Effect of aqueous leaf extract of Dalbergia sissoo Roxb. on spermatogenesis and fertility in male mice.

OBJECTIVES: Antifertility effects of Dalbergia sissoo in male mice were investigated. METHODS: Adult Parkes strain male mice were orally administered aqueous leaf extract of Dalbergia sissoo (50 and 100 mg/kg body weight/day) or distilled water or no treatment (controls) for 35 days (n = 5/group). Motility, viability and number of spermatozoa in the cauda epididymidis; testis histology; serum level of testosterone; and toxicological parameters were evaluated. To assess reversibility, more mice were treated with 100 mg/kg body weight of Dalbergia sissoo or distilled water (n = 5/group) for 35 days and sacrificed 56 days later. Fertility was also assessed separately. RESULTS: Histologically, testes of Dalbergia-treated mice showed dissimilar degenerative changes in the seminiferous tubules. Significant reductions were noted (i) in epididymal sperm motility, viability and number, and (ii) in serum level of testosterone in Dalbergia-treated mice compared to controls. However, serum levels of alanine aminotransferase, aspartate aminotransferase and creatinine, and haematological parameters were not affected. Also libido of Dalbergia-treated males showed no change, but their fertility was markedly suppressed. By 56 days of treatment withdrawal, alterations induced in the above parameters returned to control levels. CONCLUSIONS: Dalbergia sissoo treatment caused reversible suppression of spermatogenesis and fertility in P mice, without eliciting detectable toxic effects.

Reversible antispermatogenic and antisteroidogenic activities of Feronia limonia fruit pulp in adult male rats.

OBJECTIVE: To explore the antispermatogenic and testicular antisteroidogenic activities of Feronia limonia fruit pulp southern India. METHODS: Fourty Wistar male albino rats (Rattus norvegicus) were equally divided into four groups. Experimental groups were administered with the ethanolic extract of Feronia limonia (F. limoni) fruit pulp at doses of 250 and 500 mg/kg body weight once daily for 55 days. All treated rats had corresponding recovery groups. At the end of each treatment periods, various spermatological indices, tissue biochemicals and testicular enzymes levels were analysed. Blood profiles were also estimated. RESULTS: Compared with the control, the F. limonia fruit pulp at both dose levels did not decrease body weight, which were associated with decline in epididymal sperm count, motility, viability and increased percent of abnormal sperm. Further, F. limonia fruit pulp at 500 mg/kg body weight markedly reduced the epididymal and testicular protein content by 24.58% and 29.86%, respectively, as well as the glucose-6-phosphate dehydrogenase and Δ(5)-3ß-hydroxy steroid dehydrogenase) levels by 42.82% and 38.08%, respectively, while a significant elevation was observed in testicular cholesterol and ascorbic acid content. A gradual recovery of all parameters was observed after 55 days of treatment withdrawal. No significant alterations in haematological indices were observed. CONCLUSIONS: The present findings indicate that F. limonia fruit pulp may have reversible antispermatogenic and antisteroidogenic properties, and could partially support the traditional use as male contraceptive.

Toxicological studies of momordica charantia linn seed extracts in male mice / Estudios toxicológicos de los extractos de la semilla de Momordica charantia Linn en ratones macho

An attempt to find out the male contraceptive molecule of plant origin, the extracts of seeds of Momordica charantia were tested in male mice. Petroleum ether, chloroform and ethanolic extracts of Momordica charantia were administered at the dose level of 25mg/100gm body weight to the albino mice for 48 days intraperitoneally. All the extracts showed antispermatogenic effect as the number of spermatogonia, spermatocytes, spermatids and spermatozoa were decreased. The increase in the weight of epididymis, prostate gland, seminal vesicle and vas deferens indicates clearly the androgenic property of these extracts. After subjecting to preliminary phytochemical screening ethanol extract showed positive tests for alkaloids, flavonoids, glycosides, phenols, tannins, oils and fats. Out of the three extracts tested, the ethanol extract seems to be more potent in its contraceptive and androgenic activities.

En un intento por descubrir la molécula de anticoncepción masculina de origen vegetal, fueron probados los extractos de semillas de Momordica charantia en ratones machos. Extractos de éter de petróleo, cloroformo y etanol de Momordica charantia fueron administrados en dosis de 25mg/100g de peso corporal a ratones albinos de 48 días por vía intraperitoneal. Todos los extractos mostraron un efecto antiespermatogénicos, con reducción del número de espermatogonias, espermatocitos, espermátidas y espermatozoides. El aumento de peso del epidídimo, próstata, vesículas seminales y conductos deferentes indica claramente la propiedad androgénica de estos extractos. Después de someter el extracto de etanol a la detección preliminar fitoquímica se observaron resultados positivos para alcaloides, flavonoides, glucósidos, fenoles, taninos, aceites y grasas. De los tres extractos probados, el extracto de etanol parece ser más potente en sus actividades de anticonceptivas y androgénicas.

[Anti-fertility effect of Tongbi composition and its reversibility in male rats].

OBJECTIVE: To study the anti-fertility effect of maximum-dose Tongbi Composition and its reversibility in male rats. METHODS: Thirty-six male SD rats were equally randomized into a control group and a medication group, the former given normal saline at 10 ml/(kg x d), while the latter treated with Tongbi Composition at 10 g/(kg x d), both for 60 days. Half the rats of each group were sacrificed randomly at the cessation of treatment, and the rest killed at 72 days after it. The relative testis weight, testis volume, sperm concentration and sperm motility were measured, and the pathological changes in the testicular tissue observed under the optical microscope. RESULTS: After 60 days of treatment, no statistically significant differences were found between the two groups in the relative testis weight, testis volume and sperm concentration (P > 0.05) , and the sperm motility of the medication group dropped to zero, but it was restored to normal at 72 days after drug withdrawal. Almost no lesions were observed in the testis tissue of the medication group. CONCLUSION: The short-term use of Tongbi Composition at the maximum clinical dose has an obvious anti-fertility effect, but it is reversible.

Inhibition of hyaluronidase activity of human and rat spermatozoa in vitro and antispermatogenic activity in rats in vivo by Terminalia chebula, a flavonoid rich plant.

Our interest in development of hyaluronidase inhibitors as male antifertility agents led to identification of Terminalia chebula (T. chebula) plant with hyaluronidase (HAase) inhibitory activity of human spermatozoa ( approximately 93% inhibition) and rat caudal epididymal spermatozoa ( approximately 86% inhibition) in vitro at 30 mg/ml. We further demonstrated inhibition of hyaluronidase activity of testis and epididymal spermatozoa in vivo coincident with antispermatogenic activity and contraceptive efficacy of TC extract administered at 50 and 100mg/kg/day orally for 60 days in male albino rats. The significant decrease in motility, count and increase in morphological abnormalities of epididymal spermatozoa and severe reduction in fertility (-100%) of male rats treated with T. chebula fruit extract at 100mg/kg dose could be attributed to either direct effect on testis or direct or indirect interference with sperm maturation in epididymis, and/or inhibition of testicular and epididymal sperm hyaluronidase enzyme in vivo probably caused by flavonoids like tannins present in T. chebula.

Effect of methanolic extract of Dendrophthoe falcata stem on reproductive function of male albino rats.

In spite of the considerable development in contraceptive technology, search for male antifertility agents in plants continues to be a potential area of investigation. Many plants have been known to possess antifertility activity, but limited attempts have been made to scientifically evaluate these claims. Hence the purpose of this study was to evaluate the antifertility and reproductive toxicity potential of Dendrophthoe falcata (Loranthaceae) in male Wistar rats. An oral 70% methanolic extract of stem of D. falcata at a dose level of 100 mg/kg wt/day fed to male albino rats for 60 days did not decrease body weight, while the testes and epididymides were significantly reduced, and the seminal vesicles and ventral prostate also showed a significant reduction (P < 0.01). Treated animals showed a notable depression of spermatogenesis. As a result of 100 mg/kg extract feeding, the preleptotene spermatocytes, secondary spermatocytes, step-19 spermatids and the mature Leydig cells decreased by 74.36%, 80.03%, 79.87%, 32.37%, respectively. At this dose Leydig cell nuclear area and cytoplasmic area, as well as the cross sectional surface area of Sertoli cells, were significantly reduced (P < 0.001) when compared to controls. The reduced sperm count and motility resulted in 100% negative fertility at 100 mg/kg dose level. A significant fall in the total protein and sialic acid content in the testes, epididymides, seminal vesicle and ventral prostate, as well as in the glycogen content of testes was also observed. The level of serum protein, cholesterol, billirubin, SGOT, SGPT, blood urea, and hematological indices were unaltered. In conclusion, Dendrophthoe falcata brought about the inhibition of spermatogenesis.

Antispermatogenic, antiandrogenic activities of Albizia lebbeck (L.) Benth bark extract in male albino rats.

Methanolic extract of Albizia lebbeck bark when administered orally at the dose level of 100 mg/rat/day to male rats of proven fertility for 60 days did not cause any significant loss in their body weights but the weights of reproductive organs, i.e. testis, epididymides, seminal vesicle and ventral prostate were decreased in a significant manner when compared to controls. Sperm motility as well as sperm density were reduced significantly which resulted in reduction of male fertility by 100%. Marked decline in the germ cell population was noticed. Population of preleptotene, pachytene, secondary spermatocytes and step-19 spermatid were declined by 60.86%, 65.81%, 71.56% and 66.55%, respectively. Cross-sectional surface area of sertoli cells as well as the cells counts were found to be depleted significantly. Leydig cells nuclear area and number of mature Leydig cells were decreased by 60.03% and 51.56%, respectively. Serum testosterone levels showed significant reduction after A. lebbeck extract feeding. Oral administration of the extract did not affect red blood cell (RBC) and white blood cell (WBC) count, haemoglobin, haematocrit and glucose in the blood and cholesterol, protein, triglyceride and phospholipid in the serum. In conclusion, A. lebbeck bark extract administration arrests spermatogenesis in male rats without noticeable side effects.

Antispermatogenic activity of Morinda lucida extract in male rats.

AIM: To investigate the effect of Morinda lucida Benth (Rubiaceae) on the reproductive activity of male albino rats. METHODS: Two groups of rats were treated with 400 mg/(kg .d) of Morinda lucida leaf extract for 4 and 13 weeks, respectively. The control rats received the vehicle. All the treated rats had corresponding recovery groups. At the end of each experimental period, animals were killed and organ weights, sperm characteristics, serum testosterone levels, histology of the testes and fertility were assessed. RESULTS: Morinda lucida leaf extract did not cause any changes in body and somatic organ weights, but significantly increased the testis weight (P 0.05). The sperm motility and viability, and the epididymal sperm counts of rats treated for 13 weeks were significantly reduced (P 0.05). Sperm morphological abnormalities and serum testosterone levels were significantly increased (P 0.05). There were various degrees of damage to the seminiferous tubules. The extract reduced the fertility of the treated rats by reducing the litter size. Reversal of these changes, however, occurred after a period of time. CONCLUSION: The extract of Morinda lucida has reversible antispermatogenic properties.

[Progress in research on triptolide].

To further understand triptolide, this paper has introduced the pharmacology, pharmacokinetics, toxicity, the clinic application and semi-synthesis of triptolide on basis of importance and significant contents of reference which have been consulted in the past twenty years. Presently triptolide and Tripterygium wilfordii have been a hot spot of modernization of Chinese traditional medicine. It is very important to develop a new dosage form of high effect and low toxicity by making use of advanced technology according to its characteristics.

Antispermatogenic and hormonal effects of Crotalaria juncea Linn. seed extracts in male mice.

AIM: To evaluate the antifertility activity of various extracts of Crotalaria juncea seeds in male mice. METHODS: Adult male mice were gavaged the petroleum ether, benzene and ethanol extracts of C. juncea seeds, 25 mg x (100g)(-1) x day(-1) for 30 days. On day 31 the animals were sacrificed by cervical dislocation and the testes, epididymis, vas deferens, seminal vesicles, prostate gland, bulbourethral gland and levator ani were dissected out and weighed. The organs were processed for biochemical and histological examination. RESULTS: In petroleum ether, benzene and ethanol extracts treated rats, there was a decrease in the weights of testis and accessory reproductive organs. The diameters of the testis and seminiferous tubules were decreased. Spermatogonia, spermatocytes and spermatids in the testis and the sperm count in cauda epididymis were also decreased. There was a significant reduction in the protein and glycogen contents and an increase in the cholesterol content in the testis, epididymis and vas deferens. Of the 3 extracts, the ethanol extract appeared to be the most potent in antispermatogenic activity. When the ethanol extract was tested in immature male mice, there was an antiandrogenic effect as the weights of accessory organs were reduced. CONCLUSION: The various extracts of C. juncea seeds arrest spermatogenesis and are likely to have an antiandrogenic activity.

A comprehensive evaluation of the reproductive toxicity of Quassia amara in male rats.

Chloroform extracts of the bark of Quassia amara in different dilutions was used to assess its impact on the male reproductive system of albino rats. Single daily intramuscular injections of the extract for 15 days resulted in a significant reduction in the weight of testis and epididymis but not that of the seminal vesicles and prostate (all lobes). A marked decrease in the sperm count, motility, viability was also observed in sperm collected from the cauda epididymis of treated animals. A number of abnormalities like double heads, double tails, detached heads and fragile tails were frequently seen. Epididymal alpha-glucosidase activity was drastically reduced. However, prostatic acid phosphatase activity and citric acid levels and seminal vesicle fructose concentrations remained unchanged following treatment. Thus, it appears that the prime site of action is at the level of both the testis and the epididymis. Examination of the blood showed that cell counts and hemoglobin levels were in the normal range. Bilirubin, SGPT, SGOT, protein and urea were also not altered by the herbal extract. From the selective action on the male reproductive tract we suggest that the chloroform extract of the bark of Quassia amara has potential for use as an antifertility agent.

Inhibition of Ca(2+) channels in mouse spermatogenic cells by male antifertility compounds from Tripterygium wilfordii Hook. f.

The male antifertility effect of a water-chloroform extract (GTW) from the root xylem of Tripterygium wilfordii has attracted worldwide interest. In the present study, by using whole-cell recording, the effects of GTW and two isolated monomers from GTW, demethylzeylasteral and L-epicatechin, on the T-type Ca(2+) channels in mouse spermatogenic cells were investigated. The results showed that each of them concentration-dependently and partially reversibly inhibited T-type Ca(2+) current in the cells. The IC(50) of GTW and demethylzeylasteral were approximate, while L-epicatechin inhibited the channels at a much higher concentration. The voltage dependence of the inhibitory effect and the changes in activation and inactivation time constants after application of these compounds were also examined. These data suggest that the inhibition of T-type Ca(2+) currents could be responsible for the antifertility activity of these compounds.

Antifertility studies of Colebrookia oppositifolia leaf extract in male rats with special reference to testicular cell population dynamics.

Oral feeding of male rats with the ethanolic leaf extract of Colebrookia oppositifolia at dose levels of 100 and 200 mg/kg for 8-10 weeks did not cause body weight loss, while the weights of testes and epididymides were significantly decreased. Seminal vesicles and ventral prostate showed a significant reduction at the higher dose only. Treated animals showed a notable depression of spermatogenesis. Following 100 and 200 mg/kg extract feeding, the preleptotene spermatocytes were decreased by 46.5 and 39.8%, the secondary spermatocytes by 13.4 and 12.7%, the step-19 spermatids by 36.6 and 35.2%, and the mature Leydig cells by 31.2 and 39.5%, respectively. At both dose levels, the seminiferous tubule diameter, Leydig cells nuclear area and cytoplasmic area, as well as the cross-sectional surface area of Sertoli cells, were significantly reduced (P<0.001) when compared to controls. Reduced sperm count and motility resulted in 100% negative fertility at 200 mg/kg dose level. A significant fall in the total protein and sialic acid content and acid phosphatase enzyme activity of the testes, epididymides, seminal vesicle and ventral prostate, as well as in the glycogen content of testes, was also observed at both dose levels in comparison with controls.

Reversible antispermatogenic and antifertility activities of Mondia whitei L. in male albino rat.

Chronic administration of Mondia whitei L. root bark extract (400 mg/kg/day) for 55 days caused testicular lesions resulting in the cessation of spermatogenesis, degenerative changes in the seminiferous tubules and epididymides. The wet weight of the seminal vesicle increased, whereas the weights of testes, epididymides and ventral prostate were unchanged. The treatment also resulted in a partial antifertility effect, and an increase in the protein content of the testes and epididymides. The cholesterol contents of the testes were significantly elevated after 55 days, whereas testosterone and 17beta-oestradiol contents of the testes were unchanged. Serum protein was elevated but serum testosterone was unchanged. A recovery period resulted in normal spermatogenesis and fertility, suggesting reversible antispermatogenic and antifertility effects of the plant.

Long-term effects of triptolide on spermatogenesis, epididymal sperm function, and fertility in male rats.

Prior studies had suggested that triptolide, a diterpene triepoxide isolated from a Chinese medicinal plant, might be an attractive candidate as a post-testicular male contraceptive agent. Despite the promise that triptolide would not affect testis function, nagging concerns remained that a delayed onset of testicular effect might exist. The objectives of this study were to assess the effects of relatively longer treatment duration of triptolide on fertility, spermatogenesis, and epididymal sperm pathophysiology; and to evaluate the reversibility of these effects after the cessation of treatment. Adult male Sprague-Dawley rats were fed daily with either 30% gum acacia as a vehicle control (n = 12) or 100 microg/kg body weight (BW) of triptolide for 82 days (n = 12) followed by a recovery period of up to 14 weeks (n = 6). At the end of the treatment period, all rats treated with triptolide were sterile. Cauda epididymal sperm content decreased by 84.8% and sperm motility was reduced to zero. In addition, virtually all cauda epididymal sperm in the triptolide-treated group exhibited severe structural abnormalities. The most striking changes observed were head-tail separation, premature chromatin decondensation of sperm nuclei, a complete absence of the plasma membrane of the entire middle and principle pieces, disorganization of the mitochondrial sheath, and aggregation of many sperm tails. Longer treatment duration of triptolide also affected spermatogenesis, with marked variability in the response of individual animals. The degree of damage ranged from apparently normal-looking seminiferous tubules to flattened seminiferous epithelium lined by a single layer of cells consisting of Sertoli cells and a few spermatogonia. Affected tubules exhibited intraepithelial vacuoles of varying sizes, multinucleated giant cells, germ cell exfoliation, and tubular atrophy. Recovery occurred as early as 6 weeks after cessation of treatment. By 14 weeks, 4 out of 6 triptolide-treated males were fertile and the females that were impregnated by 3 out of 4 triptolide-treated male rats produced apparently normal litters. These results suggest that triptolide has 2 phenotypic effects on mature and maturing germ cells. The first action appears earlier and manifests mainly in epididymal sperm. The second action presumably is directly on germ cells in testis and causes a variable impairment of spermatogenesis that may not be completely reversible. It is unclear if the earlier effect is a delayed manifestation of subtle testicular injury or post-testicular action.

Aspects of the male reproductive toxicity/male antifertility property of andrographolide in albino rats: effect on the testis and the cauda epididymidal spermatozoa.

Previous work has shown that Andrographis paniculata leaf, when fed to male albino rats, causes the arrest of spermatogenesis. The present study was undertaken to investigate whether andrographolide, one of the major constituents of this plant, is responsible for such an effect. The compound was administered to 3-month-old male Wistar albino rats at two dose levels, for 48 days. Fertility tests, analysis of the counts, motility and abnormalities of the cauda epididymidal spermatozoa, and histopathological-evaluation of the testis were carried out. The results showed that sperm counts decreased, the spermatozoa were not motile, and several of them possessed abnormalities. The seminiferous epithelium was-thoroughly disrupted and in the seminiferous tubules, fully differentiated spermatozoa were far too limited; cells in the divisional stages were prevalent; multinucleate giant cells were abundant and Leydig cells appeared intact. It is inferred that andrographolide could affect spermatogenesis by preventing cytokinesis of the dividing spermatogenic cell lines. The multinucleate giant cells are comparable to the symplasts generated by cytochalasin-D and ursolic acid due to action at stages V-VII of the spermatogenic cycle. Sertoli cell damage and spermatotoxic effects are also apparent. Thus, the study points to a male reproductive toxic effect of this compound when used as a therapeutic; the study also confirms the possible prospective use of andrographolide in male contraception.

[Comparative study on the effect of gossypol and T7 on human spermatozoa ATPase activity].

OBJECTIVE: A comparative study was carried out on the effect of two kinds of male contraceptive drugs, gossypol and the abstract of tripterygium wilfordii T7, on the human spermatozoa ATPase activity. METHODS: The specific activity of total ATPase and Na-K ATPase of spermatozoa was assayed in this experiment. RESULTS: (1) The effect of gossypol on the human spermatozoa ATPase activity is inhibitory and concentration-dependent; and (2) The abstract of tripterygium wilfodii T7 has no effect on the human spermatozoa ATPase activity. CONCLUSIONS: The antifertility effects of gossypol and T7 are different.

Antispermatogenic and androgenic activities of various extracts of Hibiscus rosa sinesis in albino mice.

The benzene chloroform and alcoholic extracts of the flowers of H.r.sinensis were administered (i.p.) at two different dose levels of 125 and 250 mg/kg body weight to adult male albino mice for 20 days. The results have shown decrease in the spermatogenic elements of testis and epididymal sperm count. High content of testicular cholesterol may be due to lowered androgen synthesis. The increase in the weight of accessory reproductive organs indicates the androgenicity of the plant extract itself, which is proved in the present study by testing the benzene extract in immature mice.

Preliminary studies on the role of plasminogen activator in seminal plasma of human and rhesus monkey.

Two types of plasminogen activators (PA), tissue type (tPA) and urokinase type (uPA), were identified in the seminal plasma of both the human and the rhesus monkey. We studied the possible relationship between PA activities in the seminal plasma and the sperm counts and motility and demonstrated that: (i) PA activity in human seminal plasma from infertile patients was associated with immotile spermatozoa; (ii) the treatment of fertile men with testosterone enanthate (TE) to induce azoospermia was accompanied by an increase in seminal PA activity; (iii) when monomer T4 (isolated from multiglycosides of Tripterygium wilforddi) was administered to fertile male rhesus monkeys to induce azoospermia, PA activities in seminal plasma increased considerably; and (iv) immunocytochemistry studies showed that both uPA and PAI-1 antigens were localized on the surface of human spermatozoa, indicating that human spermatozoa were capable of binding uPA and PAI-1 through their receptors or forming a complex. These data demonstrate that seminal PA activity may be related to azoospermia, and possibly, to the fertilizing capability of spermatozoa in primates.

Prolactin injection, a new contraceptive method: experimental study.

"Prolactin injection" is presented as a new contraceptive method. The method was tested in dogs. The dogs in the test group were injected with prolactin (PRL) in a dose of 600 micrograms/kg of body weight weekly for 6 months. During this period, the testicles, semen, reproductive hormones, renal function, and serum sodium and potassium were examined periodically. Testicular biopsy was obtained after 3 and 6 months of PRL injection. These investigations were repeated during the 6 months following withdrawal of the drug. Sperm count decreased to azoospermia in 3 months after PRL administration with decrease of sperm motility and increase of abnormal forms. Testicular biopsy showed degenerated seminiferous tubules. Reproductive hormones, renal function, and serum sodium and potassium revealed insignificant change (P > 0.05). Dog mating during the period of PRL administration induced no pregnancy. After 3 months of drug withdrawal, the sperm count normalized and dog mating produced pregnancy; offsprings showed no anomalies. The study demonstrates that PRL administration has the potential to be developed as a reversible male contraceptive.

PIP: A medical researcher in Egypt compared data on 14 male mongrel dogs that were injected with a 600 mcg/kg of body weight dose of prolactin every week for 6 months with data on 14 other dogs that received a saline injection, to determine whether prolactin administration would be a possible safe, effective, convenient, and reversible male contraceptive method. Experimental dogs experienced a gradual decrease in sperm count beginning at the end of the 2nd week of prolactin injection. They achieved near azoospermia after 6 weeks of treatment. At the end of 3 months, all but 1 dog were azoospermic. By the end of 6 months, all these dogs were azoospermic. Sperm motility fell and abnormal spermatozoa increased by 2 weeks after prolactin injection. These changes continued. At the end of 3 months of treatment, degenerative changes had occurred in the seminiferous tubules. Reproductive hormones, renal function, and serum sodium and potassium did not change appreciably. Three months after prolactin injection treatment began, the semen of 9 experimental dogs did not impregnate any fertile female dog, even after 8 repeated inseminations. It impregnated all female dogs 3 months after drug withdrawal, however. The puppies exhibited no developmental abnormalities. These findings suggest that prolactin administration may be a possible reversible male contraceptive. Perhaps it could be developed to be administered orally.