Testosterone Concentrations In Rats Research Articles

Effects of in utero benzo[a]pyrene exposure on the testis of rats during puberty and the protective effect of atorvastatin.

Benzo[a]pyrene (BaP) is a contaminant that is generated in the environment through processes such as smoke, incomplete combustion of fossil fuels, vehicle exhaust emissions, entry into the body is through inhalation, and consumption of contaminated food. It is an omnipresent environmental pollutant with unavoidable exposure. BaP metabolites are observed in the male reproductive system, especially in the testes and epididymis of animals, and are responsible for reduced testicular and epididymal function. The protective effect of atorvastatin (ATV) on testicular damage was investigated previously. The aim of the present study was to investigate the protective effect of ATV on testicular toxicity induced by benzo[a]pyrene (BaP) during pregnancy in Wistar rats. This experimental laboratory study involved 40 adult rats, divided into seven groups and maintained under standard environmental conditions. The groups received different diets [control, corn oil, ATV (10 mg/kg), BaP (10 and 20 mg/kg), and ATV + BaP (10 and 20 mg/kg)] at gestation Days 7-16, orally. Male offspring were examined 10 weeks after birth. Testis and serum samples were collected, and testosterone level, malondialdehyde (MDA), and glutathione (GSH) were measured. Histological and immunohistochemical assays were performed under a light microscope. Statistical analysis was conducted using SPSS, with analysis of variance and Tukey tests to assess significant differences between groups. ATV significantly reduced MDA, a marker of lipid peroxidation and oxidative stress in rat testes following BaP administration. Treatment with ATV at doses of 10 mg/kg increased GSH levels, correcting disruptions in the antioxidant system caused by BaP. Testosterone concentration in rats treated with ATV and BaP substantially prevented the decrease induced by BaP. Histomorphometry revealed that ATV significantly prevented the detrimental effects of BaP on the thickness of spermatogenic epithelium and the diameter of seminiferous tubules. Under ATV treatment, testicular tissue histopathology improved, and spermatogenesis returned to a almost back to normal state. Caspase-3 expression decreased, and apoptosis activity in testicular tissue improved under ATV treatment, indicating a positive effect of ATV in reducing apoptotic damage caused by BaP. In conclusion, exposure to BaP can induce oxidative stress-related damage to testicular tissue, as evidenced by an increase in MDA levels, which ATV treatment can mitigate. Additionally, ATV enhances intracellular antioxidant GSH and protects the testes against BaP-induced damage while increasing testosterone levels, which are reduced due to exposure to BaP.

The Effects of Administration of Vitamin D, Infliximab, and Leflunomide on Testosterone Concentrations in Rats under Atorvastatin Therapy.

Statins inhibit the 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase enzyme and thus reduce plasma cholesterol levels. Although decreased cholesterol level is the main target of anti-lipidemic drugs, cholesterol has an important role in the synthesis of lipid-based hormones such as testosterone. In this study, the alterations in serum testosterone levels were examined in rats under atorvastatin therapy and their responses to vitamin D, infliximab, and leflunomide supplementation were evaluated. Wistar rats were treated with atorvastatin (100 mg/kg) for 21 days to induce inhibition of the HMG-CoA reductase enzyme activity. Following statin therapy, rats received vitamin D (0.2 μg/kg/day) orally for 15 days, infliximab (7 mg/kg/day) intraperitoneally in two doses, or leflunomide (10 mg/kg/day) orally in two doses. Subsequently, the alterations in serum testosterone levels were measured by ELISA. Atorvastatin led to a decrease in the testosterone level compared to the vehicle group. Administration of vitamin D, infliximab, and leflunomide under HMG-CoA inhibition insignificantly increased the testosterone level compared to the atorvastatin control group. Furthermore, it appears that rats under statin administration respond better to treatment with leflunomide by achieving a greater induction in testosterone levels than with vitamin D or infliximab. Our data provide evidence that administration of vitamin D, infliximab, and leflunomide in rats under atorvastatin treatment may ameliorate the serum testosterone levels.

Ectopic Organogenesis after Allotransplantation of Freshly Removed or Cryopreserved Neonatal Testicle under the Renal Capsule in Rats.

In experiments on white outbred male rats, a freshly removed (20 experiments) or cryopreserved (10 experiments) testicle from newborn rats (1-2 days after birth) was transplanted under the renal capsule after bilateral orchiectomy. In all experiments with transplantation of freshly removed testicle, it was engrafted. In 3 months, histological examination revealed the formation of mature seminiferous tubules, but spermatogenesis was blocked at the stage of spermatogonia; groups of proliferating Leydig cells in the loose connective tissue between the tubules were also seen. In 6 and 12 months, the status of the seminiferous tubules remained unchanged, but structures typical of the epididymis and developing vas deferens were revealed. The number of proliferating Leydig cells increased. The initially low testosterone concentration in the blood of castrated males increased significantly as soon as in 1 month after transplantation and grew up to 3 months, remaining at a level ~50% of normal. Engraftment of cryopreserved neonatal testicular tissue was observed in 60% cases, however, engrafted tissue, similar to the fresh one, retained the ability for organogenesis with the formation of mature seminiferous tubules, epididymis, and groups of proliferating Leydig cells. The dynamics of blood testosterone concentration in rats with cryopreserved and fresh transplantation was similar. Subcapsular transplantation did not adversely affect the kidneys, which was seen from normal histological structure of the kidneys and creatinine and urea concentrations in the blood.

Simultaneous Effect of Kisspeptin and Galanin on Serum Luteinizing Hormone and Testosterone Levels in Male Rats

Background: Galanin is a neuropeptide with a wide range of physiological functions that also has stimulatory effects on the reproductive axis. Kisspeptin is a crucial neuropeptide for the stimulation of the reproductive function. In the present study, the interaction of kisspeptin and the galanin signaling system was investigated on the mean serum luteinizing hormone (LH) and the testosterone concentrations in rats. Materials and Methods: Fifty-five male Wistar rats in 11 groups (n=5 per group) received saline, kisspeptin (1nmol), P234(1nmol), galanin(1nmol), galantide (1nmol) or simultaneous injections of them via third cerebral ventricle at 07:00 - 07:30. Blood samples were collected at 30 min following the injections. Hormone concentrations were measured using rat kit and the method of the radio-immunoassay (RIA). Results: Kisspeptin or galanin injection significantly increased both the mean serum LH and the testosterone concentration compared to saline (P<0.05). The co-administration of kisspeptin/galanin increased the mean serum LH and the testosterone concentration significantly compared to galanin or saline (P<0.05). The co-administration of kisspeptin/galanin decreased the mean serum LH concentration compared to kisspeptin, this reduction, however, was not statistically significant. Also, testosterone concentration declined in the kisspeptin/galanin group compared to kisspeptin group. Galantide or p234 injection decreases the mean serum LH and the testosterone concentration compared to galanin and kisspeptin, respectively. The co-administration of galantide/p234 lowers the mean serum LH concentration compared to saline. Conclusion: The interaction of hypothalamic galanin and kisspeptin signaling pathways may play an important role in the modulation of hormonal control of the hypothalamus-pituitary-gonadal axis. [GMJ. 2017;6(1):23-29]

Restraint stress exacerbates alcohol-induced reproductive toxicity in male rats

Cumulative exposure to multiple stresses may lead to aggravating the toxicity of each stress, qualitatively or quantitatively altering biological responses because of toxicological interaction. In this study, we intended to determine the possible effects of restraint stress on reproductive toxicity due to ethanol usage in male rats. Early pubertal male Wistar rats were subjected to either restraint stress (5 h/day) or alcohol intoxication (2 mg/kg body weight) or both for 60 days. Body weights of control and experimental rats were similar during the 60 days of this study. Testes were harvested, weighed, and prepared for enzyme assays, and cauda epididymides were isolated for the determination of density, motility, and viability of stored spermatozoa. Restraint stress or alcohol treatment significantly reduced testis weight and caused significant reductions in steroidogenesis and spermatogenesis. Mean density, motility, and viability of stored spermatozoa were reduced in experimental rats. Plasma testosterone concentrations in rats subjected to restraint stress or alcohol were decreased compared with those of controls, concomitant with increased concentrations of LH and FSH in experimental rats. These data suggest that sub-chronic exposure to restraint stress or alcohol contribute to reduce testicular and epididymal function in exposed rats. The study also suggests that restraint stress exacerbates alcohol-induced reproductive toxicity in rats.

Effect of Cordyceps militaris on Testosterone Production in Sprague-Dawley Rats

Some of men have been suffered from the insufficient secretion of testosterone causing by physical factors, social and psychological factors. Testosterone is an essential steroid hormone controlling male reproductive function. Alternative medicines in plants, fungi, and insects have been studied to enhance sexuality. <TEX>$Cordyceps$</TEX> species including <TEX>$Cordyceps$</TEX> <TEX>$sinensis$</TEX> (CS) and <TEX>$C.$</TEX> <TEX>$militaris$</TEX>(CM) has been used as for the enhancement of sexual functionfor hundreds of years in Far East Asian. In the present study, we determined the effect of fruiting bodies of <TEX>$C.$</TEX> <TEX>$militaris$</TEX> which cultured on bee drone medium (CMD) and brown rice medium (CMB) on testosterone concentration in Sprague-Dawley rats. Eighteen rats per group were housed to regular diet or diet supplemented with CMB and CDD, respectively for 4 weeks. Serum was collected from 6 rats per group. Results showed that changes of the body weight, food and water intake of the rats were not observed in this study. However, both CMB and CDD increased the serum testosterone concentration in rats. Furthermore, CMD significantly stimulated testosterone production (p <0.05) compared to the control. Hence, it suggests that <TEX>$C.$</TEX> <TEX>$militaris$</TEX> fruiting bodymight be developed as a complementary medicine to improve sexual hormones.

Calorie restriction at increasing levels leads to augmented concentrations of corticosterone and decreasing concentrations of testosterone in rats

The influence of calorie restriction (CR) on increasing life span, enhancing immunocompetence, and reducing the incidence of age-related diseases is well established. Evidence points to the involvement of neuroendocrine alterations in these beneficial effects. Accordingly, we hypothesized that CR will result in significant alterations to the hormones investigated. Little attention has been directed toward ascertaining the doses of CR required to obtain such alterations and, indeed, whether a dose-response exists. Adult rats were subjected to 1 of 5 dietary regimens: control, CR12.5%, CR25%, CR37.5%, or CR50%. Rats were decapitated 3 weeks following the onset of restriction; and trunk blood was collected and assayed for concentrations of serum adrenocorticotropic hormone, corticosterone, and testosterone, as well as plasma concentrations of noradrenalin and adrenalin. No effect was found as a result of dietary manipulation for serum concentrations of adrenocorticotropic hormone. However, all doses of CR resulted in increased serum corticosterone in a dose-response trend. A dose-response was also observed for serum testosterone, with higher doses of CR associated with lower testosterone. Concentrations of noradrenalin were not found to be altered by any CR dose, although a trend toward a down-regulation at CR50% was observed. Plasma adrenalin displayed a biphasic distribution with reductions observed at CR25% and CR50%, although the down-regulations only attained statistical significance relative to the CR37.5% and not the control group. As well as reporting the effect of CR on multiple hormones within individual animals, these results go some way in determining the optimal levels of CR needed to induce neuroendocrinologic alterations.

Inhibitory Effects of Oral Administration of Nonylphenol on Testosterone Production in Rat Leydig Cells.

Nonylphenol (NP) is the final metabolite of nonyphenol polyethoxylate (NPE), a non-ionic surfactant that is frenquently incorporated into detergent and pesticide formulation. This metabolite has a structure mimicking 17β-estradiol and has been reported to have xenoestrogenic effects, and may affect the endocrine system. Its effects on testosterone release from Leydig cells are unclear. In the present study, we investigated the influence of NP, administered orally (gavage) or delivered via spray on Leydig cell activity and testosterone release. Rat Leydig cells were prepared by enzymatic dispersion and then percoll density gradient centrifugation. The purified Leydig cells (1×105 cells/ml) were incubated with or without human chorionic gonadotropin (hCG, 0.05 IU/ml), forskolin (FSK, an adenylyl cyclase activator, 10-5 M), A23187 (a calcium ionophore, 10-5 M), androstenedione (a precursor of testosterone, 10-6 M), or 8-bromo-adenosine-3',5'-cyclic monophosphate (8-Br-cAMP, a cell-permeable cAMP analogue, 10-5 M) at 34°C for 1 hour. Testosterone levels in the media were measured by radioimmunoassay. The data showed that the administration of NP via spray did not affect steroidogenesis of testosterone in rat Leydig cells. In contrast, oral administration of NP (100 mg/kg) once per day for 30 days had inhibitory effect on steroidogenesis of testosterone in rat Leydig cells. In the in vivo experiments, administration of NP by oral ingestion decreased plasma testosterone concentrations in rats. Furthermore, oral NP induced a significant inhibition on the release of testosterone in vitro in response to hCG, 8-Br-cAMP, and FSK. Moreover, administration of NP also caused a decrease on testosterone release in response to androstenedione and A23187. The in vitro experiments revealed that oral NP at a dose of 100 mg/kg/day suppressed testosterone biosynthesis by inhibiting the 17β-hydroxysteroid dehydrogenase activity, cyclic AMP pathway and calcium influx in Leydig cells. These results suggested that oral administration of NP had inhibitory effects on steroidogenesis of testosterone in rat Leydig cells and might provide more information about the toxic effects of nonylphenol on reproductive endocrine disrupt.

Increased serum and testicular androgen levels in F1 rats with lifetime exposure to soy isoflavones

The consequences of dietary soy isoflavones on serum and testicular androgen levels were examined in F1 male rats from a multigeneration study investigating the effects of diets varying in isoflavone content. Rats were fed either a soy-free casein based diet (AIN93G) or a diet in which alcohol-washed soy protein replaced casein as the protein source and to which increasing amounts of Novasoy, a commercially available isoflavone supplement were added. Analysis of these diets showed that the isoflavone content in each diet was 0 (diet 1; casein based control), 31.7 (diet 2; alcohol-washed soy-based diet control), 36.1 (diet 3), 74.5 (diet 4), 235.6 (diet 5) and 1046.6 (diet 6) mg total isoflavones/kg pelleted diet. The levels of isoflavones in diet 1 would represent a daily intake level of 0 mg isoflavones, diets 2 and 3 estimate a low soy-containing human diet (e.g. North American), diet 4 would correspond to Asian diets (e.g. Japanese) or adult humans taking isoflavone supplements, diet 5 approximates the isoflavone intake by babies fed soy based infant formula and diet 6 approximates fivefold the intake levels by babies or 10-fold the intake levels of adults consuming high isoflavone containing diets. Serum testosterone (T) from F1 male rats sacrificed on postnatal days (PND) 28, 70, 120, 240 and 360 were low at PND 28 (0.4 ng/ml), increased approximately five to sixfold at PND 70 (2.5–3.0 ng/ml) and thereafter declined to a steady state level of ∼1 ng/ml by PND 120. However, rats on diets 5 and 6 demonstrated altered serum testosterone profiles such that at days 120, testosterone levels remained significantly elevated at ∼3 ng/ml ( P<0.05). Serum dihydrotestosterone levels exhibited similar profiles and the levels in PND 120 rats on diet 5 or 6 were also significantly elevated (two to threefold, P<0.05). The intra-testicular testosterone concentration in rats on diet 5 was also elevated at PND 120 compared with diet 1 ( P<0.05). These findings show that F1 male rats continuously exposed to a mixture of dietary soy isoflavones from conception onwards exhibit altered serum and testicular androgen profiles.

Synthesis and evaluation of 17-aliphatic heterocycle-substituted steroidal inhibitors of 17alpha-hydroxylase/C17-20-lyase (P450 17).

In the search for potent inhibitors of P450 17, the key enzyme in androgen biosynthesis, a series of steroidal inhibitors were synthesized and tested toward rat and human P450 17. Small aliphatic heterocycles (aziridine, oxirane, thiirane, diaziridine, diazirine, azetidine) were introduced into the 17beta-position of anstrost-5-en-3beta-ol. After identifying that aziridine is the most suitable functional group to coordinate with the heme iron, modifications of the steroidal skeleton were performed for further optimization. A wide range of inhibitory potencies toward P450 17 were found for the 21 test compounds. The most potent inhibitors toward the human and rat enzyme were aziridine compounds 3 (IC(50) rat: 0.21 microM, K(i) = 3 nM; IC(50) human: 0.54 microM, K(i) = 8 nM), 5 (IC(50) rat: 0.43 microM, K(i) = 7 nM; IC(50) human: 0.29 microM, K(i) = 4 nM), and 8 (21R:21S = 1:1; IC(50) rat: 0.53 microM, K(i) = 9 nM; IC(50) human: 0.40 microM, K(i) = 6 nM) which were more potent than the reference ketoconazole (IC(50) rat: 67 microM; IC(50) human: 0.74 microM). The inhibitory potency depends markedly on the stereochemistry at C20 of the inhibitors. This effect is more pronounced for the rat enzyme. Tested for selectivity, the highly potent inhibitors show poor inhibitory activity toward P450 arom, P450 scc, P450 TxA(2), and 5alpha-reductase. Tested for in vivo activity, 3 and 8 (0.019 mmol/kg) decreased the plasma testosterone concentration in rats by 81% and 84% after 2 h.

Synthesis and Evaluation of Novel Steroidal Oxime Inhibitors of P450 17 (17α-Hydroxylase/C17−20-Lyase) and 5α-Reductase Types 1 and 2

17 alpha-Hydroxylase/C17-20-lyase (P450 17, CYP 17) and 5 alpha-reductase are the key enzymes in androgen biosynthesis and targets for the treatment of prostate cancer and benign prostatic hyperplasia. In the search of inhibitors for both enzymes, 23 pregnenolone- or progesterone-based steroids were synthesized bearing an oxime group connected directly or via a spacer to the steroidal D-ring. Tested for inhibition of human and rat P450 17, some pregnenolone (9, 11, 14) and a series of progesterone compounds (17-20) turned out to be highly active inhibitors of the human enzyme. The most active compound was Z-21-hydroxyiminopregna-5, 17(20)-dien-3 beta-ol (9) showing K(i) values of 44 and 3.4 nM for the human and rat enzymes, respectively, and a type II UV-difference spectrum indicating a coordinate bond between the oxime group and the heme iron. In contrast to the pregnenolones which showed no inhibition of 5 alpha-reductase isozymes 1 and 2, the progesterones 16, 17, 20, 21, and 23 showed marked inhibition, especially toward the type 2 enzyme. Compounds 17 and 20 were identified as potent dual inhibitors of both P450 17 and 5 alpha-reductase. Tested for selectivity, the most potent P450 17 inhibitors 9, 10, and 14 showed no or only marginal inhibition of P450 arom, P450 scc, and P450 TxA(2). Selected compounds were tested for inhibition of the target enzymes using whole-cell assays. Compounds 9-11 strongly inhibited P450 17 being coexpressed with NADPH-P450 reductase in E. coli cells, and 16, 20, and 23 markedly inhibited 5 alpha-reductase expressed in HEK 293 cells. Tested for in vivo activity, 9 (0.019 mmol/kg) decreased the plasma testosterone concentration in rats after 2 and 6 h by 57% and 44%.

Circularly polarized, sinusoidal, 50 Hz magnetic field exposure does not influence plasma testosterone levels of rats

We exposed rats to circularly polarized 50 Hz magnetic fields to determine if plasma testosterone concentration was affected. Previous experiments indicate that magnetic fields suppress the nighttime rise in melatonin, suggesting that other neuroendocrine changes might occur as well. Male Wistar-King rats were exposed almost continuously for 6 weeks to magnetic flux densities of 1, 5, or 50 microT. Blood samples were obtained by decapitation at 12:00 h and 24:00 h. Plasma testosterone concentration showed a significant day-night difference, with a higher level at 12:00 h when studied in July and December, but night difference, with a higher level at 12:00 h when studied in July and December, but the day-night difference disappeared when concentrations were studied in April. In three experiments, magnetic field exposure had no statistically significant effect on plasma testosterone levels compared with the sham-exposed groups. These findings indicate that 6 weeks of nearly continuous exposure to circularly polarized, 50 Hz magnetic fields did not change plasma testosterone concentration in rats.

Inhibition of testosterone synthesis by ethanol: Role of luteinizing hormone

The effects of acute ethanol intake (1.5 g/kg) on plasma testosterone and luteinizing hormone (LH) concentrations were examined in male Long-Evans rats. Ethanol decreased the serum LH concentrations by 21% and 42% 30 and 60 minutes after ethanol administration respectively. The testosterone concentrations decreased later (30 min: +8%; 60 min; −30%). The LH concentrations were highly correlated with subsequent (60 min later) testosterone concentrations (LH 30min: r = .688, p<0.001, n = 25; LH 60 min: r = .678, p<0.001), but less so with concurrent testosterone concentrations (30 min: r = .187, N.S.; 60 min: r = .552, p<0.004). To further test the influence of LH, naloxone (11 mg/kg) was administered, which elevated the LH levels within 30 min by 103% in controls. Naloxone also increased serum LH concentration by 34% in ethanol-treated rats at 30 min, but these animals nevertheless had lower ( p<0.01) testosterone levels at 60 min than did control animals without naloxone and ethanol treatment. It is concluded that although ethanol-induced changes in serum LH levels may play a role in the decrease of serum testosterone concentrations in rats, there are also other mechanisms by which ethanol may produce these effects.

Inhibition of testosterone biosynthesis by ethanol. Relation to hepatic and testicular acetaldehyde, ketone bodies and cytosolic redox state in rats.

In experiments in which liver and testis freeze-stops were performed on pentobarbital-anaesthetized rats, ethanol (1.5 g/kg body wt.) reduced plasma testosterone concentration from 13.1 to 3.2 nmol/litre. 4-Methylpyrazole abolished the ethanol-induced hepatic and testicular increase in the lactate/pyruvate ratio, and the testicular acetaldehyde level, but did not diminish the reduction in plasma testosterone concentration. In testes, but not in liver, ethanol decreased the 3-hydroxybutyrate/acetoacetate ratio, and 4-methylpyrazole did not prevent this effect. In experiments in which freeze-stop was performed after cervical dislocation, ethanol decreased the testis testosterone concentration from 590 to 220 pmol per g wet wt. The effects of ethanol and 4-methylpyrazole on testis acetaldehyde, lactate/pyruvate and 3-hydroxybutyrate/acetoacetate ratios were the same as found during anaesthesia. The NAD+-dependent ethanol oxidation capacity in testis ranged from 0.1 to 0.2 mumol/min per g wet wt. and seemed to be inhibited by 4-methylpyrazole both in vivo and in vitro. In additional experiments, ethanol doses between 0.3 and 0.9 g/kg body wt. did not alter the plasma testosterone concentration in rats treated, or not treated, with cyanamide, which induced elevated acetaldehyde levels in blood and testes. The results suggest that ethanol-induced inhibition of testosterone biosynthesis was not caused by extratesticular redox increases, or by extra- or intra-testicular acetaldehyde per se. The inhibition is accompanied by changes in testicular ketone-body metabolism.