I.F. Lau

Pregnancy blockade by indomethacin, an inhibitor of prostaglandin synthesis: Its reversal by prostaglandins and progesterone in mice

Abstract Results in the present study clearly indicate that subcutaneous injections of indomethacin (150 μg/day) to the pregnant mice on days 1,2,3 and 4 of pregnancy terminate pregnancy in 100 percent of the animals. A single injection of 225 μg/day/mouse on day 2 of pregnancy results in 100 percent inhibition of pregnancy. Since this dose of indomethacin was not as effective when given either on day 1, 3 or 4 of pregnancy, it is likely that indomethacin acts through a tubal disturbances by suppressing prostaglandins for the retention of the ova. Progesterone (0.75 mg/day) and prostaglandins E 2 and F 2α (50 μg/injection) reverse the anti-implantation effect of indomethacin. Other possibilities are that indomethacin exerts its antifertility effect i) by delaying nidation, ii) by an indirect or direct effect on the developing embryos, iii) by causing a hormonal imbalance which results in either progesterone deficiency or by suppressing the circulating level of luteinizing hormone.

Variations in serum androgens, estrogens, progestins, gonadotropins and prolactin levels in male rats from prepubertal to advanced age

The concentrations of blood serum steroids from 12 to 450 days old male rats were determined by radioimmunoassay. Testosterone (T) was low (270 pg to less than 1 ng/ml) until day 42; adult levels (3--4 ng/ml) were attained by day 62 and declined tradually with advanced age. 5 alpha-dihydrotestosterone (5 alpha-DHT) did not change markedly (90--160 pg/ml) from prepubertal to advanced age. Except for a small peak on day 22, androstenedione (delta 4 A) levels ranged between 400-500 pg/ml in the adult but declined in older males. Progesterone (delta 4 P) rose steadily to a mean of 5.46 ng/ml at 52 days of age and dropped thereafter. High levels of estrone (268 +/- 38 pg/ml) and estradiol-17 beta (2.76 +/- 0.28 ng/ml) in 12 days old males are in contrast to the low estrogens (20-35 pg/ml) in adult animals. Both T/5 alpha-DHT and total T/estrogen ratios were low before puberty, increased in adults and decreased towards old age. The interplay between gonadotropin and prolactin, which exhibited reciprocal changes in the regulation of steroid production by the gonads with age, are discussed.

The inhibition of the conversion of testosterone into 5α-dihydrotestosterone in the reproductive organs of the male rat

The inhibition of testosterone 5alpha-reductase activity by 3-oxo-4-androstene-17beta-carboxylic acid in the male reproductive organs of the rat was demonstrated in vitro. The medium for incubation of caput epididymis showed the highest concentration of 5alpha-dihydrotestosterone (5alpha-DHT) whereas the highest concentration of testosterone (T) was recorded in medium for incubation of decapsulated testis after two hours of incubation. The 3-oxo-4-androstene-17beta-carboxylic acid (1.58 X 10(-5)M) inhibited the conversion of T to 5alpha-DHT in all the organs tested (testis, caput and cauda epididymis and ventral prostate) under identical incubation conditions.

Effect of exogenous estradiol and progesterone on the uterine tissue levels of prostaglandin F2α(PGF2α) in ovariectomized mice

Abstract Mice ovariectomized for 14 days were treated for 6 days with estradiol and/or progesterone. Both the steroids were effective in increasing the levels of PGF 2α in the uterine tissue, but the treatment with progesterone for 3 days followed by 3 days of estrogen resulted in a highly significant production of PGF 2α . It is concluded that for the production of PGF 2α both estrogen and progesterone are necessary and that the pretreatment with progesterone followed by estrogen results in the maximum production of PGF 2α .

Prostaglandin mediated action of IUD's (II) F-prostaglandins (PGF) in the uterine horn of pregnant rats and hamsters with intrauterine devices

Abstract The present investigation clearly shows that the presence of an IUD is associated with uterine hypertrophy and an increased production of F-prostaglandins. In the hamster, a bilateral IUD produced a significant increase in peripheral plasma PGF, presumably of uterine origin and an increased uterine PGF content at 2230 – 2300 h of day 4 of pregnancy. Earlier sampling (1530 – 1600 h) on the same day of pregnancy did not reveal any significant difference. In the rat, uterine PGF content and concentration, as well as PGF levels in uterine venous plasma were all increased in the IUD bearing horn from day 4 pregnant animals at 1530 – 1630 h in comparison to the contralateral control horn.

Age dependent changes in the sperm population and fertility in the male rat

The entire reproductive tract was devoid of spermatozoa during the first 42 days of life. The first appearance of spermatozoa was detected in the caput epididymidis of 45-day old rats and in the cauda epididymidis of 52-day or older rats. The number of spermatozoa in the reproductive tract increased with age. The sperm population reached its maximum in the caput epididymidis by day-72 and in the caud epididymidis by day-100. These high levels were maintained beyond 450 days of age. Fifty-two day old male rats failed to impregnate the female. Although male rats of 62-500 days were able to sire litters, the peak reproductive period was found to be between days 100-270, during which the number of young per litter ranged between 11-16. Towards advanced age the litter size dropped to 8. The lower fertility in the males of advanced age might be due to a lower proportion of fertile spermatozoa, a change in the physiology of epididymis or an altered sperm transport in the reproductive tract.

Temporal changes in circulating steroids during prostaglandin F2α induced abortion in the rat and rabbit

The serum levels of progesterone (delta4P), 20alpha-dihydroxyprogesterone (20alpha-DHP), pregnenolone (delta5P) and estrogens at different time intervals during Silastic-PVP-PGF2alpha tube induced midterm abortion were measured by radioimmunoassay in the rat and rabbit. The concentrations of delta4P and delta5P were significantly reduced but that of 20alpha-DHP was increased significantly as early as 20 hours after PGF2alpha treatment. Estrogen levels showed slight reduction in the rat but remained unchanged in the rabbit. The ratio of delta4P/20alpha-DHP recorded were 2.15 and 16.86 (pretreatment) and dropped to 0.08 and 0.37 (60 hours after insertion of Silastic-PVP-PGF2alpha tube) in the rat and rabbit, respectively. These findings suggest that the suppression of circulating delta4P by PGF2alpha might be the result of a reduced precursor (delta5P) pool size as well as changes in the enzymic systems responsible for the formation and conversion of delta4P. It is still to be determined if changes in estrogen levels in the rat have any bearing on abortion induced by PGF2alpha.

Determination of Seven Unconjugated Steroids in the Blood and Seminal Plasma of the Fertile Male Rabbit

Seven unconjugated steroids were measured in the blood and seminal plasmas of fertile male rabbits by radioimmunoassay. The blood plasma testosterone concentration was 4--5 times that of the seminal plasma. Dehydroepiandrosterone, estrone and 17beta-estradiol were found in measurable amounts in the blood plasma; however, these steroid levels were slightly lower in seminal plasma. Androstenedione and 5alpha-dihydrotestosterone were present in equal quantities in both the seminal and blood plasmas. By contrast, seminal plasma pregnenolone level was about twice that of the blood plasma. The determination of seminal plasma steroids may lend itself as a complementary assessment to blood steroid determinations for the evaluation of the normal function of various reproductive organs.

Termination of pregnancy in rabbit and mouse by Trichosanthin.

The effects of a single intraperitoneal injection of Trichosanthin in 6-, 10-, 17- or 22-day pregnant rabbit (2 mg/rabbit) or in 11-day pregnant mouse (50 micrograms/mouse) were studied. Trichosanthin induced abortion in 100% of the 17- or 22-day pregnant rabbits within 48-72 hours and decreased circulating progesterone (delta 4P)concentrations with 24 hrs. On the other hand, the same dose failed to terminate pregnancy in 6- or 10-day pregnant rabbits and caused no significant changes in circulating delta 4P levels. Exogenous delta 4P or prolactin + human chorionic gonadotropin given twice daily failed to reverse the Trichosanthin-induced termination of pregnancy. However, this resulted in a delay of fetal expulsion. It is concluded that Trichosanthin-induced termination of pregnancy is not solely a result of luteolysis but is likely to be due to its toxic effects on placenta, embryo or both. A dose of 50 microgram Trichosanthin given to 11-day pregnant mice resulted in a termination of pregnancy within 96 hours and also a significant decrease in delta 4P levels in 24 hrs. The ratio of delta 4P to 20 alpha-dihydroprogesterone was also decreased steadily after Trichosanthin administration.

FURTHER STUDIES ON THE TRICHOSANTHIN-INDUCED TERMINATION OF PREGNANCY

Trichosanthin (TCS), a protein from the root extract of Trichosanthis kirilowii, terminated pregnancy when injected once in 15-day pregnant rabbits (2 mg/doe) but failed to interrupt pregnancy in 12-day pregnant rabbits even at higher doses. In vitro release of progesterone (delta 4P) from the maternal or fetal placental tissue into the incubation medium was not affected by TCS. When the distribution of 125I-TCS was traced in 12-day pregnant mice, persistently higher concentration of 125I-radioactivity was detected in the kidney. By contrast, in other organs, including the reproductive organs, blood serum and amniotic fluid 125I-radioactivity declined between 12-48 h after treatment. The low 125I-radioactivity in both the reproductive organs and amniotic fluid suggests a possible barrier between the embryo and maternal blood. It is suggested that TCS might be acting directly on the placental unit, causing fetal death and dislodging of the placenta. Administration of TCS to PD-19 mice or PD-28 rabbits resulted in premature delivery. This effect of TCS in pregnant mice was comparable to the administration of PGF2 alpha.