The influence of xanthines on the contractile responses of PGF2 alpha and PGD2 in human parenchymal strips

Prostaglandins (PGs) F2 alpha and D2 are bronchoconstrictor agents which are released under allergic conditions such as asthma. The efficacy and potency of PGF2 alpha and PGD2 differ in some tissues. We compared the effects of these two PGs in sensitized human parenchymal strips. In six experiments, PGF2 alpha 0.1 and 0.3 microM produced greater contractions than PGD2 at the same concentrations. There were no significant differences between the contractions from the two PGs at concentrations of 0.01, 0.03, 1.0-10 microM and the two PGs appeared to be equipotent. We studied the effects of the anti-asthmatic drug theophylline, and its analogue enprofylline, on the contraction caused by these PGs. Theophylline 100 microM caused no change to the cumulative concentration response curves. However, enprofylline 100 microM reduced the PGF2 alpha-induced contractions.     

A comparison of the contractile activity fo PGD2 and PGF2α on human isolated bronchus

Prostaglandins may be implicated in the bronchoconstriction which occurs in asthma. Prostaglandins F_2α (PGF_2α and D₂ (PGD₂) have been reported to produce bronchoconstriction in asthmatic subjects in vivo and PGF_2α cotnracts human isolated airway smooth muscle. We examined the relative efficacy and potency of PGF_2α and PGD₂ on human bronchial spiral strips taken from 6 patients at thoracotomy. PGF_2α had greater efficacy than PGD₂. The mean % T_max (percentage of maximal contractile response) ± s.e. mean were 84 ± 7 and 54 ±7 respectively (P < 0.05). PGF_2α (mean pD₂ ± s.e. mean = 6.39 ± 0.6) tended to be more potent than PGD₂ (5.68 ± 0.2). Since, in vivo, PGD₂ has greater efficacy and potency than PGF_2α, our results suggest that the in vivo effect of these prostaglandins does not result solely from an action on airway muscle.     

Evidence for separate PGD2 and PGF2 alpha receptors in the canine mesenteric vascular bed.

Potential interactions between PGD2 and PGF2 alpha in the mesenteric and renal vascular beds were investigated in the anesthetized dog. Regional blood flows were measured with electromagnetic flow probes. PGD2, PGF2 alpha and Norepinephrine (NE) were injected as a bolus directly into the appropriate artery, and responses to these agents were obtained before, during and after infusion of either PGD2 or PGF2 alpha into the left ventricle. In each case, the infused prostaglandin caused vascular effects of its own. Left ventricular infusion of PGD2 reduced responses to local injections of PGD2 in the intestine, and a similar effect was observed for PGF2 alpha, suggesting significant receptor or receptor-like interactions for each of the prostanoids. However, systemic infusion of prostaglandin F2 alpha (20--100 ng/kg/min) had no effect on renal or mesenteric vascular responses to local injection of prostaglandin D2. Similarly, PGD2 administration (100 ng/kg/min) did not affect responses to PGF2 alpha in the intestine. The present results therefore suggest that these prostaglandins, i.e., D2 and F2 alpha, act through separate receptors in the mesenteric and renal vascular beds. In addition, increased prostaglandin F2 alpha levels produced by infusion of F2 alpha reduced mesenteric but not renal blood flow, suggesting that redistribution of cardiac output might participate in side effects often observed with clinical use of this prostaglandin, such as nausea and abdominal pain.     

Serum PGF2alpha levels during human pregnancy

Guttierez Cernosek and his colleagues reported that the level of PGF2a in human serum reached a peak during the second trimester of pregnancy, the highest levels being found during the 17th-24th weeks. Earlier studies in this laboratory on late pregnancy have now been extended to earlier gestations and rather than peak levels during the 17th-24th weeks, the lowest levels were found at this time. A total of 128 samples from 106 women were studied. The levels found during the second trimester were significantly lower than those found at earlier gestations. However, the mean serum PGF2a level during the second trimester was not significantly different from the mean level during the third trimester. The discrepancy between the 2 sets of results is very marked, so that there is obviously need for much further research in this sphere.     

Influence of PGF2 alpha on gastrointestinal activity in the conscious piglet.

In 5 conscious piglets with electrodes implanted on the antrum pylori, duodenum, jejunum and ileum, the effect of intravenous infusion of PGF2 alpha, 1 and 10 micrograms/kg/min during 2 h, on gastrointestinal electrical activity was studied. The influence of the PG, 10(-8) to 10(-4) M, on longitudinal tissue strips from the same segments was also examined. The in vitro results demonstrate that PGF2 alpha has only a weak contractile effect on duodenal and jejunal strips. This effect was enhanced in the presence of atropine and indomethacin. In the in vivo part of the study PGF2 alpha induced an inhibition of antral electrical activity as evidenced by a prolongation of the inhibitory phases and a reduction of the frequency of the fast oscillations. In the small intestine only ileal activity was changed significantly. PGF2 alpha provoked an increase in the phase II or irregular spiking activity and an increase in the interval of the migrating myoelectrical complexes in this segment.     

Estrus synchronization in beef cows: comparison between GnRH+PGF2alpha+GnRH and PRID+PGF2alpha+eCG

The aim of this study was to compare two protocols for estrus synchronization in suckled beef cows over a 2 years period. The population studied consisted of 172 Charolais and 168 Limousin cows from 12 and 14 beef herds, respectively. In each herd, cows were allotted to groups according to parity, body condition score and calving difficulty. Cows in Group 1 (n=174) received PRID on Day-8 with estradiol benzoate (10mg, vaginal capsule), dinoprost on Day-4 (25mg i.m.), eCG on Day 2 (500 IU i.m.). The PRID was removed on Day-2 and cows were inseminated on Day 0, 56 h after PRID was removed. Cows in Group 2 (n=166) received GnRH on Day-10 (100 microg i.m.), dinoprost on Day-3 (25mg i.m.) and GnRH on Day-1 (100 microg i.m.), and were inseminated on Day 0, 16-24h after the last GnRH treatment. Plasma progesterone concentrations were measured to determine cyclicity prior to treatment (Days-20 and -10), to confirm the occurrence of ovulation (Days 0 and 10) and to determine the apparent early pregnancy rate (Days 0, 10 and 24). Pregnancy diagnosis was performed by ultrasonography between Days 35 and 45. The effects of various factors on ovulation, apparent early pregnancy and pregnancy rates were studied using logistic mixed models. There was no significant difference between Groups 1 and 2, respectively, for the cyclicity rate before treatment (80.5% versus 80.1%), for apparent pregnancy rate on Day 24 (62.1% versus 54.8%, P=0.09) and for pregnancy rate on Days 35-45 (53.8% versus 46.3%, P=0.16). Ovulation rate was higher (P<0.01) in Group 1 (90.8%) than in Group 2 (77.1%) and was affected by cyclicity prior to treatment in Group 2 but not in Group 1 (Group 1: 88.2% in anestrous cows versus 91.4% in cyclic cows; Group 2: 45.5% in anestrous cows versus 85.0% in cyclic cows, P interaction=0.05). Apparent pregnancy rates on Day 24 were influenced by the year of study (52.4% versus 68.8%, OR=2.12, P<0.01) and by the cyclicity before treatment (anestrous cows 46.3% versus cyclic cows 61.5%, OR=1.86, P<0.05). Pregnancy rates at 35-45 days were influenced by the year of study (44.2% versus 59.8%, OR=1.92, P<0.01). In conclusion, although pregnancy rates were similar for the two treatments, the combination of GnRH+PGF2alpha+GnRH in suckled beef cows induced a lower rate of ovulation than treatment with PRID+PGF2alpha, particularly in anestrous cows.     

A sheer pharmacologic approach to compare the contractile effects of PGF2alpha, DL-cloprostenol and D-cloprostenol on isolated uterine, tracheal, ileal and arterial smooth muscle preparations

The contractile effects of PGF2alpha and its cloprostenol analogs (D-enantiomer and racemate) were examined on isolated smooth muscle preparations (uterine, tracheal, ileal and arterial) from rat, guinea-pig and horse. DL- and D-cloprostenol were potent contractors of myometrium, but had negligible secondary effects on other types of smooth muscle, except artery whose response to the D-enantiomer may give rise to some concern.     

Proliferative effect of PGD2 on osteoblast-like cells; independent activation of pertussis toxin-sensitive GTP-binding protein from PGE2 or PGF2 alpha.

PGD2 stimulated DNA synthesis and decreased alkaline phosphatase activity dose-dependently between 10 nM and 10 microM in osteoblast-like MC3T3-E1 cells. PGD2 had little effect on cAMP production, but caused very rapid enhancement of phosphoinositide (PI) hydrolysis dose-dependently between 10 nM and 10 microM. The formation of inositol trisphosphate (IP3) induced by PGD2 reached the peak within 1 min and decreased thereafter, which is more rapid than that induced by PGE2 or PGF2 alpha and both PGE2 and PGF2 alpha affected PGD2-induced IP3 formation additively. Pertussis toxin (PTX) inhibited both PGD2-induced formation of inositol phosphates and DNA synthesis. The degree of these PTX (1 micrograms/ml)-induced inhibitions was similar. In addition, neomycin, a phospholipase C inhibitor, inhibited PGD2-induced DNA synthesis as well as the formation of IP3, and the patterns of both inhibitions were similar. In the cell membranes, PTX-catalyzed ADP-ribosylation of a 40-kDa protein was significantly attenuated by pretreatment of PGD2. Time course of the attenuation of PTX-catalyzed ADP-ribosylation by PGD2 was apparently different from that by PGE2 or PGF2 alpha. These results indicate that PGD2 activates PTX-sensitive GTP-binding protein independently from PGE2 or PGF2 alpha and stimulates PI hydrolysis resulting in proliferation of osteoblast-like cells.     

Luteolytic activity of a synthetic prostaglandin and PGF2alpha in heifers.

Two experiments involving 44 cycling heifers were conducted to evaluate the luteolytic activity of a synthetic prostaglandin, AY 24366, and PGF2alpha. Activity was assessed by the decline in progesterone level of peripheral blood and occurrence of estrus. Progesterone concentrations of jugular blood plasma were quantified by radioimmunoassay. In the first experiment, 36 heifers were treated during diestrus with AY 24366 (A-10mg intrauterine, B-30mg intramuscular and C-60mg im) or with PGF2alpha (D-5mg iu, E-15mg im and F-30mg im). Mean progesterone 0, 24 and 48 hours after treatment were A-6.33, 5.55 and 5.06; B-6.35, 2.79 and 3.92; C-5.23, 2.69 and 3.91; D-5.19, 1.50 and 1.51; E-4.69, 0.85 and 0.61; F-6.66, 0.80 and 0.48 ng/ml. Standing estrus was observed in 1, 1, 1, 4, 5 and 6 females in groups A, B, C, D, E and F respectively within 72 hours of treatment. PGF2alpha resulted in significantly (P less than 0.01) lower progesterone at 24 and 48 hours than AY 24366. However, in administration of the latter did significantly (P less than 0.05) lower progesterone at 24 hours. In the second trial six heifers were treated with either 120 or 180mg of AY 24366 im on day 12 of the cycle. Mean progesterone declined from 3.84 to 2.12 ng/ml (P less than 0.01) by 6 hours and to 1.59 ng/ml by 12 hours. Thereafter the decline was gradual and reached a level of 0.65 ng/ml at 72 hours. All six heifers showed standing estrus at 78 +/-2 hours and were inseminated. Two in each group conceived. Doses of 15mg PGF2alpha and 120mg AY 24366 were effective in causing luteal regression, however, the latter caused respiratory discomfort for 5 to 10 minutes post treatment.     

Cyclic AMP formation of isolated human corpora lutea in response to HCG - interference by PGF2 alpha.

Human corpora lutea of defined ages were excised at operation, cut into pieces and incubated in the presence of HCG, PGF2 alpha and PGE2 alone or in combination. Following incubation cAMP formation in tissue and medium was determined. HCG-stimulated tissue cAMP content was most pronounced at a corpus luteum age of 7-10 days after ovulation. This stimulation was antagonized by PGF2 alpha in corpora lutea older than 6 days. PGE2 stimulated cAMP formation per se and this effect was more pronounced when HCG and PGE2 were combined. A possible role for PGF2 alpha as a luteolytic substance in the human is suggested.     

Radioimmunoassay measurement of ACTH-facilitated PGE2 and PGF2alpha release from isolated cat adrenocortical cells.

Prostaglandin (PG) biosynthesis by trypsin-dispersed cat adrenocortical cells was studied by radioimmunoassay (RIA). Parallel assays of incubation media using PGF2alpha and PGF1alpha antisera established that PGF2alpha is the primary PGF released by feline cortical cells. Following the reduction of PGE to PGF with sodium borohydride (NaBH4) these same two antisera were also used to identify PGE2 as the primary PGE released. RIA using a PGE antiserum confirmed the presence of PGE in the incubation medium. Steroidogenic concentrations of ACTH (50-250muU) enhanced PGE and PGF release, and indomethacin suppressed the ACTH-facilitated release. These studies provide additional evidence for ACTH-induced PG synthesis by feline cortical cells, and support the hypothesis that PGs play some role in the steroidogenic action of ACTH.     

Effects of PGF2alpha on human melanocytes and regulation of the FP receptor by ultraviolet radiation

Prostaglandins are potent lipid hormones that activate multiple signaling pathways resulting in regulation of cellular growth, differentiation, and apoptosis. In the skin, prostaglandins are rapidly released by keratinocytes following ultraviolet radiation and are chronically present in inflammatory skin lesions. We have shown previously that melanocytes, which provide photoprotection to keratinocytes through the production of melanin, express several receptors for prostaglandins, including the PGE2 receptors EP1 and EP3 and the PGF2alpha receptor FP, and that PGF2alpha stimulates melanocyte dendricity. We now show that PGF2alpha stimulates the activity and expression of tyrosinase, the rate-limiting enzyme in melanin synthesis. Analysis of FP receptor regulation showed that the FP receptor is regulated by ultraviolet radiation in melanocytes in vitro and in human skin in vivo. We also show that ultraviolet irradiation stimulates production of PGF2alpha by melanocytes. These results show that PGF2alpha binding to the FP receptor activates signals that stimulate a differentiated phenotype (dendricity and pigmentation) in melanocytes. The regulation of the FP receptor and the stimulation of production of PGF2alpha in melanocytes in response to ultraviolet radiation suggest that PGF2alpha could act as an autocrine factor for melanocyte differentiation.     

Effects of PGF2alpha and PGF2alpha, 1-15 lactone on the corpus luteum and on early pregnancy in the rhesus monkey.

The effects of prostaglandin (PG)F2alpha and PGF2alpha, 1-15 lactone were compared in luteal phase, non-pregnant and in early pregnant rhesus monkeys. Animals treated with either PG after pretreatment with human chorionic gonadotropin (hCG) had peripheral plasma progesterone concentrations that were not statistically different from those in animals treated with hCG and vehicle. However, menstrual cycle lengths in monkeys treated with PGF2alpha, 1-15 lactone were significantly (P less than 0.02) shorter than those in vehicle treated animals. In the absence of hCG pretreatment, plasma progesterone concentrations were significantly (P less than 0.008) lower by the second day after the initial treatment with either PGF2alpha or PGF2alpha, 1-15 lactone than in vehicle treated monkeys. Menstrual cycle lengths in monkeys treated with either PG were significantly (P less than 0.04) shorter than those in animals treated with vehicle. There were no changes in plasma progesterone concentrations in early pregnant monkeys treated with PGF2alpha, and pregnancy was not interrupted. In contrast, plasma progesterone declined and pregnancy was terminated in 5 of 6 early pregnant monkeys treated with PGF2alpha, 1-15 lactone. These data indicate that PGF2alpha, 1-15 lactone decreases menstrual cycle lengths in non-pregnant rhesus monkeys. More importantly, PGF2alpha, 1-15 lactone terminates early pregnancy in the monkey at a dose which is less than an ineffective dose of PGF2alpha.     

Spontaneous contractile activity of isolated ovarian human vein. A dual influence of prostacyclin (PGI2).

The profile of spontaneous contractions as well as the influences of prostacyclin (PGI2) on the motility of human ovarian veins obtained during the estrogenic phase of the sex cycle were explored. The preparations exhibited a distinct phasic activity, which progressively decreased as isolation time progresses, dissapearing almost completely following more than two hours. PGI2 produced a biphasic influence on quiescent preparations. After the threshold is attained lower concentrations caused depression of tone whereas higher ones enhanced the basal tone and induced phasic contractile cycles. Phentolamine reduced markedly the stimulating influence of PGI2 but had no action on the inhibitory effects, whereas propranolol failed to alter either the excitatory of the depressive action. The results suggest a participation of alpha-adrenoceptive-mediated mechanisms in the stimulatory effect of PGI2. On the other hand, PGI2 may be of importance in the regulation of venous flow and the spontaneous or PGI2-induced contractions could play a role in the counter current mechanism between veins and arteries in the ovarian pedicle.     

Self-differentiation of human fetal lung organ culture: the role of prostaglandins PGE2 and PGF2 alpha

Addition of PGE2, but not PGF2 alpha, to fetal lung organ cultures accelerates the process of self-differentiation with increased dilatation of terminal airsacs and differentiation of the epithelial lining. Indomethacin reduces the endogenous production by organ cultures of PGE2, PGF2 alpha, 13,14-dihydro-15-keto-PGE2, and 13,14-dihydro-15-keto-PGF2 alpha and retards the process of self-differentiation. Prolonged exposure of cultures to indomethacin results in cell necrosis. Indomethacin inhibition of self-differentiation can be reversed and accelerated by the addition of PGE2. Addition of PGF2 alpha in the presence of indomethacin prevents indomethacin-associated cell necrosis but does not accelerate dilatation or differentiation beyond that of cultures in sera-free media without additions. We propose that the endogenous production of PGE2 is a key process in the mechanism of self-differentiation of human fetal lung in organ culture.