Influence of genistein aglycone on some male reproductive functions in pubertal Holtzman rats

Background: Genistein, a phytoestrogen found in abundance in soya has been known to adversely influence male reproductive system. The effects of genistein on some male reproductive functions were investigated in pubertal laboratory rats. Methods: Male Holtzman rats, 70-75 days old and weighing 200-250g were used for the study. They were grouped into five groups of ten rats each. Group 1 (control) received distilled water. Groups 2, 3, 4, and 5 were administered orally with 0.5, 1, 2, 4 mg/ kg body weight of genistein respectively for a period of 60 days. Daily feed consumption and final body weight were recorded. Sperm count and motility were analysed along with the serum testosterone, estradiol and leptin levels. Acrosome reaction (AcR) was assessed using PSA-FITC with calcium and progesterone as stimulants. Male potency and fertility index were also calculated. Results: There was significant decrease in feed consumption in the 2 and 4 mg/kg genistein groups within the first 15 days of the experiment with a corresponding decrease in final body weights in 0.5, 2 and 4 mg/kg groups. A significant decrease was recorded in the right and the left absolute testicular weights in 0.5, 2 and 4 mg/kg groups while the right and left absolute epididymal weight and the prostate gland were significantly reduced in 2 and 4 mg/kg groups. The result showed a significant decrease and increase in serum testosterone level in 0.5 and 1 mg/kg groups respectively. Estradiol level was significantly reduced in all the genistein treated groups. Serum leptin was significantly increased in 1 mg/kg group. Sperm count was significantly reduced in 4 mg/kg group while sperm motility was reduced in the 2, and 4 mg/kg groups. Sperm track speed; lateral amplitude and elongation were significantly increased in all the genistein groups. Sperm path velocity was increased in all genistein groups except in the 4 mg/kg group. Progesterone stimulated AcR resulted in largely intact acrosome in all genistein group while calcium significantly increased percentage of reacted acrosome in 0.5 mg/kg group. Combined stimulation with progesterone and calcium increased AcR when compared with either of the stimulators alone. Potency and fertility index were significantly reduced in both 2 and 4 mg/kg groups while days of cohabitation before successful mating were increased in the 2 and 4 mg/kg groups. Ploidy analysis showed a significant increase and decrease in the population of elongating spermatid (HCI) and round spermatid (IC) in 2 and 4 mg/kg respectively. Pre-leptotene spermatocytes and primary spermatozoa population were significantly decreased in 4 mg/kg group. Conclusion: Oral administration of genistein to pubertal Holtzman male rats adversely influenced some important male reproductive functions

The metabolism of some steroids in the horse.

This thesis has been mainly concerned with the determination of the overall urinary excretion and the characterization of the neutral urinary metabolites of dexamethasone, testosterone, 1-dehydrotestosterone and 19-nortestosterone after intramuscular administration to castrated horses (ponies and thoroughbreds). The percentages of the free and the conjugated steroids were determined after solvent extraction and Sephadex LH-20 column chromatography. The urinary steroids were extracted by Amberlite XAD-2 and separated into glucuronides and sulphoconjugates by Sephadex LH-20 chromatography. After enzyme hydrolysis and solvolysis respectively, the freed steroids were fractionated on Kieselgel H columns and after derivatization as their trimethylsilyl ethers or methoxime-trimethylsilyl ethers, were analysed by combined gas chromatography-mass spectrometry. The metabolites of dexamethasone were identified by microchemical reactions and thin-layer chromatography. Results of the analysis have shown that in general the pathways of biotransformation (Phase I and Phase II) of these steroids in the equine castrate were qualitatively similar to those previously reported for other species. However, the quantitative differences in the excretion and relative importance of these pathways are attributed to the effects of species difference and the structural modification of the steroids. Epimerization at C-17 with stereoselective conjugation to glucuronic acid, stereo-selective reduction of the Delta4-3-oxo group with the formation of 3beta,5alpha-isomer, stereoselective conjugation of the 17beta-hydroxyl group to sulphuric acid and the hydroxylation at C-16 were major transformations observed for the anabolic steroids. 6-Hydroxylation and reduction of the 20-ketone were the major transformations for dexamethasone. On the basis of the results obtained from this work it is now possible to develop specific gc-ms methods to confirm the administration of these steroids to horses

The metabolism of some steroids in the horse.

This thesis has been mainly concerned with the determination of the overall urinary excretion and the characterization of the neutral urinary metabolites of dexamethasone, testosterone, 1-dehydrotestosterone and 19-nortestosterone after intramuscular administration to castrated horses (ponies and thoroughbreds). The percentages of the free and the conjugated steroids were determined after solvent extraction and Sephadex LH-20 column chromatography. The urinary steroids were extracted by Amberlite XAD-2 and separated into glucuronides and sulphoconjugates by Sephadex LH-20 chromatography. After enzyme hydrolysis and solvolysis respectively, the freed steroids were fractionated on Kieselgel H columns and after derivatization as their trimethylsilyl ethers or methoxime-trimethylsilyl ethers, were analysed by combined gas chromatography-mass spectrometry. The metabolites of dexamethasone were identified by microchemical reactions and thin-layer chromatography. Results of the analysis have shown that in general the pathways of biotransformation (Phase I and Phase II) of these steroids in the equine castrate were qualitatively similar to those previously reported for other species. However, the quantitative differences in the excretion and relative importance of these pathways are attributed to the effects of species difference and the structural modification of the steroids. Epimerization at C-17 with stereoselective conjugation to glucuronic acid, stereo-selective reduction of the Delta4-3-oxo group with the formation of 3beta,5alpha-isomer, stereoselective conjugation of the 17beta-hydroxyl group to sulphuric acid and the hydroxylation at C-16 were major transformations observed for the anabolic steroids. 6-Hydroxylation and reduction of the 20-ketone were the major transformations for dexamethasone. On the basis of the results obtained from this work it is now possible to develop specific gc-ms methods to confirm the administration of these steroids to horses