Testicular Testosterone Concentrations Research Articles

26 Trace minerals to improve bull fertility

Abstract Bull fertility is paramount for reproductive success in cow-calf production systems and there are a variety of factors that influence bull fertility. Trace mineral requirements and recommendations have been established for growing and finishing steers, as well as lactating and pregnant cows, yet no specific requirements have been established for young or mature beef bulls. Nonetheless, trace minerals have been shown to have an important role in sexual maturation and semen quality of ruminants. Research performed in rams indicates that zinc deficiency results in reduced testicular development, decreased testicular concentrations of testosterone, and impaired development of seminiferous tubules. Deficiency of Zn has also been reported to negatively impact male fertility in humans, where men diagnosed with infertility responded positively to Zn supplementation. In growing bulls, diets with greater concentrations of organic forms of Zn, Cu, and Mn, tended to result in earlier attainment of puberty compared with control diets containing inorganic minerals. Furthermore, greater liver concentrations of Zn have been negatively correlated with sperm acrosomal damage in growing bulls, based on flow cytometry analyses. More recent research indicates that the use of hydroxichloride-based trace minerals results in greater liver concentrations of Zn and Cu compared with sulfate sources. While evidence indicates that trace mineral status has an important role in bull fertility, further research is required to better understand the contributions of trace minerals to semen quality. In addition, investigating the impact of trace mineral sources with greater bioavailability on semen quality may result in the development of practical recommendations that optimize bull fertility.

P-002 Acetate attenuates cyclophosphamide-induced testicular toxicity by modulating steroidogenic and apoptotic signaling via an oxidative stress-dependent pathway

Abstract Study question Will acetate therapy attenuate cyclophosphamide-induced testicular toxicity when used concomitantly? Summary answer Acetate attenuated cyclophosphamide-induced testicular toxicity by modulating steroidogenic and apoptotic signaling via an oxidative stress-dependent pathway. What is known already Cyclophosphamide is an anthracycline that is effective in the management of cancers. Nonetheless, a major drawback of this drug is its testicular toxicity. Available evidence in the literature revealed that cyclophosphamide induces testicular toxicity by targeting steroidogenesis and activating oxidative stress, inflammation, and apoptosis. On the other hand, acetate has been reported to exert antioxidant, anti-inflammatory, and anti-apoptotic activities. However, there is no report on the effect of acetate on cyclophosphamide-induced testicular toxicity. Study design, size, duration This is a prospective experimental study using animal model. Twenty-four sexually mature litter-mate male Wistar rats of comparable weight were used for the study. The study lasted 2 weeks. Participants/materials, setting, methods Animals were acclimatized for two weeks, and then randomized into four groups. The control rats received 0.5mL of distilled water p.o for 14 days, acetate-treated rats received 200 mg/kg/day of acetate p.o for 14 days, cyclophosphamide-treated rats received 150 mg/kg of cyclophosphamide i.p on day 8, while acetate + cyclophosphamide-treated rats received treatment as acetate-treated and cyclophosphamide-treated. The doses of drugs used were the Human Equivalent doses for rats and as earlier reported. Main results and the role of chance Acetate attenuated cyclophosphamide-induced alterations in testicular histoarchitecture and the rise in testicular activities of gamma glutamyl transferase and lactate dehydrogenase activities, and lactate levels. In addition, acetate abrogated cyclophosphamide-induced rise in testicular malondialdehyde, TNF-α, IL-1β, NF-kB, and Bax levels, and myeloperoxidase, caspase 9, and caspase 3 activities, which was accompanied by acetate-driven blockade of cyclophosphamide-induced decline in reduced glutathione, Nrf2, and Bcl-2 levels, and glutathione peroxidase, glutathione-S-transferase, superoxide dismutase and catalase activities. Furthermore, acetate alleviated cyclophosphamide-induced suppression of circulatory levels of FSH, LH, and testosterone, testicular concentrations of 3β-HSD, 17β-HSD, and testosterone, and spermatogenesis and lowered sperm quality. Limitations, reasons for caution This was an experimental study using a rat model; hence, the results of this study should be extrapolated to humans with caution. Clinical trials are recommended to validate these findings. Wider implications of the findings The present study revealed the protective effect of acetate treatment on cyclophosphamide-induced testicular toxicity. These findings provide convincing evidence of the attenuating effect of acetate in cyclophosphamide-induced testicular toxicity. Trial registration number N/A

Ameliorative impacts of propolis against testicular toxicity promoted by doxorubicin

Background and Aim: Doxorubicin (DOX) is often used as a chemotherapeutic agent, although it may damage testicular functions. This study was designed to investigate the protective effects of propolis on testicular histological changes, semen parameters, and testosterone concentrations as a means of protecting against testicular damage caused by DOX chemotherapy. Materials and Methods: Forty-eight male Wistar rats were divided into four groups with 12 animals per group. The first group served as the control. Rats in the second group were administered 4 mg/kg DOX. The third group was administered 4 mg/kg of DOX and 30 mg/kg b.w. propolis. The fourth group was orally dosed daily with 30 mg/kg b.w. propolis. Results: DOX treatment resulted in a significantly decreased weight gain (WG) rate compared with the control, whereas DOX + propolis resulted in improved WG and returned to the normal range. Testosterone levels were comparable among the experimental groups, with a significant increase in the propolis-treated group. In addition, DOX-treated groups exhibited a remarkable depletion in sperm counts, motility, and viability compared to the other groups. Conclusion: Most of the histological and hormonal changes resulting from the toxicity of DOX returned to almost normal after treatment of rats with the aqueous extract of propolis, indicating that propolis ameliorated the effects of DOX poisoning on testicular function in male rats. Keywords: doxorubicin, histopathology, propolis, testicular toxicity.

L-Arginine abrogates maternal and pre-pubertal codeine exposure-induced impaired spermatogenesis and sperm quality by modulating the levels of mRNA encoding spermatogenic genes.

Although, codeine has been demonstrated to lower sperm quality; the effects of maternal and prepubertal codeine exposure on male offspring is yet to be reported. In addition, the effect of arginine on codeine-induced decline in sperm quality has not been explored. This study investigated the impact of maternal and prepubertal codeine exposure on spermatogenesis and sperm quality in F1 male Wistar rats to study the effect that codeine may have during recreational use in humans. Also, the effect of arginine supplementation on codeine-induced alteration in spermatogenesis and sperm quality was evaluated. Female rats were treated with either 0.5 ml distilled water or codeine orally for eight weeks, and then mated with male rats (female:male, 2:1). The F1 male offsprings of both cohorts were weaned at 3 weeks old and administered distilled water, codeine, arginine, or codeine with arginine orally for eight weeks. Prepubertal codeine exposure in rats whose dams (female parents) were exposed to codeine delayed puberty and reduced the weight at puberty. Prepubertal codeine exposure exacerbated maternal codeine exposure-induced reduced total and daily spermatid production, sperm count, sperm motility, and normal sperm form, as well as impaired sperm plasma membrane integrity and increased not intact acrosome and damaged sperm DNA integrity. These perturbations were accompanied by a decrease in mRNA levels encoding spermatogenic genes, testicular testosterone and androgen receptor (AR) concentrations, and upregulation of sperm 8-hydroxydeoxyguanosine (8OHdG). Prepubertal arginine supplementation mitigated codeine-induced alterations. This study provides novel experimental evidence that maternal and prepubertal codeine exposure reprogramed spermatogenesis and sperm quality of male FI generation by decreasing mRNA levels encoding spermatogenic genes and AR via oxidative stress-mediated signaling, which was abrogated by prepubertal arginine supplementation.

Mucuna pruriens seed repeals the hypothalamic-pituitary-testicular axis disruption following carbamazepine treatment in male Wistar rats.

This study examined the potential effects of Mucuna pruriens (MP) seed powder on the disruptions of the hypothalamic-pituitary-testicular axis caused by the carbamazepine (CBZ) treatment in male Wistar rats. A total of 35 male Wistar rats were randomized into 5 groups (n=7). The animal in group 1 received normal saline (0.2 ml) orally, while groups 2-5 received carbamazepine (CBZ) 25 mg/kg per oral. Groups 1, and 2 were fed with standard rats' chow, while groups 3-5 rats were supplied with a diet containing MP seed powder at 2.25 g, 1.5 g, and 0.75 g respectively. The serum level of male reproductive hormones, estradiol, seminal profiles, and histoarchitecture of the hypothalamus, pituitary, and testis was delineated. Descriptive and inferential statistics were used to analyze the result. There was a marked decrease in the testicular weight, follicle-stimulating hormone, testosterone concentration, and normal sperm cells in the CBZ, and CBZ + MP (2.25 mg/kg) treatment groups. There was a marked increase in the testicular tissue lipid peroxidation in the CBZ, and CBZ + MP (g) treated rats in addition to various morphological alterations in the hypothalamus, pituitary, and testis. These anomalies were receded in the CBZ + MP (1.5 g), and CBZ + MP (0.75 g) treatment groups. Consumption of MP (1.5 g, and 0.75 g) may alleviate the injurious effects of CBZ treatment on the hypothalamic-pituitary-testicular functions.

P-008 Acetate ameliorates doxorubicin-induced testicular toxicity by modulating Nrf2/NFkB pathway and apoptotic signaling

Abstract Study question Will acetate treatment ameliorate doxorubicin-induced testicular toxicity when used concomitantly? Summary answer Acetate ameliorated doxorubicin-induced testicular toxicity by modulating Nrf2/NFkB and apoptotic signaling. What is known already Doxorubicin is an antineoplastic agent that is effective in the management of several forms of cancers. However, a major limitation of this drug is its testicular toxicity. Studies have revealed that doxorubicin impairs male fertility by inducing testicular injury via downregulation of Nrf2 and upregulation of NFkB and apoptotic signaling. On the other hand, acetate upregulates Nrf2 and downregulates NFkB and apoptotic signaling. Yet, no study has reported the impact of acetate on doxorubicin-induced testicular toxicity. Study design, size, duration This is a prospective experimental study using animal model. Thirty two sexually mature litter-mate male Wistar rats of comparable weight were used for the study. The study lasted 8 weeks. Participants/materials, setting, methods Animals were acclimatized for two weeks, then randomized into four groups (n = 8). The control rats received 0.5mL of distilled water as vehicle p.o for 14 days, acetate-treated rats received 200 mg/kg/day of acetate p.o for 14 days, doxorubicin-treated rats received 7 mg/kg of doxorubicin i.p on day 8, while acetate + doxorubicin-treated rats received treatment as acetate-treated and doxorubicin-treated. The doses of drugs used were the Human Equivalent doses for rats. Main results and the role of chance Acetate ameliorated doxorubicin-induced rise in testicular activities of gamma glutamyl transferase, lactate dehydrogenase activity, and lactate levels. Also, acetate ameliorated doxorubicin-induced decline in the activities of testicular superoxide dismutase and catalase, and glutathione and Nrf2 concentrations. Acetate also alleviated doxorubicin-induced rise in testicular MPO activities, malondialdehyde, TNF-α, IL-1β, and NF-kB levels. More so, acetate ameliorated doxorubicin-induced reduction in spermatogenic indices and sperm quality, circulatory levels of FSH, LH, and testosterone, and testicular concentrations of 3β-HSD, 17β-HSD, and testosterone. These events were associated with dampening of doxorubicin-induced upregulation of Bax, caspase 3 and caspase 9 activities and doxorubicin-induced downregulation of BCl-2 by acetate. Limitations, reasons for caution This study was conducted in a rat model; therefore the results of the present study should be extrapolated to human with caution. Clinical trials are recommended to validate these findings. Wider implications of the findings The present study demonstrated the ameliorative effect of acetate treatment on doxorubicin-induced testicular toxicity. These findings provide substantive evidence of the possible protective role of acetate in doxorubicin-induced testicular toxicity. Trial registration number Not applicable

P38 mediates T-2 toxin-induced Leydig cell testosterone synthesis disorder

T-2 toxin is an unavoidable food and feed contaminant that seriously threatens human and animal health. Exposure to T-2 toxin can cause testosterone synthesis disorder in male animals, but the molecular mechanism is still not completely clear. The MAPK pathway participates in the regulation of testosterone synthesis by Leydig cells, but it is unclear whether the MAPK pathway participates in T-2 toxin-induced testosterone synthesis disorders. In this research, testosterone synthesis capacity, testosterone synthase expression and MAPK pathway activation were examined in male mice and TM3 cells exposed to T-2 toxin. The results showed that T-2 toxin exposure decreased testicular volume and caused pathological changes in the microstructure and ultrastructure of testicular Leydig cells. T-2 toxin exposure also decreased testicular testosterone content and the protein expression of testosterone synthase. In vitro, T-2 toxin inhibited cell viability and decreased the expression of testosterone synthase in TM3 cells, and it decreased the testosterone contents in cell culture supernatants. Moreover, T-2 toxin activated the MAPK pathway by increasing the expression of p38, JNK and ERK as well as the expression of p-p38, p-JNK and p-ERK in testis and TM3 cells. The p38 molecular inhibitor (SB203580) significantly alleviated the T-2 toxin-induced decrease in testosterone synthase expression in TM3 cells and the T-2 toxin-induced reduction in testosterone content in TM3 cell culture supernatants. In summary, p38 mediates T-2 toxin-induced Leydig cell testosterone synthesis disorder.

P-002 Zinc restores testicular integrity and function in HAART-treated male Wistar rats via modulation of Nrf2/NFkB pathway and downregulation of caspase 3 signaling

Abstract Study question Will zinc treatment attenuate highly active antiretroviral drugs (HAART)-induced testicular dysfunction when used concomitantly? Summary answer Zinc blunted HAART-induced reproductive toxicity by attenuating testicular and epididymal oxido-inflammatory injury via modulation of Nrf2/NFkB signaling and downregulation of caspase 3-mediated apoptosis. What is known already The use of HAART has been established to be effective in the management of HIV; however, it has also been reported to induce male reproductive dysfunction. Studies have revealed that HAART impairs fertility by reducing libido and inducing testicular and sperm damage via an oxidative stress-sensitive pathway. On the other hand, zinc has been shown to enhance reproductive function by improving testicular redox state. Till date, no study has reported the impact of zinc co-administration with HAART on testicular integrity and function, spermatogenesis and sperm quality, and sexual behaviour. Study design, size, duration This is a prospective experimental study using animal model. Forty sexually mature inbred male Wistar rats of comparable age and weight were used for the study. The study lasted 8 weeks. Participants/materials, setting, methods Animals were acclimatized for two weeks, then randomly assigned into four groups (n = 10). The control rats received 0.5mL of distilled water as vehicle, zinc-treated animals received zinc supplement, HAART-treated rats received a cock-tail of antiretroviral drugs (Efavirenz, Lamivudine, and Tenofovir), while the Zn + HAART-treated rats received treatment as zinc-treated as well as HAART-treated. The doses of drugs used were the Human Equivalent doses for rats. Main results and the role of chance Zinc blunted HAART-induced rise in testicular activities of gamma glutamyl transferase, lactate dehydrogenase activity, concentrations of lactate, and reduced testicular sorbitol dehydrogenase activity. Also, zinc ameliorated HAART-induced decline in the activities of testicular and epididymal superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase, as well as glutathione and Nrf2 concentrations, and rescued HAART-induced increase in testicular and epididymal concentrations of uric acid, malondialdehyde, 8-OHdG, xanthine oxidase and MPO activities, and TNF-α, IL-1β, and NF-kB levels. This was associated with dampening of HAART-led upregulation of caspase 3 activity by zinc. Furthermore, zinc alleviated HAART-led reduction in spermatogenesis and sperm quality, serum nitric oxide and penile cGMP, and circulatory levels of FSH, LH, and testosterone, as well as testicular concentrations of 3β-HSD, 17β-HSD, and testosterone. In addition, zinc prevented HAART-induced sexual dysfunction and poor fertility indices. Limitations, reasons for caution This study was conducted in a rat model; hence findings should be extrapolated to human with care. Thus, clinical trials are recommended to validate these findings. Wider implications of the findings For the first time, we demonstrated the protective role of zinc treatment on HAART-induced male reproductive dysfunction. These findings provide further mechanisms through which HAART induces male infertility, and the possible protective role of zinc. Trial registration number not applicable

Blepharis persica increases testosterone biosynthesis by modulating StAR and 3β-HSD expression in rat testicular tissues

Objective: To evaluate the effect of methanolic extract and ethyl acetate fraction of methanol extract prepared from the seeds of Blepharis (B.) persica on testosterone biosynthesis and also to elucidate the underlying mechanism. Methods: Forty-eight male Wistar rats were divided into eight groups (n=6 per group). Group I received 0.3% w/w gum acacia suspension p.o. and served as the normal control group. Group II was administered testosterone propionate in arachis oil i.m. as the positive control group. Group III to V received B. persica methanolic extract p.o. at doses of 50, 100 and 200 mg/kg body weight. Group VI to VIII received B. persica ethyl acetate fraction p.o. at doses of 50, 100 and 200 mg/kg body weight. The testis was used for biochemical estimation and histological studies. The effects of methanolic extract and ethyl acetate fraction of B. persica on testicular testosterone, mRNA expression corresponding to steroidogenic acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) along with 3β-HSD enzyme assay were evaluated in testicular tissues and sperm concentration. Ethyl acetate fraction of B. persica was subjected to column chromatography. In-vitro studies were performed using TM3 cell line at three dose levels (50, 100, 200 μg/mL), each for methanolic extract, ethyl acetate fraction and 2-benzoxazolinone for evaluation of their comparative effect on testosterone production. Results: Ethyl acetate fraction and methanolic extract of B. persica could elevate the testicular testosterone content compared to the normal control group. The treatment with methanolic extract and ethyl acetate fraction of B. persica increased the expression of mRNA corresponding to StAR by 6.7 fold and 10.6 fold, respectively, whereas the mRNA expression of 3β-HSD increased by 5.7 fold and 7.3 fold, respectively. Moreover, fraction and extract treatment exhibited increased 3β-HSD activity in the testicular tissues and were found to elevate sperm concentration in seminal fluid. The spermatogenic potential was further ensured by histological observations. 2-benzoxazolinone was isolated from ethyl acetate fraction and identified using spectral studies. It showed the ability to increase the testosterone content in the TM3 Leydig cells. Conclusions: Methanolic extract and ethyl acetate fraction of B. persica are able to increase the testicular testosterone in rats by elevating mRNA expression of StAR and 3β-HSD in testicular tissues, leading to increase the sperm concentration.

Evidence of melatonin ameliorative effects on the blood-testis barrier and sperm quality alterations induced by cadmium in the rat testis

Herein, we further document the protective action of melatonin (MLT) in mitigating cadmium (Cd) effects on adult rat testis. Cd treatment provoked testicular injury, that was documented by histological and biomolecular alterations, i.e., decrease of serum and testicular testosterone concentration and modified sperm parameters. Mainly, both the cytoarchitecture of the blood-testis barrier (BTB) and germ cell morphology were perturbed, as highlighted by impairment in structural (OCN, VANGL, Cx43) and regulative (Src and FAK) protein levels and/or activation. The study focused on the involvement of the autophagy pathway, that was enhanced especially in the Sertoli cells, probably in response to the disorganization of the BTB. Results obtained with the MLT co-treatment demonstrated that its administration decreased the level of oxidative damage caused by Cd, with reversal of all the observed changes. Moreover, the beneficial effects of MLT alone were evidenced by an increase of sperm quality, in term of motility and DNA integrity. The combined results, obtained in rat, strongly encourage to consider a role for MLT in improving also human testicular health, not only in men exposed to Cd, but also in those having fertility disorders, to ameliorate sperm quality and, consequently, reproductive success.

Sexual behaviour, semen quality and fertility of young Border Leicester rams administered melatonin during spring

This study was conducted to test the hypothesis that between year variability in reproductive response to melatonin treatment of young Border Leicester (BL) rams in spring was related to sexual maturity at the time of treatment. Two variables of sexual maturity (mating behaviour and semen quality) were examined. In Experiment 1, mating activity of melatonin-treated and untreated rams was examined at two sites before a flock mating at one site. In Experiment 2, testosterone concentrations, semen quality and variables of in vitro fertilising capacity were examined. In Experiment 1, melatonin treatment did not alter sexual behaviour (latency to nose, total nosings, courtings including Flehmen expression, attempted mounts, mounts, ejaculations) with the exception of a lesser (P < 0.05) Flehmen expression at Site 1 whereas values for most variables were greater (P < 0.05) at Site 2. Treatment resulted in a greater pregnancy rate (89 % and 82 %, respectively; P < 0.05) and advanced distribution of pregnancies (P < 0.001) at Site 1. Testicular size and testosterone concentrations were also greater (P < 0.05) as a result of melatonin treatment in spring whilst the reverse occurred in autumn. In Experiment 2, sperm motility and in vitro fertilising capacity were greater (P < 0.05) and acrosome damage less (P < 0.05) as a result of melatonin treatment. In conclusion, variability in age at sexual maturity of young BL rams at spring mating can be reduced with melatonin treatment and is associated with differences in sexual behaviour, testicular growth, testosterone concentrations, and sperm quality.

Clinical evaluation of chemo-sterilization through histomorphology and hormonal changes in bucks

The study was aimed to evaluate methods for nonsurgical castration of Black Bengal bucks by intra-testicular injection of calcium chloride, sodium chloride, citric acid solutions or sterile deionized water. Twelve healthy bucks were randomly allotted to groups A, B, C, and D, consisting of three bucks each. The local anesthetic, 2% lidocaine hydrochloride, was infused into the spermatic cord of each buck, followed by bilateral intra-testicular injections of 30% calcium chloride (CaCl2), 25% sodium chloride (NaCl), 50% citric acid (C6H8O7) solutions, and sterile deionized water dosed at 2 ml per testis in groups A, B, C and D respectively. To evaluate the efficacy of chemical agents on the inactivation of testes, clinical parameters, changes in scrotal circumference, testicular fine needle aspiration (TFNA), histopathology and serum concentration of testosterone and LH were monitored. A significant decrease in the scrotal circumference was observed between the intra-testicular injection and day 14 in all the bucks. Absence of spermatogenic cells and spermatozoa in the testicular biopsy was observed on day 14 post injection in the bucks, except for one in group C. Histopathology revealed massive destruction of seminiferous tubules and disorganization of the testicular parenchyma. Serum testosterone concentration declined significantly on day 14 compared with day 0. Consequently, the gradual elevation in serum LH concentration was significant. Thus, intra-testicular injections of CaCl2 and NaCl were more effective than C6H8O7 in inducing chemical-based nonsurgical castration.

Protective Effects of Hu-Lu-Ba-Wan () against Oxidative Stress in Testis of Diabetic Rats through PKCα/NAPDH Oxidase Signaling Pathway.

To explore the protective effect and the underlying mechanism of Hu-Lu-Ba-Wan (, HLBW) on the testis of diabetic rats. Twenty-four male Wistar rats (160-180 g) were randomly divided into 3 groups according to a random number table, including a control group (n=8), diabetic group (n=8), and HLBW group (n=8). Diabetic rat model was established by high-fat-diet administration and single intravenous injection of streptozotocin (26 mg/kg). Then HLBW granule was administrated for 12 weeks. Fasting blood glucose and insulin levels as well as serum total testosterone level and testicular testosterone content were examined. Oxidative stress markers in both serum and testis were tested. Meanwhile, testicular morphology was observed under hematoxylin and eosin (HE) and the ultrastructure of Leydig cell was observed by electron microscope. The superoxide anion level was detected by DHE, and TUNEL-positive cells of testis was evaluated by TUNEL assay. The gene and protein expression of protein kinase C (PKCα), phosphorylated PKCα (P-PKCα) and P47phox in testicular tissues were determined by quantitative RT-PCR analysis and Western bolt analysis. Compared with the diabetic group, HLBW treatment significantly reduced the fasting glucose levels and increased the levels of fasting insulin and testosterone in serum (P<0.01). HLBW administration also reduced the levels of reactive oxygen species (ROS) in plasma and alleviated the damage of oxidative stress in the testis of diabetic rats. Additionally, HLBW down-regulated the protein and mRNA levels of PKCα, P-PKCα and P47phox in testicular tissues. HLBW may attenuate the oxidative stress in the testis of diabetic rats via PKCα /NAPDH oxidase signaling pathway.

Testicular Testosterone and Estradiol Concentrations and Aromatase Expression in Men with Nonobstructive Azoospermia.

Spermatogenesis is strictly regulated by the intratesticular hormonal milieu, in which testosterone (T) and estradiol (E2) play pivotal roles. However, the optimal expression of aromatase and intratesticular T (ITT) and E2 (ITE2) levels are unknown. To investigate ITT/ITE2 and aromatase expression in men with nonobstructive azoospermia (NOA) and to elucidate the roles of aromatase in spermatogenesis, as determined based on sperm retrieval by microdissection testicular sperm extraction (micro-TESE). A retrospective study at a reproductive center using serum, testicular specimens, and intratesticular fluid. Seventy-six men with NOA, including 4 men who received 3 months of anastrozole administration prior to micro-TESE, and 18 men with obstructive azoospermia. Testicular aromatase expression was evaluated using immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (RT-PCR). Intratesticular T and ITE2 levels were determined using liquid chromatography-tandem mass spectrometry. Aromatase was mainly located in Leydig cells, and the levels of its transcript and protein expression levels were increased in men with NOA. No correlation was observed between serum T/E2 and ITT/ITE2 levels, whereas significant associations were observed between decreased ITT and increased ITE2, aromatase expression, and sperm retrieval. Treatment with anastrozole increased the ITT/ITE2 ratio and decreased aromatase expression. A close association between the expression of aromatase in Leydig cells and ITT/ITE2 was shown. Leydig cell aromatase is a factor that is independently correlated with spermatogenesis, and aromatase inhibitors may open a therapeutic window by increasing ITT/ITE2 in selected patients.

Dietary Exposure to Oxidized Frying Oil from Fetus to Adulthood Suppresses Male Reproductive Development by Altering Testicular Cholesterol and Testosterone Homeostasis in Sprague Dawley Rats

Dietary frying oil may have endocrine-disrupting effects, as a feminization effect was observed in cohorts of C57BL/6J male mice fetuses from dams consuming oxidized frying oil (OFO) during pregnancy. The aim of present study was to test the hypothesis that OFO is an anti-androgen. In experiment 1, male progeny of Sprague Dawley female rats fed fresh oil or an OFO diet (10g fat/100g, from fresh or 24-h-fried soybean oil; [control diet (C) and OFO groups, respectively] from midgestation through lactation were studied. Pups were weaned at 3 wk of age and then consumed their mothers' diet until 9 wk of age. In addition, a group of dams and pups that consumed a high-fat diet (HF; 10g fried and 20g fresh soybean oil/100g) was included to counteract body-weight loss associated with OFO ingestion. Indices of male reproductive development and testosterone homeostasis were measured. In experiment 2, male rats were allocated to C and OFO groups (treated as above) and indices of male fertility compared at 9-10 wk of age. In experiment 1, final body weights of the HF group were lower (17%) than the C group but higher (14%) than the OFO group (P< 0.0001 for each). In addition to abnormalities in seminiferous tubules, HF and OFO groups did not differ from one another, but, compared with the C group, haddelayed preputial separation (4.9 d)andreductions inserum testosterone concentrations (17-74%), anogenital distance (8-20%),weights of androgen-dependent tissues (8-30%), testicular testosterone and cholesterol concentrations (30-40%), and mRNA levels of genes involved in steroidogenesis and cholesterol homeostasis (30-70%). In experiment 2, OFO-exposed males had 20% lower sperm motility (P< 0.05); however, when mated to normal females, pregnancy rates and litter sizes did not differ between OFO and C groups. The anti-androgenic effect of OFO in Sprague Dawley rats was attributed to decreased testicular concentrations of cholesterol (testosterone precursor) and notbody-weight loss.

Low Dose of Genistein Alleviates Mono-(2-Ethylhexyl) Phthalate-Induced Fetal Testis Disorder Based on Organ Culture Model.

Mono-(2-ethylhexyl) phthalate (MEHP) and genistein have been classified as endocrine-disrupting chemicals (EDCs) which interfere with the differentiation and development of the male reproductive system. However, how these two EDCs would affect fetal rat testis development at a low dose was rarely studied. In this study, we established the organ culture system and applied it to evaluate testicular effects following multiple EDC exposure at a low dose. 15.5 days postcoitum fetal rat testes were dissected, cultured, and exposed to vehicle (control), GEN (1 μmol/L, G), MEHP (1 μmol/L, M), or GEN (1 μmol/L)+MEHP (1 μmol/L, G+M). Testicular cell markers, testosterone concentration, redox state, testicular histology, and testicular ultrastructure were evaluated. Our results showed that a low dose of MEHP suppressed the development of Sertoli cells, Leydig cells, and gonocytes by triggering oxidative injuries, which was consistent with the ultrastructural findings. However, coadministration of genistein at a low dose could partially attenuate MEHP-induced fetal testis damage through antioxidative action. Cotreatment of genistein at a low dose may have a promising future on its protecting role for attenuating other EDC-induced reproductive disorders during early life. Based on the results, it can be speculated that dietary intake of isoflavones may make the fetal testis less susceptible to phthalate-induced injury.

Icariin Improves Age-Related Testicular Dysfunction by Alleviating Sertoli Cell Injury via Upregulation of the ERα/Nrf2-Signaling Pathway.

Sertoli cells play crucial roles in spermatogenesis and are impaired by aging. Icariin, a flavonoid from Epimedium, has been reported to exhibit anti-aging effects and improve testicular dysfunction in the clinical setting. However, whether icariin improves age-related degeneration of testicular function via protection from Sertoli cell injury remains unclear. In the present study, we evaluated the protective effect of icariin on Sertoli cell injury and explored the possible mechanism(s) in vivo and in vitro. Dietary administration of icariin for 4 months significantly ameliorated the age-related decline in testicular function by increasing testicular and epididymal weights and indices, sperm count and sperm viability, testicular testosterone and estradiol concentrations, and seminiferous tubule diameters and heights. In addition, icariin protected age-related Sertoli cells from injury as evidenced by an analysis of Sertoli cell number, ultrastructure, and function. Such changes were accompanied by upregulation of ERα and Nrf2 signaling in Sertoli cells. Parallel in vitro studies also demonstrated that icariin inhibited untoward effects on the TM4 mouse Sertoli cell line with concomitant upregulation of ERα and Nrf2 signaling. Conversely, ERα siRNA reversed icariin-mediated protection of Sertoli cell injury. Our data suggest that icariin effectively ameliorates age-related degeneration of testicular function by alleviating Sertoli cell injury via the ERα/Nrf2 signal-transduction pathway. Thus, mitigating Sertoli cell damage via the ERα/Nrf2 signaling pathway likely represents a promising strategy for the prevention of age-related testicular dysfunction.

Testicular hyperthermia reduces testosterone concentrations and alters gene expression in testes of Nelore bulls

Increased testicular temperature reduces sperm motility, morphology and fertility. Our objectives were to characterize effects of testicular hyperthermia (scrotal insulation) on acute testosterone concentrations and gene expression in Bos indicus testes. Nelore bulls (n = 20), ∼27 mo of age, 375 kg, scrotal circumference >31 cm, with ≥30% motile sperm, were allocated into four groups (n = 5/group): non-insulated (Control) and insulation removed after 12, 24, or 48 h. Immediately after insulation, intratesticular temperatures (needle thermocouples) were coolest in Control bulls and warmest in 48-h bulls (mean ± SEM, 35.28 ± 0.31 vs 38.62 ± 0.57 °C, P < 0.05). Bulls were castrated and testes recovered. Testicular testosterone concentrations were higher in Control versus 48-h bulls (3119 ± 973.3 and 295.5 ± 122.8 ng/g of tissue, respectively, P < 0.05). Total RNA was extracted, reverse transcribed and RT-qPCR done. For STAR, mRNA abundance decreased from Control to 48 h (1.14 + 0.32 vs 0.32 + 0.5, P < 0.05). For BCL2, expression decreased from Control to 24 h (1.00 + 0.07 vs 0.70 + 0.12, P < 0.05), but then rebounded. In addition, GPX1 had a 70% increase (P < 0.05) at 48 h, whereas HSP70 had a 34-fold increase (P < 0.05) at 12 h and 2- and 14-fold increases (P < 0.05) at 24 and 48 h, respectively. HSF1, BAX, P53 and CASP 8 remained unchanged. Downregulation of STAR, critical in androgen production, was consistent with reduced testosterone concentrations, whereas increased GPX1 enhanced testicular antioxidative capability. Huge increases in HSP70 conferred protection again apoptosis and cell destruction, whereas reduced BCL2 promoted apoptosis. These findings provided novel insights into acute tissue responses (testosterone and gene activity) to testicular hyperthermia in B. indicus bulls.

Influence of Different Feeding Ratio of Pomegranate Peels Powder on Testicular Dimensions, Body Weight and Testosterone Concentration in Karadi Male Lambs

The current study was presented to conclude the influence of different ratio of pomegranate peel powder (PP) in feed of Karadi male lambs, they divided to 4 groups (0% as a control, 1% treatment 1, 2% treatment 2 and 4% as treatment 3), on scrotal circumference (SC), scrotum height (SH), body weight (BW) and blood serum testosterone concentration (T). In this study, sixteen (16) of Karadi male lambs used with 24.30 ± 0.40 kg weight and with 4-5 months old, all lambs have been randomly divided equally into four (4) groups and individually penned in a box for during 63 days. Results in our study showed that SC was significantly different and recorded higher 17.875±0.54 cm in group that fed on 1% PP, also SH recorded higher and significant different in group fed on 1% PP compared with other groups 10.000±0.32 cm. Average blood serum testosterone reduces in all groups compared to control group that fed on 0% of PP (11.418±0.15 n.mol/L). Final body weight in our study had significantly different (P &lt; 0.05) between groups, 1% PP recorded higher weight (34.9 ± 0.58) kg. Each average daily gain, total body gain and initial body weight had not significant different in all groups. Correlation coefficient between SC, SH, blood serum testosterone concentration and final BW computed and ranged between (-0.397 to + 0.914).

Effect of bisphenol F, an analog of bisphenol A, on the reproductive functions of male rats

ObjectiveBisphenol A (BPA) is a monomer primarily used in the production of polycarbonate plastic and epoxy resins. Bisphenol F (BPF) is apparently the main BPA replacement that is used increasingly. BPF has been detected in canned food, thermal paper receipts, and soft drinks. In the present experiment, we did both in vitro and in vivo studies to evaluate the effect of low and high-dose BPF exposures on testosterone concentration, oxidative stress, and antioxidants activity in reproductive tissues of male rats.MethodsAdult (80–90 days old) male Sprague Dawley rats (n = 36) obtained from the rodent colony of Animal Sciences Department of Quaid-i-Azam University. The direct effects of BPF on the antioxidant enzymes and testosterone secretion were measured in vitro and in vivo studies. In an in vivo experiment, adult male Sprague Dawley rats (n = 42) were exposed to different concentrations of bisphenol F (1, 5, 25, and 50 mg/kg/d) for 28 days. Various biochemical parameters were analyzed including the level of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), reactive oxygen species (ROS), and lipid peroxidation (LPO). Moreover, sperm motility, daily sperm production (DSP), comet assay, and histological analysis were performed.ResultsIn vitro study showed that BPF exposure significantly (p < 0.05) induced oxidative stress biomarkers, i.e., ROS and LPO, while it did not change antioxidant enzyme and testicular testosterone concentration. Whereas, an in vivo study revealed that BPF induced dose-dependent effect and high-dose (100 mg/kg) exposure of BPF significantly reduced tissue protein (p < 0.05) content, CAT (p < 0.001), SOD (p < 0.05), and POD (p < 0.05) levels while significantly (p < 0.05) augmented ROS and lipid peroxidation. Furthermore, BPF reduces testosterone, LH, and FSH secretion in a dose-dependent manner. Significant (p < 0.001) reduction in plasma and intra-testicular testosterone, LH, and FSH was noticed at 100 mg/kg BFP dose. High-dose exposure reduces spermatogenesis.ConclusionBPF showed an antagonistic effect on male reproductive hormones and induce alterations in testicular morphology. Increased oxidative stress and decreased testicular antioxidant status might be the underlying mechanism of BFP-induced testicular toxicity.