Search PubMed⌕ Search

Biomedical subjects

L L Espey

Publications and source records attributed to L L Espey.

At least 19 recordsLinked to original sources
PubMed1992

Inhibition of ovulation in the gonadotropin-primed immature rat by exogenous prostaglandin E2.

In the past two decades there have been innumerable reports that prostaglandins (PGs) are essential for mammalian ovulation. However, we have recently found that a relatively low dose of 0.03 mg indomethacin (INDO) sc to PMSG/hCG-primed immature Wistar rats can significantly reduce ovarian PG levels without inhibiting the control ovulation rate of 60+ ova/rat (1-3). In view of this information, the present study was an effort to duplicate the earlier reports that PGs can reverse the "inhibitory" effect of INDO on ovulation. In control animals, which received PMSG and hCG only, the ovulation rate was 63.8 +/- 4.5 ova/rat. This rate was reduced to 4.1 +/- 1.1 ova/rat when the animals were injected with 1.0 mg INDO at 3 h after hCG. In no instance was this inhibition reversed when the animals were treated with 1.0 mg of PGE2 or PGF2 alpha, or a combination of both prostanoids in either a single dose at 3 h after hCG, or in 4x doses at 2-h intervals beginning at 3 h after hCG. Furthermore, in animals that did not receive INDO, the ovulation rate in PGE2-treated animals was reduced to 20.0 +/- 6.7 ova/rat, and in animals treated with PGE2 and PGF2 alpha (combined) it was reduced to 19.4 +/- 6.5 ova/rat. In summary, not only did the PGs fail to reverse the anti-ovulatory effect of INDO, PGE2 actually suppressed the ovulation rate.

Animals↗
PubMed1992

Epostane and indomethacin actions on ovarian kallikrein and plasminogen activator activities during ovulation in the gonadotropin-primed immature rat.

Kallikrein and plasminogen activator (PA) are serine proteases that have been implicated in the ovulatory process. Epostane and indomethacin are anti-ovulatory agents that inhibit steroid and eicosanoid synthesis, respectively. This study examines the effects of these two anti-ovulatory agents on ovarian kallikrein and PA activities during ovulation. The proteases were assayed by their actions on chromogenic peptide substrates S-2266 and S-2251, respectively. The ovulatory process was induced in 25-day-old Wistar rats by giving them hCG (10 IU, s.c.) 2 days after the animals had been primed with eCG (10 IU, s.c.). Control animals ovulated approximately 60-70 ova/rat, with the first ova appearing in the oviducts at 10-12 h after hCG administration, and this was the same time ovarian kallikrein and PA activities reached a peak. When doses of epostane ranging from 0.1-5.0 mg/rat or doses of indomethacin ranging from 0.03 to 3.16 mg/rat were administered s.c. at 3 h after hCG, the two drugs inhibited ovulation and ovarian kallikrein and PA activities in a dose-dependent manner. However, the anti-ovulatory action of the two drugs was more closely correlated with suppression of kallikrein activity than with PA activity. Treatment of the animals with exogenous progesterone reversed the inhibitory action of epostane, but not of indomethacin. The results suggest that the increase in ovarian progesterone at the time of ovulation may influence ovarian kallikrein and PA activities.

Age Factors↗
PubMed1991

Ovarian hydroxyeicosatetraenoic acids compared with prostanoids and steroids during ovulation in rats.

Hydroxyeicosatetraenoic acid methyl esters (HETEs) are lipoxygenase products of arachidonic acid that are generated along with prostaglandins (PGs) during acute inflammatory reactions. Whereas it is well known that ovarian PG levels increase during the ovulatory process, little is known about ovarian HETEs. This report compares the ovarian changes in 5-, 12-, and 15-HETE with ovarian PGE and PGF, along with progesterone, 17 alpha-hydroxyprogesterone, 4-androstene-3,17-dione, testosterone, and 17 beta-estradiol. Ovulation was induced in immature Wistar rats by sequential treatment with pregnant mare's serum gonadotropin and human chorionic gonadotropin (hCG). Follicles began rupturing 10 h after hCG treatment. The greatest correlation was among 12-HETE, 15-HETE, and progesterone, which increased to peak levels at 10 h after hCG. In contrast, the ovarian levels of 5-HETE, 17 alpha-hydroxyprogesterone, testosterone, and 17 beta-estradiol all declined sharply beginning 4 h after hCG. 2 alpha,4 alpha,7-4,5-Epoxy-17-hydroxy-4,17-dimethyl-3-oxo-androstane-2- carbonitrile (epostane), a potent inhibitor of steroid synthesis and ovulation, sharply reduced the synthesis of all five steroids within 30 min after its injection at 3 h after hCG. Among the five eicosanoids, epostane mainly inhibited 15-HETE. The results suggest that 15-HETE, along with progesterone, may have an important role in ovulation.

17-alpha-Hydroxyprogesterone↗
PubMed1991

Comparison of inhibitory actions of indomethacin and epostane on ovulation in rats.

Indomethacin, an inhibitor of cyclooxygenase that generates prostaglandins (PGs) from arachidonic acid, and 2 alpha,4 alpha,7-4,5-epoxy-17-hydroxy-4,17-dimethyl-3-oxoandrostane- 2-carbonitrile (epostane), an inhibitor of 3 beta-hydroxysteroid dehydrogenase that generates progesterone from pregnenolone, are both potent inhibitors of ovulation. This report compares the dose-dependent effects of these two inhibitors on ovarian levels of 5-, 12-, and 15-hydroxyeicosatetraenoic acid methyl ester (HETEs), prostaglandin E2 (PGE), prostaglandin F2 alpha (PGF), progesterone, 17 alpha-hydroxyprogesterone, 17 beta-estradiol, 4-androstene-3,17-dione, and testosterone during ovulation in 25-day-old immature Wistar rats. The ovulatory process was initiated by 10 IU of human chorionic gonadotropin (hCG). Indomethacin was given at 3 h after hCG in doses ranging from 0.0316 to 10.0 mg/rat. A dose of 0.1 mg/rat was the lowest dose to significantly reduce the ovulation rate from the control level of 70.5 +/- 5.8 ova/rat. This dose also reduced 15-HETE, but not 5-HETE, 12-HETE, or the steroids. PGE and PGF were strongly inhibited by an even lower dose of indomethacin (0.0316 mg/rat), but this dose did not affect the ovulation rate. Epostane was given at 3 h after hCG in doses ranging from 0.1 to 5.0 mg/rat. A dose of 1.0 mg/rat was the lowest dose to significantly inhibit ovulation. This dose also reduced the ovarian levels of 15-HETE and progesterone but not 5-HETE, 12-HETE, PGE, PGF, or the other steroids. The results indicate that the ovulation rate is most closely correlated to ovarian 15-HETE levels.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1990

Effects of epostane on ovarian levels of progesterone, 17 beta-estradiol, prostaglandin E2, and prostaglandin F2 alpha during ovulation in the gonadotropin-primed immature rat.

The antiovulatory action of epostane, an inhibitor of 3 beta-hydroxysteroid dehydrogenase activity and progesterone synthesis, was studied in the immature rat. The ovulatory process was induced in 25-day-old rats by injecting them with hCG (10 IU, sc) 2 days after the animals had been primed with PMSG (10 IU). Epostane was administered at different times between 20 h before and 11 h after hCG. Maximum inhibition of ovulation occurred when the drug was given at 3 h after hCG. Epostane inhibited ovulation in a dose-dependent manner when administered in doses ranging from 1.0-50 mg/rat, while exogenous doses of progesterone restored the ovulation rate. A dose of 3.1 mg epostane/rat 3 h after hCG reduced ovarian progesterone levels within 15 min, but the production of this steroid rebounded within 2 h and approached normal levels by 12 h after hCG, i.e. when the follicles began to rupture in control animals. 17 beta-Estradiol synthesis was inhibited just as rapidly, but it remained suppressed for up to 12 h after hCG administration. The ovarian levels of prostaglandins E2 and F2 alpha decreased approximately 30% within 2 h after the administration of epostane, but such a moderate reduction in the synthesis of ovarian prostanoids is usually not sufficient to block ovulation. The results show that epostane has a rapid, but transient, effect on ovarian progesterone synthesis. The temporary decline in the local progesterone level is apparently sufficient to interfere with the normal sequence of metabolic events that lead to the rupture of follicles.

3-Hydroxysteroid Dehydrogenases↗
PubMed1990

Characteristics and control of the normal menstrual cycle.

The most important activity during the follicular phase of the cycle is the secretion of gonadotropins, which control folliculogenesis and influence uterine proliferation. The dominant events of the periovulatory phase are the LH surge and ovulation. The significant change during the luteal phase is the production of a nutritive mucus by the endometrial glands in preparation for an embryonic blastocyst. The cardinal passage of the menstrual phase is the menstrual flow itself. These different events (and the metabolic processes that regulate them) have wide-ranging effects on the integrity of the body. As Havelock Ellis, the eminent English psychologist, stated at the turn of this century, "the omnipresent process of sex, as it is woven into the whole texture of our body, is the pattern of all the process of our life" (source unknown). The sweeping influences of the menstrual cycle illustrate the extent to which the process of reproduction is indeed woven into the whole of the human body.

Female↗
PubMed1989

Effect of indomethacin, cycloheximide, and aminoglutethimide on ovarian steroid and prostanoid levels during ovulation in the gonadotropin-primed immature rat.

It has become popular to use the gonadotropin-primed immature rat to study ovulation. The ovarian content of progesterone, estradiol, PGE2, PGF2 alpha, and 6-keto-PGF1 alpha during the ovulatory process was determined in this model. Also, the effect of three anti-ovulatory agents on the ovarian levels of the above substances was determined. At 23 days of age, Wistar rats were primed with pregnant mares serum gonadotropin (PMSG) sc, and two days later the ovulatory process was initiated with human chorionic gonadotropin (hCG) sc. The ovarian follicles began rupturing 12 h later. Ovaries were assayed for the two steroids and prostanoids at 2-h intervals before and several 4-h intervals after ovulation. The ovarian estradiol level increased slightly between 0 and 2 h after hCG, while the progesterone level increased sharply between 2 and 4 h after hCG--at a time when the estradiol declined markedly. All three prostanoids increased concomitantly with progesterone. When the PG synthesis was blocked by indomethacin treatment at 1 h before hCG, ovarian progesterone levels still increased. In contrast, when steroidogenic activity was inhibited by aminoglutethimide, the ovarian prostanoid levels also decreased. Cycloheximide had little effect on the steroids and prostanoids. It is concluded that ovarian prostanoid synthesis might be influenced by ovarian steroid output.

Aminoglutethimide↗
PubMed1989

Increase in ovarian blood volume during ovulation in the gonadotropin-primed immature rat.

This study quantifies ovarian blood volume in Wistar rats by measuring the optical density (414 nm) of hemoglobin in ovarian extracts and comparing this measurement to the optical density of known amounts of whole blood. Immature rats were primed with pregnant mare's serum gonadotropin (PMSG), 10 IU s.c., at 23 days of age. On Day 25, the ovulatory process was initiated by human chorionic gonadotropin (hCG), 10 IU s.c., and ova began to appear in the oviducts 10 h later. At 2-h intervals, the ovaries were extirpated and homogenized in 1.0 ml of 0.05 M tris (hydroxymethyl)aminomethane buffer (pH 7.4) for 30 s. Homogenates were centrifuged for 20 min and the supernatant fluids were analyzed with a Gilford RESPONSE UV/VIS spectrophotometer. The hemoglobin in these ovarian extracts had the same peak absorbance of 414 nm characteristic of oxyhemoglobin in whole blood taken by cardiac puncture of the rats. There was a linear relationship between the absorbance and the volume of whole blood in the samples. The volume of blood per ovary from groups of 8 rats was 0.60 +/- 0.07 microL at 0 h after hCG. The volume increased to 1.37 +/- 0.26 microL at 4 h after hCG and reached a peak of 4.55 +/- 0.72 microL at 10 h. Indomethacin treatment (0.3-10.0 mg/rat, s.c.) partially inhibited this 7-fold increase in ovarian blood volume. In conclusion, the increase in ovarian blood volume during ovulation may reflect the vasodilation and hyperemia that are characteristic of inflamed tissues.

Animals↗
PubMed1989

Decrease in ovarian platelet-activating factor during ovulation in the gonadotropin-primed immature rat.

Platelet-activating factor (PAF) is a biologically active phospholipid that is released locally during acute inflammatory reactions and tissue injury. Since there is evidence that the biochemical events of mammalian ovulation resemble an inflammatory reaction, the objective of this study was to determine whether ovarian levels of PAF change during ovulation. At 2-h intervals during the ovulatory process in gonadotropin-primed 25-day-old Wistar rats, the ovaries were extirpated, homogenized, and extracted for lipids. The extracts were subjected to thin-layer chromatography (TLC), and the portion of the silica gel that comigrated with PAF was re-extracted and assayed for PAF activity. The PAF was measured (in fmole equivalents of synthetic PAF) by a bioassay based on the capacity of aliquots of the extracts to release [3H]-serotonin from platelets isolated from whole blood of rabbits and prelabeled with [3H]-serotonin. The ovarian level of PAF decreased (p less than 0.01) by 36% from 6.67 +/- 0.77 to 4.27 +/- 0.45 fmoles/mg ovary by 2 h after treatment with human chorionic gonadotropin (hCG), and it declined another 14% by 4 h after hCG. The ovarian PAF remained at this reduced level for up to 24 h after hCG. The administration of indomethacin (5 mg/rat, s.c.) or epostane (5 mg/rat, s.c.) at 1 h after hCG prevented ovulation, but neither drug affected the decline in ovarian PAF. Preliminary tests showed that the lipid extracts from the ovaries also contained PAF inhibitor(s) that comigrated with PAF on the TLC plates. Similar to PAF, the lipid-soluble inhibitor(s) decreased (p less than 0.05) in the ovaries within 4 h after hCG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Androstenols↗
PubMed1989

Increase in ovarian leukotrienes during hormonally induced ovulation in the rat.

The ovulatory process was initiated in 25-day-old Wistar rats by injecting human chorionic gonadotropin (hCG; 10 IU sc) 2 days after the animals had been primed with pregnant mares serum gonadotropin (PMSG; 10 IU sc). By 4 h into the ovulatory process, leukotriene (LT) B4 increased 2-fold (P less than 0.001) and LTs C4/D4/E4 increased 1.3-fold (P less than 0.002). By the time of ovulation (10-12 h after the administration of hCG) both eicosanoids declined to their pre-hCG levels. When animals were treated with the cyclooxygenase inhibitor indomethacin at the specific dose of 0.316 mg/rat sc at 1 h before hCG, the ovarian levels of LT B4 and LTs C4/D4/E4 increased to 210% (P less than 0.01) and 113% (P less than 0.05), respectively, above the control levels at 4 h after hCG. Concomitantly, this dosage of indomethacin reduced ovarian prostaglandins (PGs) E and F by 99% (P less than 0.001) and 98% (P less than 0.001), respectively, and it reduced the ovulation rate by 76% (P less than 0.001). Thus it appears this dose of indomethacin blocked the conversion of ovarian arachidonic acid into PGs and shunted this substrate into the lipoxygenase pathways that lead to LT formation. In conclusion, the moderate increase in ovarian LTs is characteristic of inflammatory reactions, and, therefore, these data support the hypothesis that the biochemical events of ovulation resemble an inflammatory process.

Animals↗
PubMed1989

Increase in ovarian 15-hydroxyeicosatetraenoic acid during ovulation in the gonadotropin-primed immature rat.

The ovarian level of 15-hydroxyeicosatetraenoic acid (15-HETE) was measured by RIA during ovulation in gonadotropin-primed immature Wistar rats. The ovulatory process was initiated in 25-day-old rats by a 10-IU injection of hCG sc 2 days after the animals had been primed with 10 IU PMSG, sc. Ovarian follicles begin to ovulate 10 h after hCG. At 0 h after hCG, the ovarian 15-HETE level was 0.6 +/- 0.2 ng/mg ovarian protein. At 6 h the 15-HETE level increased sharply to 27.3 +/- 4.2 ng/mg protein and reached a peak of 50.0 +/- 9.8 ng/mg protein at 10 h. Ovarian 15-HETE decreased significantly between 10-16 h after hCG (when ovulation was essentially completed). The pattern of secretion of this 15-lipoxygenase product was reciprocal to the pattern of secretion of leukotriene-B4 by the rat ovary. Ovarian 15-HETE production and ovulation were inhibited in a dose-dependent manner when indomethacin was administered sc 1 h after hCG in doses ranging from 0.10-10.0 mg/rat. In contrast, the synthesis of ovarian prostaglandins-E and -F was inhibited by a dose of indomethacin as low as 0.0316 mg/rat, but this dose did not significantly affect the ovarian 15-HETE level or the ovulation rate. Therefore, the ovulation rate was more closely correlated with the ovarian 15-HETE level (P less than 0.001) than with the ovarian levels of either prostaglandin-E or -F (0.10 greater than P greater than 0.05). The results suggest that products of the 15-lipoxygenase pathway of arachidonic acid metabolism may be important in the biochemical events of mammalian ovulation.

Animals↗
PubMed1989

Increase in ovarian kallikrein activity during ovulation in the gonadotrophin-primed immature rat.

The ovulatory process was initiated in 25-day-old rats by injecting them with hCG (10 i.u., s.c.) 2 days after the animals had been primed with PMSG (10 i.u., s.c.). At 2-h intervals after hCG, the ovaries were extracted and assayed for glandular kallikrein activity by using a chromogenic substrate (H-D-Val-Leu-Arg-p-nitroanilide) which exhibits optical density (at 405 nm) upon hydrolysis. In 0-h control ovaries the activity was 12.5 x 10(-3) kallikrein units (KU)/mg protein and it increased to a peak of 56.6 x 10(-3) KU/mg at 12 h after hCG, when the follicles first began to rupture. The kallikrein activity was distinguishable from ovarian plasminogen activator activity on the basis of pH optima and response to trypsin inhibitor (SBTI). The activity was inhibited by a s.c. dose of indomethacin of 0.3 mg/rat, or higher, and this dosage inhibited ovulation. The results suggest that kallikrein activity contributes to the degradation of Graafian follicles during ovulation in mammals.

Animals↗
PubMed1989

Pattern of ovarian steroid secretion during ovulation of in vitro perfused rat ovaries varies with method of sampling.

Wistar rats were primed at 25 days of age with 20 i.u. PMSG s.c., followed 48 h later with 100 ng LH/ml to induce the ovulatory process in vitro. Assays for progesterone, oestradiol, and testosterone were performed on samples taken at fixed intervals of 0, 1, 2, 4, 8 and 12 h after LH administration. The results show that, while there was some degree of correlation in the concentrations of progesterone between the ovarian extract and perfusate, there were very large differences in the measurable amounts of testosterone and oestrogen in the fluid samples and ovarian homogenates. Therefore, taking samples from the perfusate may not provide a good assessment of the steroidogenic activity in the ovary.

Animals↗
PubMed1988

Rat ovarian prostaglandin levels and ovulation as indicators of the strength of non-steroidal anti-inflammatory drugs.

Immature Wistar rats were treated with pregnant mare's serum gonadotropin and human chorionic gonadotropin to induce ovulation. The non-steroidal anti-inflammatory drugs indomethacin, diclofenac, flurbiprofen, and phenylbutazone inhibited both the ovulation rate and the normal increase in ovarian prostaglandin E during ovulation. Tolmetin, ibuprofen, and aspirin did not have any significant effect. There was a significant correlation between the ovulation rate and the level of ovarian prostaglandin E following treatment with these drugs. When indomethacin was given in graded doses, there was also a correlation between ovulation rate and the dose-dependent inhibition of ovarian prostaglandin E.

Animals↗
PubMed1986

Ovarian increase in kinin-generating capacity in PMSG/hCG-primed immature rat.

Ovarian kinin-generating capacity was determined during induced ovulation in immature Wistar rats. The onset of ovulation was monitored by counting the number of ova in the oviducts at 2-h intervals after the administration of human chorionic gonadotropin (hCG). Ova began to appear in significant numbers at 14 h after hCG, with an average of 7.6 +/- 2.3 ova/rat. By 16 h after hCG, the oviducts contained 32.7 +/- 4.1 ova/rat. The ovaries from each group of animals were homogenized in phosphate-buffered saline, and extracts of this tissue were incubated for 200 min to allow the generation of kinins from endogenous kininogen. The amount of kinin generated by this procedure was measured by radioimmunoassay. At 0 h (i.e., just before the administration of hCG), the ovaries contained 5.90 +/- 0.60 pg kinin/micrograms protein per 200 min in the ovarian extract. By 4 h after hCG, the kinins increased significantly (P less than 0.05) to 13.16 +/- 3.61 pg kinin/micrograms protein. The kinins progressively increased (P less than 0.001) to 67.88 +/- 23.26 pg kinin/micrograms protein by 16 h after hCG. Indomethacin and cycloheximide significantly inhibited both kinin-generating activity and ovulation. These data suggest that kinin-forming activity and kinins may have a role in the ovulatory process of mammals.

Animals↗
PubMed1986

Effect of time and dose of indomethacin on follicular prostaglandins and ovulation in the rabbit.

This study determined specifically when ovarian prostaglandins (PGs) increase during ovulation and how effectively different doses of indomethacin inhibit PGs and ovulation. Rabbit ovarian follicles were removed at hourly intervals after stimulating the animals with hCG (50 IU/kg). The follicles were homogenized in 0.1 M acetate buffer (pH 4.5), and the PGE and PGF in the extracts were measured by RIA. Ovulation rates were determined by calculating the percentage of mature follicles that ruptured after stimulation by hCG. Before hCG, the normal levels of PGE and PGF were 111.1 +/- 14.7 and 51.0 +/- 6.6 pg/mg follicle, respectively. By 2 h after hCG treatment, PGE and PGF both increased to 162.6 +/- 17.0 and 80.6 +/- 13.3 pg/mg follicle, respectively. Approximately 5 h later, there was a second, sharper increase in both PGs which peaked at 652.6 +/- 63.5 and 345.4 +/- 32.3 pg/mg follicle, respectively, 10 h after hCG treatment, i.e. at the expected time of ovulation. We found that regardless of whether indomethacin was given early or late during the ovulation process, this agent significantly reduced follicular PG production within 5 min after its administration. For example, only 5 min after 10 mg/kg indomethacin were administered 8 h after hCG, PGE and PGF dropped from 317.7 +/- 50.0 and 125.0 +/- 10.5 pg/mg follicle to 93.3 +/- 17.4 and 49.3 +/- 10.5 pg/mg follicle, respectively. Unexpectedly, when graded doses of indomethacin were administered either 1 or 8 h after hCG, there was not a statistically significant correlation between follicular PG levels and ovulation rate. For example, when doses of 1.25, 2.5, 5.0, and 10.0 mg/kg indomethacin were given 8 h after hCG, the PGE and PGF levels at the expected time of ovulation (i.e. 2 h later) were always equal to or less than the PG levels in follicles that had not been stimulated by hCG, yet ovulation proceeded at rates of 46.7 +/- 12.3%, 30 +/- 7.7%, 14.3 +/- 9.3%, and 0%, respectively. Therefore, the results raise questions about the specific role of PGs in the ovulation process.

Animals↗
PubMed1986

Cycloheximide inhibition of ovulation, prostaglandin biosynthesis and steroidogenesis in rabbit ovarian follicles.

Cycloheximide (5 mg/kg, i.v.) significantly inhibited ovulation in the rabbit when it was administered as early as 20 h before the ovulation process was initiated by hCG, and as late as 1 h after hCG. The ovulation rate was significantly reduced, but follicular biosynthesis of prostaglandins E and F was only partly inhibited. The biosynthesis of progesterone and oestradiol in follicles during the early stages of the ovulation process was also inhibited. Cycloheximide may therefore inhibit ovulation by a mechanism which is different from the action of indomethacin, and this mechanism may involve the suppression of ovarian steroidogenesis.

Animals↗