Controlled ovulation in the marmoset monkey (Callithrix jacchus) with human chorionic gonadotropin following prostaglandin-induced luteal regression

Controlled induction of ovulation in the marmoset monkey was attempted with a single injection of human chorionic gonadotropin (hCG; 50 IU) given on day 7 after prostaglandin-induced luteal regression. Animals given hCG (n = 12) ovulated within a 2-day period (days 9 and 10 after prostaglandin) compared with a 4-day period (days 9 to 12) in the control group (n = 12). The mean interval to ovulation was similar in both groups. There was no difference in the timing of the preovulatory estradiol (E2) peak between groups, although E2 levels on the day of hCG injection were lower than in controls on the day of the onset of the luteinizing hormone surge. All animals given hCG ovulated and 11 of 12 became pregnant. Ten of 11 embryos recovered surgically from six of these animals were normal blastocysts; 5 of the remaining 6 animals carried pregnancies to term. The results are of practical importance for experiments involving follicular and oocyte maturation and the collection and transfer of embryos.     

An extended regimen of clomiphene citrate in women unresponsive to standard therapy

An extended regimen of clomiphene consisting of 250 mg of clomiphene for 8 days followed by the administration of 10,000 IU of human chorionic gonadotropin (hCG) 6 days later was administered to 13 oligomenorrheic women who had previously failed to ovulate when treated with 250 mg of clomiphene for 5 days and hCG. Eight of these 13 women ovulated. Their postovulatory mean progesterone (P) level 7 days after hCG was 16 +/- 2 ng/ml. Three pregnancies occurred during 25 treatment cycles. Posttreatment estrogen levels were higher when women were treated for 8 days than for 5 days. Women ovulating after 8 days of treatment had increased concentrations of luteinizing hormone (LH) and testosterone (T) prior to hCG administration and higher pretreatment levels of estrogen and T, compared with women who did not ovulate. Changes in the timing of hCG administration may induce ovulation in some women who fail to ovulate when hCG is given on day 14. Because this 8-day regimen of clomiphene and hCG was successful in more than 50% of women failing to ovulate after 5 days, this regimen should be used prior to human menopausal gonadotropin (hMG) therapy.     

The effects of cisplatin on murine metaphase II oocytes.

This study was undertaken to determine if cisplatin can induce aneuploidy in murine oocytes treated prior to ovulation. Control animals were injected with intraperitoneal (ip) saline and the treatment animals were divided into three different dose schedules for ip cisplatin. After ovulation induction, the oocytes were processed for cytogenetic analysis and the chromosomes were counted. There were significantly less oocytes ovulated in the treatment groups when compared to control animals. There was a significant increase in hypohaploid cells counted in the 7.5 mg/kg treatment group, but the percentages of hyperhaploid and polyploid cells were not increased in the treatment groups. The cytogenetic effects of cisplatin in vivo and in vitro are reviewed.     

The effect of progesterone and RU486 on progesterone production in the ovulatory process of rats

To clarify the autoregulatory mechanism of progesterone (P) production in the ovulatory process, we examined the ovarian concentration of P 46 hrs after PMSG and the effects of P and RU486 (RU) injected 4-2 hrs before hCG administration on the serum concentrations of P and estradiol (E2), and ovarian 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activities in PMSG/hCG treated immature rats. The effect of RU on the number of ovulated ova was also studied. The ovarian concentration of P 46 hrs after PMSG was 0.96 +/- 0.03,ng/mg protein (mean +/- SEM). When P (100,mg/kg) was injected 2 hrs before hCG, ovarian 3 beta-HSD activities had significantly increased by 4 hrs after hCG. However, P at a dosage of 10 and 20,mg/kg had no effect on ovarian 3 beta-HSD activities. The administration of RU (20,mg/kg) 2 hrs before hCG significantly inhibited ovarian 3 beta-HSD activities measured 4 and 6 hrs after hCG (p less than 0.01 and p less than 0.05, respectively). In addition, the serum P concentration 4 hrs after hCG was significantly lower than that of the control (p less than 0.01). However, RU (20 mg/kg) in concomitant with hCG had no effect on ovarian 3 beta-HSD activities within 6 hrs after hCG. The suppression of ovarian 3 beta-HSD activities by RU was the concomitant reversed by the concomitant treatment with P (10 mg/kg). RU (10, 20, or 40 mg/kg) injected 2 hrs before hCG significantly reduced the number of ovulated ova (p less than 0.01, p less than 0.01 and p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Endocrine modifications associated with final oocyte maturation with gonadotropin-releasing hormone agonists vs. human chorionic gonadotropin in women undergoing intrauterine insemination

OBJECTIVE: To compare the hormonal profile when using triptorelin vs. human chorionic gonadotropin (hCG) to trigger ovulation in intrauterine insemination (IUI) cycles. STUDY DESIGN: Twenty-five patients who underwent 48 cycles were enrolled in a prospective, randomized, crossover study. After ovulation induction with recombinant follicle-stimulating hormone (FSH), couples were randomly allocated to a first cycle with a single dose of 0.2 mg of triptorelin or 5,000 IU of urinary hCG to trigger ovulation; if pregnancy did not occur, a second cycle was carried out, and the patient was crossed over to the other treatment group. Blood was collected the day of hCG/triptorelin administration and both days after IUI. RESULTS: Patients treated with triptorelin showed a significantly higher FSH and luteinizing hormone (LH) rise 24 hours after the injection when compared to hCG. Serum progesterone was significantly increased 48 hours after hCG administration, although estradiol levels were comparable between the groups. CONCLUSION: A higher LH and FSH peak than that induced by hCG was observed. Considering that serum progesterone levels were suboptimal in both protocols and taking into account the lower progesterone production in gonadotropin releasing hormone analogue cycles, corpus luteum supplementation in the luteal phase should be recommended for these patients.     

Luteinizing hormone bioactivity and variable responses to clomiphene citrate in chronic anovulation

In 18 women with infertility and chronic anovulation with normal gonadotropins, three different responses were observed to increasing doses (250 to 750 mg) of clomiphene citrate (CC). Follicle development and ovulation in 8, follicle development but no ovulation without human chorionic gonadotropin (hCG) in 6, and no response to CC in 4. Serum concentrations of bioactive luteinizing hormone (bioactive-LH), immunoactive (immunoactive-LH), follicle-stimulating hormone, and estradiol (E2) were measured and follicle growth was assessed by daily ultrasound. Findings were compared with 8 normal ovulatory controls. Folliculogenesis on CC therapy, based on our data, was 78%; however, only 44% ovulated spontaneously, 34% required hCG for follicle rupture. There were no apparent hormonal indicators to predict responders from nonresponders. The absence of an LH surge in the presence of follicles and sustained high E2 concentrations in 34% of patients may be associated with a decreased E2 sensitivity at the hypothalamic-pituitary level. Ultrasound easily identified patients who responded to CC with folliculogenesis but did not initiate an LH surge. Follicle rupture was achieved promptly by hCG administration.     

Effect of PGF2 alpha on ovulation and collagenolytic activities in the rabbit treated by indomethacin

To clarify a regulatory role of prostaglandins in collagenolytic enzyme activities of the follicular wall at ovulation, we studied whether the ovulation-blocking effect of indomethacin (IM) on hCG-stimulated rabbits could be reversed by an intravenous infusion of PGF2 alpha. The activities of BANA hydrolase and DNP peptidase were also measured by using synthetic substrates N-benzoyl-DL-arginine-2-naphthylamide HCl (BANA) and DNP-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg-OH (DNP peptide), respectively, every hour during the specific preovulatory period. The ovulation-blocking effect of IM (4mg/kg) was reversed by an intravenous infusion of PGF2 alpha (1.5 mg/kg) starting at 3 hr after the hCG injection. In the control rabbits treated with hCG(100 iu) alone, these enzymatic activities were increased toward ovulation with the peak level 7-9 hr after a hCG injection and decreased significantly at 10 hr. The preovulatory increase in these enzymatic activities disappeared following the ovulation-blocking doses of indomethacin. By the addition of the PGF2 alpha infusion, the IM-induced changes in BANA hydrolase and DNP peptidase activities were returned close to the control. It is suggested that the preovulatory increases in BANA hydrolase and DNP peptidase are mandatory for ovulation and PGF2 alpha facilitates ovulation through its regulatory action on these enzymes.     

补充黄体生成素对控制性超促排卵过程中卵巢慢反应患者临床结局的影响

目的:评价控制性超促排卵(COH)过程中卵巢慢反应患者补充重组黄体生成素(rLH)或人绝经期促性素(HMG)的有效性。方法:选取行长方案体外受精(IVF)或卵胞浆内单精子注射(ICSI)治疗的患者101例,均给予重组卵泡刺激素(rFSH)。将COH第7~8天血清黄体生成素(LH)持续正常者作为对照组(A组,n=34);将LH水平持续较低者随机分为两组,补充HMG组(B组,n=34)或rLH组(C组,n=33)。比较3组患者的临床结局。结果:B组患者的COH天数及rFSH用量均显著低于A组和C组(P0.05);C组的HCG日雌二醇(E2)水平、获卵数及双原核(2PN)受精率显著高于A组和B组(P0.05)。3组的促性腺激素(Gn)用量、HCG日LH、孕酮(P)、优质胚胎率及临床妊娠率比较,差异均无统计学意义(P0.05)。结论:对COH过程中卵巢慢反应的患者,补充HMG可减少COH天数及rFSH用量,降低治疗费用;补充rLH可改善卵巢的反应性,提高HCG日E2水平,增加获卵数及受精率,改善妊娠结局。     

Effects of momorcharins on ovarian response to gonadotropin-induced superovulation in mice

Alpha- and beta-momorcharins, which are abortifacient proteins isolated from Momordica charantia seeds, were tested for a possible effect on ovulation and plasma levels of ovarian steroids in mice induced to superovulate by PMSG and hCG. The plant proteins did not affect follicular recruitment and maturation as evidenced by ovarian histology and serum 17 beta-estradiol level. Both proteins diminished the number of oocytes ovulated when given on the day prior to or on the day of PMSG treatment, but not when given after the gonadotropin injection; they also increased the incidence of follicular atresia. The number of corpora lutea was slightly reduced by treatment with the proteins, but there was no long-term suppression of the serum progesterone level. After mating, animals which have previously been treated with the plant proteins underwent pregnancy resulting in a litter size similar to that of controls.     

Restoration of ovarian function by low nocturnal single daily doses of bromocriptine in patients with the galactorrhea-amenorrhea syndrome

Eighteen hyperprolactinemic patients with either amenorrhea or both galactorrhea and amenorrhea were treated with 2.5 mg of bromocriptine per day supplied at night. Ovulation occurred in 16 patients, and 8 wishing to conceive became pregnant. One of these patients became pregnant for a second time at a bromocriptine dose as low as 1.25 mg/day. Normal ovulatory cycles in four women receiving 2.5 mg of bromocriptine per day could also be maintained at 1.25 mg, despite the differences in the degree of prolactin (PRL) inhibition. Progesterone and luteinizing hormone levels measured for both bromocriptine levels were consistent with ovulatory cycles. The low, single dose of bromocriptine per day supplied at night decreased PRL levels, but normalization of blood levels did not always occur. Eighty percent of the patients ovulated with prolactin values over 25 ng/ml (normal value = less than 20 ng/ml).     

Progesterone protects oocytes from premature degeneration within the follicle

The present study was designed to determine the effects of gestrinone (R2323) in the process of follicle rupture and oocyte maturation and degeneration in an in vitro perfused rabbit ovary model. In the first experiment, R2323 at 10(2), 10(3), or 10(4) ng/ml was added to the perfusate of one ovary. The contralateral control ovary was perfused simultaneously with medium alone. Thirty minutes after the onset of perfusion, 50IU of human chorionic gonadotropin (hCG) was added to the perfusate of both ovaries. All ovaries exposed to R2323 plus hCG or hCG alone ovulated. The addition of R2323 to the perfusate did not affect the ovulatory efficiency of ovaries treated with hCG. No significant difference in the percentage of ovulated ova or follicular oocytes demonstrating germinal vesicle breakdown was seen with R2323 treatment. R2323 increased the degeneration rate of ovulated ova in a dose-dependent fashion. In the second experiment, in which experimental ovaries were perfused with R2323 (10(4) ng/ml) plus progesterone (10(3) ng/ml) and the control ovaries with R2323 (10(4) ng/ml) alone ovulation occurred in response to hCG. However, the addition of progesterone to the perfusate reduced the degeneration-inducing effect of R2323 on both ovulated ova and follicular oocytes. In conclusion, R2323 appears to act as an antiprogesterone, thereby promoting the degeneration of oocytes. The increased production of progesterone in the preovulatory follicle following the gonadotropin surge protects oocytes from premature degeneration within the follicles.     

Induction of ovulation with subcutaneous pulsatile gonadotropin-releasing hormone: correlation with body weight and other parameters

We treated 21 anovulatory infertile patients with subcutaneous pulsatile gonadotropin-releasing hormone (GnRH) administered via a syringe pump. Response to treatment was assessed by urinary estrogen excretion and ultrasound measurement of follicular growth. Ten patients ovulated and 8 subsequently conceived, for a total of 10 pregnancies. Human chorionic gonadotropin (hCG) was not administered routinely, but two patients required hCG to induce follicular rupture. The majority of the patients who conceived had a body mass index (BMI) of less than 21 and a luteinizing hormone (LH)/follicle-stimulating hormone ratio of less than 1. Conversely, those patients with either elevated BMI or LH or both generally failed to respond satisfactorily to this treatment. It is suggested that pulsatile GnRH is most likely to succeed in inducing ovulation if the BMI is less than 21 and the LH is normal, but is unlikely to be successful if there is both an elevated LH and a BMI of greater than 25. Between these two extremes, the response is variable and a therapeutic trial may be appropriate.     

LH improves early follicular recruitment in women over 38 years old

Although the capacity of recombinant FSH alone to induce folliculogenesis is undisputed, many believe that follicular recruitment in women over 38 years old could be improved by supplementing rFSH with human menopausal gonadotrophin (HMG). The present study sought to determine whether recombinant LH could reproduce the effect of HMG in women over 38 years during ovulation induction. Fifty-eight patients received rFSH (225 IU/day) supplemented with one ampoule of HMG (75 IU of FSH/75 IU of LH/HCG per day) for 5 days. Another 36 patients received rFSH (300 IU/day) supplemented with one ampoule of rLH (75 IU/day), also for 5 days. Both groups of patients received similar amounts of rFSH (1500 IU), LH/HCG (375 IU) and rLH (375 IU) and recruited a similar number of follicles as counted on day 6 (4.07 +/- 3.1 in the HMG group versus 3.7 +/- 3.2 in the LH group respectively) or on the day that human chorionic gonadotrophin (HCG) was indicated (6.5 +/- 2.7 versus 5.8 +/- 2.5 respectively). Ovarian stimulation was shorter, but not significantly so, in the group of patients receiving rFSH + HMG (10.5 +/- 1.7 days) than in the group of patients treated with rFSH +/- rLH (12 +/- 1.8 days). Significantly more MII oocytes were seen in the group treated with rFSH + rLH than in the group treated with rFSH + HMG (93.1 versus 75.3%, P < 0.05). With respect to pregnancy rates, 14/54 (26%) patients receiving rFSH + HMG and 16/34 (47%) patients receiving rFSH + rLH had a positive serum HCG. No significant difference in the number of miscarriages was observed between the two groups. In conclusion, the present results seem to indicate that rLH could be the HMG component that aids early follicular recruitment.     

Evaluation of two doses of recombinant luteinizing hormone supplementation in an unselected group of women undergoing follicular stimulation for in vitro fertilization

OBJECTIVE: To evaluate the efficacy of two doses of recombinant (r)LH, 75 IU (recommended) or 37.5 IU, for follicular stimulation and outcomes in a randomized cohort of IVF patients. DESIGN: Randomized, prospective analysis. SETTING: Private hospital incorporating an established IVF center. PATIENT(S): Women undergoing IVF who had a body mass index >18 or <35 and no abnormal karyotype, anovulation, oligomenorrhea, or any known endocrinopathy/illness. INTERVENTION(S): Pituitary desensitization was achieved with triptorelin (0.1 mg SC), and gonadotropin stimulation was performed with either rFSH alone (group A) or in combination with rLH in one of two doses: 37.5 IU (group B) or 75 IU (group C), daily. MAIN OUTCOME MEASURE(S): A range of endocrinologic, embryologic, clinical, and outcome parameters were evaluated. RESULT(S): With rLH supplementation there was a significant increase in the incidence of implantation (9% for rFSH only [group A] vs. 11% and 16% with 37.5 IU rLH and 75.0 IU rLH [groups B and C], respectively) and clinical pregnancy (19% vs. 23% and 31%) (P<.01 and P<.04, respectively), whereas there was no difference in the multiple pregnancy rates. There was a significant (P<.001) increase in the total units of rFSH used in proportion to the amount of rLH supplementation (2,645 U vs. 3,475 U and 3,681 U) and in the level of peripheral E(2) on the day of hCG administration (1,049 pg/mL vs. 1,640 pg/mL and 1,226 pg/mL) (P<.001). There was no significant between difference in mean age, numbers of oocytes recovered, basal and downregulation hormone levels, or the incidence of fertilization in the absence or presence of rLH supplementation, but a higher incidence of grade 1 to 2 embryos was observed when rLH was supplemented. CONCLUSION(S): After pituitary desensitization, there was an increase in the incidence of implantation, clinical pregnancy, and delivery rates in patients stimulated with rFSH supplemented with rLH.     

Effect of tetrahydrocannabinol on the hypothalamic-pituitary axis in the ovariectomized rhesus monkey.

Single doses of delta9-tetrahydrocannabinol (THC) (5.0, 2.5, 1.25, or 0.625 mg/kg) can decrease the levels of both luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the ovariectomized rhesus monkey. The inhibition of gonadotropins (50% to 88%) lasts for 6 to 24 hours depending upon the dose of THC. There are no great differences in the responses of the two gonadotropins to THC. The inhibition of gonadotropin levels by THC appears to be at the level of the hypothalamus, since both LH and FSH are released from the pituitary gland in response to LH- releasing factor in the presence of THC.     

Embryo loss, blastomere development and chromosome constitution after human chorionic gonadotropin-induced ovulation in mice and rats with regular cycles.

A dose of 7 IU human chorionic gonadotropin (hCG) given 14 h before the expected LH peak on proestrus significantly increased embryonic mortality in Swiss random-bred female mice to 55% of the number of corpora lutea. The use of luteinizing hormone-releasing hormone in a similar injection protocol did not induce embryonic death. The effect found in Swiss random-bred mice resembles that of a dose of 20 IU hCG in the rat. Afternoon-day-4 mouse embryos contained 39.1 +/- 12.6 nuclei after hCG-induced ovulation compared to 46.2 +/- 16.6 nuclei after spontaneous ovulation. For early-day-5 embryos of the rat, these figures were 34.2 +/- 10.1 and 31.7 +/- 8.4, respectively (mating was early on day 1). Numerical chromosome errors were estimated in secondary oocytes of the mouse and early-day-5 embryos of the rat. Compared with data from the literature, hCG seems to induce some extra meiotic nondisjunction in the rat only. Combining all genetic and physiological data, the loss of fecundity after hCG-induced ovulation is a maternal effect.     

The relationship between prostaglandins and histamine in the ovulatory process as determined with the in vitro perfused rabbit ovary

The process of follicle rupture has been described as an inflammatory reaction in which prostaglandins (PGs) and/or histamine may be involved. With an in vitro perfused rabbit ovary preparation, experiments were carried out for determination of whether a relationship exists among PGs, histamine, and ovulation. PGF2 alpha alone was capable of inducing ovulation when added to the perfusion fluid at 1, 10, and 100 ng/ ml. Effectiveness in achieving ovulation varied directly with the dosage; however, the ovulatory efficiency of PGF2 alpha-treated ovaries was lower than that of ovaries exposed to human chorionic gonadotropin (hCG, 100 IU). PGF2 alpha-induced ovulation could not be blocked by the H2 receptor antagonist, cimetidine. The PG synthesis inhibitor, indomethacin, did not prevent histamine-induced ovulation. Ovulation induced by hCG was partially blocked by the administration of indomethacin; however, the concomitant administration of cimetidine was not associated with further reduction in ovulation. In all but one experimental group, the majority of ovulated ova did not progress beyond the intact germinal vesicle stage unless the ovaries had been exposed to hCG. On the basis of these experiments, PGs and histamine do not appear to be interdependent in their effects on the ovulatory process in vitro.     

Effect of clomiphene citrate on in vitro ovulated ova

The effect of clomiphene citrate (CC) on developmental capacity of ovulated ova was studied with isolated in vitro perfused rabbit ovaries. Fifty-two ovulated ova were recovered from ovaries perfused with human chorionic gonadotropin (hCG) in a medium containing CC and 45 ova from ovaries perfused with hCG in a CC-free medium. Ova were cultured and inseminated with capacitated sperm and observed serially for evidence of fertilization and stage of development. CC did not affect the fertilization of ovulated ova. However, the percentage of ova which had reached the morula stage by 60 hours was significantly reduced in the CC-treated (15.4%) group, compared with the control group of ovaries (48.9%). A significant percentage of inseminated ova from CC-treated ovaries (65.4%) showed evidence of degeneration, as compared with control ovaries (37.8%). Thus, a partial loss of developmental capacity may explain the discrepancy observed between the rate of successful ovulation induction and the establishment of pregnancy associated with CC administration in humans.     

Daily administration of the progesterone antagonist RU 486 prevents implantation in the cycling guinea pig.

Since progesterone is required to prepare the endometrium for implantation of an embryo, a progesterone antagonist may inhibit nidation and thus prevent pregnancy. We addressed this possibility in the guinea pig, the small laboratory animal whose reproductive physiology most resembles that of women. Daily administration of the antiprogestin RU 486 (0, 1, 2, or 3 mg/kg, subcutaneously) for 9 days after mating inhibited implantation in a dose-dependent fashion. When this compound was given daily throughout the estrous cycle, cyclic vaginal changes, ovulation, and mating were suppressed in up to 17%, 28%, and 55% of animals, respectively. Two of seven mated female animals receiving RU 486, 1 mg/kg/day, had implantation sites. Nidation was completely blocked at higher doses. Thus daily antiprogestin administration prevented pregnancy in sexually active, normally cycling guinea pigs. A similar strategy using a daily antinidatory dose of an antiprogestin may offer a novel approach to human fertility control.     

Ibuprofen modulation of human chorionic gonadotropin-induced ornithine decarboxylase activity and ovulation in the rabbit ovary

The activity of ornithine decarboxylase, the rate-limiting enzyme in polyamine synthesis, increases as granulosa cells proliferate in developing follicles. Both luteinizing hormone and prostaglandins stimulate ornithine decarboxylase activity. Here, we sought to determine the relative contributions of both trophic stimuli to ornithine decarboxylase activity in the preovulatory rabbit ovary. Baseline ovarian ornithine decarboxylase activity, determined by measuring the release of CO2 from (1-14C)-ornithine, was 13.4 +/- 1.27 (mean +/- SD) pmol of carbon dioxide per hour per milligram of protein. Treatment with ibuprofen, a prostaglandin synthetase inhibitor, led to a significant (p less than 0.05) decrease in the baseline ovarian ornithine decarboxylase activity (4.7 +/- 0.29 pmol of carbon dioxide per hour per milligram of protein). Administration of human chorionic gonadotropin (hCG) (50 IU/kg intramuscularly) to adult rabbits (2.5 to 3.5 kg) elicited a 1,200% elevation of ovarian ornithine decarboxylase activity 5 hours after injection; there was a return to baseline by 11 hours after injection. Stimulation with human chorionic gonadotropin led to ovulation in 22.2%, 25%, and 60% of rabbits at 7, 9, and 11 hours after treatment, respectively. Single-dose ibuprofen treatment (5 mg/kg intramuscularly), 7 hours after human chorionic gonadotropin administration inhibited ovulation and elevated ovarian ornithine decarboxylase activity. These results indicate that ibuprofen effectively inhibits ovulation in hCG-stimulated rabbit ovaries in the presence of a significant (p less than 0.001) elevation in ovarian ornithine decarboxylase activity. Thus, different intracellular mechanisms are involved in the prostaglandin modulation of basal and hCG-stimulated cells during the course of preovulatory follicle maturation.