2-hourly intervals during Days 12–16 of the cycle. In all sheep, intermittent surges in the plasma concentrations of PGFM and N-I/II occurred during the period of luteal regression. On at least one occasion in each sheep a surge in the plasma concentration of N-I/II was observed coincident with a rise in PGFM concentrations. In general, the highest levels of N-I/II were observed early in luteolysis (Days 13–14 of the cycle) while the corresponding levels of PGFM in plasma were maximal around Day 15 when luteolysis was well advanced.It is suggested from this temporal data that oxytocin, which is considered to be released in association with N-I/II, may play an important role in ovine luteolysis by stimulating the secretion of prostaglandin F from the uterus during Days 13–15 of the estrous cycle.     

Changes in urinary 6-keto-prostaglandin F1α excretion during pregnancy and labor

Urinary excretion of 6-keto-PGF_1α was measured by high pressure liquid chromatography and radioimmunoassay at various stages of pregnancy and labor. In the first trimester of pregnancy, urinary 6-keto-PGF_1α concentrations were nor different from those measured before pregnancy, but they showed a significant increase in the second trimester of pregnancy (p <0.001). The levels rose further in the third trimester, although this increase was not statistically significant when compared to levels obtained in the second trimester. There was no evidence for a significance change in 6-keto-PGF_1α excretion with the onset of labor. During well-established, progressive labor mean values of 6-keto-PGF_1α excretion were about twice as high as before the onset of labor, but the range of values during labor was so wide that there was no statistical difference with values obtained in the second half of pregnancy.It is concluded that the increase in the urinary excretion of 6-keto-PGF_1α occurs later in pregnancy than the increase in TXB₂ excretion and that labor at term is not associated with marked changes in 6-keto-PGF_1α excretion.     

Plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F during pregnancy in sheep

A specific and sensitive radioimmunoassay is described for 13,14-dihydro-15-keto-prostaglandin F in ovine plasma. Using this assay it has been shown that, in sheep, jugular venous 13,14-dihydro-15-keto-prostaglandin F concentrations increase at parturition and correlate well with concentrations of prostaglandin F in the utero-ovarian vein. It is suggested that uterine prostaglandin F production under these conditions may be assessed by measuring peripheral venous 13,14-dihydro-15-keto-prostaglandin F, thereby avoiding the need for chronic utero-ovarian venous catheters.     

Effect of oxytocin on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha during the oestrous cycle and early pregnancy in the goat.

The effect of subcutaneous oxytocin on plasma concentrations of 13,14-dihydro-15-keto-prostaglandin (PG) F2 alpha (PGFM) was examined in the goat at various periods during the oestrous cycle and early pregnancy. 100 i.u. oxytocin was administered daily for 4 day, the dose being divided and given at 0900 and 2100 h; PGFM concentrations were assessed after the first treatment of each day. On days 3-6 (oestrus, day 0) PGFM concentrations increased significantly (P less than 0.001) within 15 minutes and both non-pregnant and mated goats exhibited oestrus behaviour by day 7. Significant (P less than 0.01) increases in PGFM were also produced on days 7-10, in both non-pregnant and pregnant goats, but the responses diminished from day 7 to day 10; only one goat (non-pregnant) came into oestrus. There was a marked difference in response between groups, however, during days 12-15. In non-pregnant goats significant (P less than 0.05) increases in PGFM were detected on days 13-15, but in pregnant animals oxytocin was without effect. Similarly, oxytocin did not increase PGFM concentrations on days 17-20 of pregnancy. However, uterine responsiveness reappeared in pregnant goats with significant (P less than 0.01) increases in PGFM on days 24 and 25.     

Intrauterine instillation of prostaglandin F2α in early pregnancy

Intrauterine PGF_2α (5mg) was administered for termination of early pregnancy in 14 healthy volunteers. With 11 complete abortions, the efficiency rate of this technique is below conventional methods. In addition, the incidence of infection was high occurring in 12 out of 14 subjects. Because of persistent bleeding, six patients underwent a dilatation and curettage. Other significant side effects included transient hypertension, pain, nausea and restlessness. In the patients with a complete abortion, the mean plasma progesterone concentration fell 37% after 8 hours post PGF_2α instillation and 90% 14 days later. The mean plasma estradiol-17β fell 26% over the initial eight hour period and 75% over the next 14 days.     

Effects of prostaglandins, dibutyryl camp, LHRH, estrogens, progesterone, and potassium on output of prostaglandin F2α, 13, 14-dihydro-15-keto-prostaglandin F2α, HCG, estradiol, and progesterone by placental minces

In order to compare the endocrine response of placental minces to luteinizing hormone releasing hormone (LHRH) and dibutyryl cAMP (dbcAMP) and to screen for effects of potential stimulatory and inhibitory substances, the simultaneous outputs of PGF_2α, 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM), progesterone, 17β-estradiol, and hCG were evaluated during a 4 hour incubation in 5 placentas. The output of hCG was highest for 12-week placentas, intermediate for a 16 week placenta, and lowest for term placentas. The output of 17β-estradiol by 12 and 16 week placentas in the presence of 30 μM dehydroepiandrosterone sulfate (DHEAS) was greater than that by term placentas. Progesterone output was apparently independent of gestational age although some variation between 12-week placentas was demonstrated. Output of PGF_2α was lower in 12 and 16-week placentas than in term placentas and that of PGFM was lower in 12-week placentas than in term placentas. LHRH (100 nM) produced stimulation of PGF_2α output (P<.005) and a trend toward inhibition of progesterone output (which failed to achieve statistical significance) but no stimulation of hCG under these conditions. Stimulation of the outputs of hCG (P<.005) and PGF_2α (P<.001) and inhibition of that of progesterone (P<.005) was produced by 20 mM dbcAMP. DHEAS inhibited output of progesterone (P<.01) and PGF_2α (P<.01). There were no effects of potassium, estrogens, progesterone, or prostaglandins on output of any measured substance.     

Disappearance of prostaglandins F2α and 15-methyl F2α from amniotic fluid as measured by radioimmunoassay

A sensitive and relatively specific radioimmunoassay for 15 (S) 15 methyl prostaglandin F_2α was used to determine the levels of the drug in amniotic fluid after it had been injected intra-amniotically for termination of second trimester pregnancy. The disappearance of the free acid (tham salt) and methyl ester of the prostaglandin analogue were similar. The results of this preliminary study suggest that the drug rapidly equilibrates in the fluid and this is followed by a slow removal from the amniotic sac. A comparison with a similar study with PGF_2α, revealed that the analogue had a longer half-life in the amniotic fluid.     

Conditions of the cervix for the increase of plasma levels of 13, 14-dihydro-15-keto-prostaglandin F2α after amniotomy at term

Amniotomy was performed in 12 multiparas at term but not in labor. In 6 of these patients (group I), the fetal head and cervix condition were favorable for amniotomy, and in the other 6 (group II), they were not favorable. In all group I patients, a sudden and progressive descent of the fetal head, and onset and progress of labor were noted within 5 hours. Plasma 13,14-dihydro-15-keto-prostaglandin F_2α (PGFM) levels increased significantly (P < 0.05)_in 4 of these cases with time. In group II patients, descent of the head was less than that in group I patients (P < 0.05), and neither strong labor nor rise of PGFM levels was noted within 5 hours. These data support our view that amniotomy at an appropriate time results in the onset and progress of labor, and the rise of plasma PGFM in virtue of the sudden and exponential increase of the head to cervix force, but amniotomy at an inappropriate time does not, because this force is unchanged.     

Elevated concentrations of 13,14-dihydro-15-keto-prostaglandin F-2 alpha in maternal plasma during prepartum luteolysis and parturition in dogs (Canis familiaris)

Concentrations of progesterone and of 13,14-dihydro-15-keto-prostaglandin F-2 alpha (PGFM) were measured in plasma collected from 6 bitches every 3 h starting 2.8-4.6 days before parturition (birth of first pup) and continuing until 0.4-0.8 days post partum, and in additional samples collected less frequently. Progesterone concentrations at 48, 24, 12 and 3 h pre partum averaged 2.8 +/- 0.3, 2.2 +/- 0.4, 1.0 +/- 0.3 and 0.7 +/- 0.2 ng/ml. At those times PGFM values averaged 380 +/- 80, 800 +/- 220, 1450 +/- 450 and 1930 +/- 580 pg/ml, respectively. Mean concentrations of PGFM increased about 2.5-fold between 48 and 15 h pre partum in association with the onset of luteolysis, and then increased another 2.5 times before parturition as progesterone fell to nadir values. Peak levels of PGFM ranged from 1060 to 7150 pg/ml (2100 +/- 600 pg/ml) and occurred within 1-9 h after the birth of the first pup and before the birth of the last pup. These results suggest that prepartum luteolysis in dogs is initiated by increases in maternal concentrations of PGF, and that progesterone withdrawal causes a further increase in PGF which completes luteolysis and provides a major portion of the uterotonic activity causing expulsion of pups.     

Cardiovascular actions of prostaglandins F2α; 15-me-F2α; F1α; F2β and F1β in the cat

Prostaglandin F_2α (5μg/kg, i.v.) causes an increase in pulmonary arterial pressure, decrease in systemic arterial pressure, and reflex bradycardia in the anesthetized cat. The same dose of the 15-methyl analogue of PGF_2α produces the same triad of effects but of greater magnitude and duration. Although prostaglandins F_1α, F_2β and F_1β also cause the same cardiovascular effects as F_2α, there is a decrease in potency for all parameters measured, with PGF_2α>PGF_1α>PGF_2β>PGF_1β. When compared to the actions of PGF_2α in producing an increase in pulmonary arterial pressure, PGs F_1α, F_2β and F_1β were less potent by approximately 10, 100, and 1000 fold respectively.     

Corpus luteum susceptibility to prostaglandin F2α (PGF2α) luteolysis in hysterectomized prepuberal and mature gilts

The susceptibility of induced corpora lutea (CL) of prepuberal gilts and spontaneously formed CL of mature gilts to prostaglandin F_2α (PGF_2α) luteolysis was studied. Prepuberal gilts (120 to 130 days of age) were induced to ovulate with Pregnant Mare Serum Gonadotropin and Human Chorionic Gonadotropin (HCG). The day following HCG was designated as Day 0. Mature gilts which had displayed two or more estrous cycles of 18 to 22 days were used (onset of estrus = Day 0). Gilts were laparotomized on Day 6 to 9, their CL marked with sterile charcoal and totally hysterectomized. On Day 20, gilts were injected IM with either distilled water (DW), 2.5 mg PGF_2α or 5.0 mg PGF_2α. An additional group of prepuberal gilts was injected with 1.25 mg PGF_2α, a dose of PGF_2α equivalent, on a per kilogram body weight basis, to the 2.5 mg PGF_2α dose given to the mature gilts. The percentages of luteal regression on Day 27 to 30 for mature and prepuberal gilts given DW, 2.5 mg PGF_2α and 5.0 mg PGF_2α were 0.0 vs 4.4, 43.5 vs 96.8 and 47.7 vs 91.6, respectively; the percentage of luteal regression for the prepuberal gilts given 1.25 mg PGF_2α was 75.1. These results indicate that induced CL of the prepuberal gilt were more susceptible to PGF_2α luteolysis than spontaneously formed CL of the mature gilt and that pregnancy failure in the prepuberal gilt could be due to increased susceptibility of induced CL to the natural luteolysin.     

Effect of intravenous infusion of 15-methyl-PGF2α on plasma hormone levels during early pregnancy

Ten pregnant women (7th–8th week of pregnancy) obtained an intravenous infusion of 15-methyl-prostaglandin-F_2α (2.5 μg/min) until clinical signs of abortion occurred or up to 7 hours. Surgical removal of the products of conception was performed 4–7 hours after the start of the infusion. Blood samples were taken prior to and during the infusion and then during the post-abortion period. The plasma levels of both progesterone and estradiol exhibited a significant decrease (p<0.001 and p<0.05, respectively) one hour after the beginning of infusion and remained reduced by approximately 35 and 45 per cent, respectively, during the entire infusion period. A drop in the levels of both steroids was seen after surgical interruption. This was followed by a gradual decrease to non-pregnancy levels. The levels of cortisol increased significantly (p<0.01) by approximately 60 per cent, starting with the second hour of infusion. Following surgical interruption, the levels dropped to pre-infusion values. 17-Hydroxyprogesterone levels increased (p<0.05) above the pretreatment levels by approximately 25 per cent, starting with the third hour of infusion. These levels were not correlated with those of cortisol during the infusion period. Following surgical interruption the plasma levels of 17-hydroxyprogesterone returned to non-pregnancy levels.     

Termination of pregnancy in cats with prostaglandin F2α

Two subcutaneous injections of Prostaglandin F_2α THAM salt 24 hours apart terminated pregnancies in cats after the 40th day of gestation. Injections of 0.50 or 1.00 mg. PGF_2α THAM salt/Kg. body weight were the most effective in terminating pregnancies. Parturition or abortion occurred within 24 hours after the initial injection in 9 cats and after the 2nd injection in 4 cats.