Reduced Serum Testosterone Research Articles

Semen quality in hypogonadal acromegalic patients.

Growth hormone (GH) activity might be implicated in male reproductive function. One previous study has suggested significantly reduced semen quality in untreated acromegalic patients due to both reduced sperm counts and sperm motility. A retrospective study comprising ten uncontrolled hypogonadal acromegalic patients (median age 29years) who delivered semen for cryopreservation before initiation of testosterone therapy. Semen variables and hormone concentrations were compared to those of ten non-acromegalic hypogonadal men with pituitary disease (age 31years) and those of young healthy men. Acromegalic patients vs. non-acromegalic patients had a higher percentage of progressive motile spermatozoa (62 vs. 47%, p = 0.04). Eight of ten acromegalic patients and 82% of controls had total sperm counts above 39 million and progressive motile spermatozoa above 32% (p = 0.55), corresponding to the WHO 2010 reference levels for expected normal fertility for these variables. Non-acromegalic patients vs. healthy controls had reduced percentage of progressive motile spermatozoa (47 vs. 57%, p = 0.02) and only five of ten patients had semen quality above the WHO reference level, which was significantly lower than observed in healthy controls (p = 0.022). Total sperm counts were similar between both patient groups and controls. There were no differences in reproductive hormone levels between acromegalic patients vs. non-acromegalic patients (p-values between 0.10 and 0.61). Compared to healthy controls both patient groups had severely reduced serum testosterone, calculated free testosterone. Despite severe hypoandrogenism acromegalic patients had semen quality similar to healthy controls based on determination of the number of progressively motile spermatozoa. By contrast non-acromegalic pituitary patients had reduced sperm motility. Our data do not support reduced semen quality in acromegaly.

Marjoram and sage oils protect against testicular apoptosis, suppressed Ki‐67 expression and cell cycle arrest as a therapy for male infertility in the obese rats

Alterations in testicular apoptosis, cell cycle progression and proliferation rate in dietary-induced obese male rats and role of oral administration of marjoram (0.16ml/kg BW) and sage (0.05ml/kg BW) oils were evaluated. Results showed increased body weight, serum leptin, testicular lipid peroxidation, protein oxidation and nitric oxide, with reduction in serum testosterone, sperm count and endogenous enzymatic and non-enzymatic antioxidants in testis of the obese rats. Flow cytometry results revealed increased number of annexin-V (+ve) cells with activation of apoptotic proteins (Bax, caspase-3) and reduction of anti-apoptotic Bcl-2. Cell cycle arrest at phases S and G2/M with decline in expression of Bcl-2 and germ cell proliferation marker ki-67 was also validated, indicating lowered spermatogenesis in the obese rats. Supplementation of marjoram or sage oils displayed normalized body weight, sperm count, germ cells apoptosis and proliferation, suggesting the two oils as a new therapeutic approach against obesity promoted male infertility. PRACTICAL APPLICATIONS: Obese men have a greater chance of fertility problems compared to those with normal weight. Obesity-associated oxidative stress and free radicals production have shown to adversely affect sperm quality with activation of pro-apoptotic pathways, allowing germ cell death. Marjoram and sage essential oils are now being widely studied due to their antioxidant and radical scavenging properties. Our findings indicated effectiveness of the two oils for combating body weight gain, testicular oxidative stress, and apoptosis, which seemed to aid in increasing sperm count. The outcomes of this study may help scientists to formulate novel medications for improving fertility problems in men.

Dimethoate blocks pubertal differentiation of Leydig cells in rats.

Dimethoate is an organophosphate pesticide. It is widely used in agriculture. However, whether it blocks pubertal development of Leydig cells remains unknown. In the current study, we exposed male Sprague Dawley rats with 7.5 and 15 mg kg-1 dimethoate from postnatal day 35-56. We also exposed Leydig cells isolated from 35-day-old rats for 3 h. Dimethoate reduced serum testosterone levels at 7.5 and 15 mg kg-1 but increased serum luteinizing hormone and follicle stimulating hormone levels at 15 mg kg-1. Dimethoate did not influence Leydig cell number but reduced Leydig cell size and down-regulated Star, Cyp11a1, and Hsd3b1 in Leydig cells as well as their protein expression. Dimethoate inhibited basal androgen output in a dose-dependent manner with the inhibition starting at 0.05 μM. It significantly inhibited luteinizing hormone and 8Br-cAMP stimulated androgen outputs at 50 μM. It significantly inhibited 22R-hydroxycholesterol and progesterone-mediated androgen outputs at 50 μM. Further study demonstrated that dimethoate also down-regulated the expression of Star, Cyp11a1, and Hsd3b1 at 5 or 50 μM invitro. Dimethoate did not directly inhibit rat testicular steroidogenic enzyme activities at 50 μM. In conclusion, dimethoate targets Star, Cyp11a1, and Hsd3b1 transcription, thus blocking Leydig cell differentiation during puberty.

Aflatoxin B1 promotes autophagy associated with oxidative stress-related PI3K/AKT/mTOR signaling pathway in mice testis

Aflatoxin B1 (AFB1) is a hazard environmental pollutants and the most toxic one of all the aflatoxins. AFB1 can cause a serious impairment to testicular development and spermatogenesis, yet the underlying mechanisms remain inconclusive. Oxidative stress acts as a master mechanism of AFB1 toxicity, and can promote autophagy. Abnormal autophagy resulted in testicular damage and spermatogenesis disorders. The objective of this study was to explore the effect of AFB1 on autophagy in mice testis and its potential mechanisms. In this study, male mice were intragastrically administered with 0, 0.375, 0.75 or 1.5 mg/kg body weight AFB1 for 30 days. We found that AFB1 induced testicular damage, reduced serum testosterone level and impaired sperm quality accompanied with the elevation of oxidative stress and germ cell apoptosis. Interestingly, we observed increasing numbers of autophagosomes in AFB1-exposed mice testis. Meanwhile, AFB1 caused testis abnormal autophagy with the characterization of increased expressions of LC3, Beclin-1, Atg5 and p62. Furthermore, AFB1 downregulated the expressions of PI3K, p-AKT and p-mTOR in mice testis. Taken together, our data indicated AFB1 induced testicular damage and promoted autophagy, which were associated with oxidative stress-related PI3K/AKT/mTOR signaling pathway in mice testis.

Trimethyltin (TMT) Reduces Testosterone Production in Adult Leydig Cells in Rats.

Trimethyltin (TMT) is widely used as a plastic heat stabilizer and can cause severe toxicity. Here, the effects of TMT on testosterone production by adult Leydig cells and the related mechanisms of action were investigated. Eighteen adult male Sprague Dawley rats (56 days old) were randomly divided into 3 groups and given intraperitoneal injection of TMT for 21 consecutive days at the doses of 0 (vehicle control), 5, or 10 mg/kg/d. After treatment, trunk blood was collected for hormonal analysis. In addition, related gene and protein expression in testes was detected. At 10 mg/kg, TMT significantly reduced serum testosterone levels but increased serum luteinizing and follicle-stimulating hormone levels. The messenger RNA and protein levels of luteinizing hormone/chorionic gonadotropin receptor, steroidogenic acute regulatory protein, cytochrome P450 17-hydroxylase/17,20-lyase, follicle-stimulating hormone receptor, and SRY box 9 were significantly lower in the TMT-treated testes than in controls. Immunohistochemical study showed that TMT decreased adult Leydig cell number. In conclusion, these findings indicate that TMT reduced adult Leydig cell testosterone production in vivo by directly downregulating the expression of steroidogenic enzymes and decreasing adult Leydig cell number in the testis.

Vitamin B12 Prevents Cimetidine-Induced Androgenic Failure and Damage to Sperm Quality in Rats.

Cimetidine, used as an anti-ulcer and adjuvant treatment in cancer therapy, causes disorders in the male reproductive tract, including steroidogenesis. However, its effect on sperm quality and male fertility has been poorly addressed. Since vitamin B12 has demonstrated to recover spermatogonia number and sperm concentration in cimetidine-treated rats, we evaluated the impact of cimetidine on sperm quality and fertility potential and whether vitamin B12 is able to prevent the harmful effect of this drug on steroidogenesis and sperm parameters. Adult male rats were treated for 52 consecutive days as follows: cimetidine group (100 mg/kg of cimetidine), cimetidine/vitamin B12 group (100 mg/kg of cimetidine + 3 μg vitamin B12), vitamin B12 group (3 μg vitamin B12) and control group (saline). Serum testosterone levels and immunofluorescence associated to western blot for detection of 17β-HSD6 were performed. Sperm morphology and motility, mitochondrial activity, acrosome integrity, DNA integrity by Comet assay, lipid peroxidation as well as fertility potential were analyzed in all groups. Apoptotic spermatids were also evaluated by caspase-3 immunohistochemistry. In the cimetidine-treated animals, reduced serum testosterone levels, weak 17β-HSD6 levels and impaired spermiogenesis were observed. Low sperm motility and mitochondrial activity were associated with high percentage of sperm tail abnormalities, and the percentage of spermatozoa with damaged acrosome and DNA fragmentation increased. MDA levels were normal in all groups, indicating that the cimetidine-induced changes are associated to androgenic failure. In conclusion, despite the fertility potential of rats was unaffected by the treatment, the sperm quality was significantly impaired. Therefore, considering a possible sperm-mediated transgenerational inheritance, the long term offspring health needs to be investigated. The administration of vitamin B12 to male rats prevents the androgenic failure and counteracts the damage inflicted by cimetidine upon sperm quality, indicating that this vitamin may be used as a therapeutic agent to maintain the androgenic status and the sperm quality in patients exposed to androgen disrupters.

Clinical studies investigating the use of leuprorelin for prostate cancer in Asia.

BackgroundLeuprorelin is a well-established treatment for prostate cancer (PCa); however, there is limited information on its use in Asian males. This review of English language publications between January 2000 and 2016 describes the outcomes of clinical trials on leuprorelin in Asian males with PCa of any grade, stage, or histopathology.MethodsThe literature search was undertaken using PubMed, Cochrane Library, and ClinicalTrials.gov databases.ResultsWe identified nine studies from Japan, two studies from South Korea, and one international, multisite study which included Asian sites, with a total of 1,652 males previously diagnosed with PCa. All studies included subcutaneous or depot administration of leuprorelin at varying dose levels including 3.75 mg four weekly, 11.25 mg 12 weekly, or 22.5 mg every 12 or 24 weeks. Leuprorelin was administered as monotherapy or in combination with chemotherapy or hormonal therapy. Leuprorelin appears well tolerated in Asian males and is effective in reducing serum testosterone to castration levels (<50 ng/dL (<1.7 nmol/L)) and prostate-specific antigen levels. Common adverse events included hot flushes and mild hepatic dysfunction. Leuprorelin was shown to provide reasonable survival rates in PCa (T1b-T3N0M0) and in metastatic disease; another reasonable option for these patients is radiation therapy. Leuprorelin treatment also improved the quality of life.ConclusionLeuprorelin may be an appropriate and efficacious treatment for males with PCa (T1b-T3N0M0). Leuprorelin treatment was well tolerated and associated with improvement in the quality of life.

AMEOLARATIVE EFFECT OF ALOE VERA GEL ON TRAMADOL REPRODUCTIVE TOXICITY IN ADULT ALBINO RAT Rabab Fawzy Hindawy1, Nagah Elsayed Mohammed Ali 1and Fatma Fawzi Hendawy2 1Forensic Medicine &amp; Clinical Toxicology Department, and2 Pharmacology Department, Faculty of Medicine, Benha University, Egypt.

ABSTRACT Background: Tramadol is an opioid pain medication used to treat premature ejaculation and as an antidepressant also young population abuse tramadol. Long-term administration of tramadol has negative impacts on many organs like ovary and testis. Objective: The aim of study was to assess effect of tramadol on testes and ovaries of adult albino rats and to elucidate possible protective effect of Aloe Vera gel. Methodology: one hundred adult albino rats were used in present study. Animals were divided into five groups: control group, solvent control group, Aloe Vera gel group, tramadol group and tramadol and Aloe Vera gel group, testes and ovaries were excised and specimens were processed for light, electron microscopic examination, blood samples were obtained to be tested for: testosterone, malondialdehyde MDA, and reduced glutathione GSH. Results: The present work detected a highly significant reduction in serum testosterone, and GSH levels and a highly significant increase in serum MDA levels in tramadol group as compared to control group. A highly significant increase in serum testosterone, and GSH levels and a highly significant reduction in MDA levels in tramadol and Aloe Vera gel group as compared to group tramadol and these results were non-significant as compared to controls. Tramadol-treated male rats showed dilatation in seminiferous tubules and vacuolation, testicular atrophy, and spermatogenic arrest, ovaries of tramadol-treated rats showed atretic follicles, with hyperplasia of corpus luteum (CL) and vacuolation. All histopathological changes, were ameliorated when Aloe Vera was co-administered with tramadol. Conclusion: Tramadol has harmful effect on testes and ovaries of rats. Administration of Aloe Vera accelerates healing of testicular tissue structure and has positive effect on testosterone level.

Combined effects of arsenic exposure and diabetes on male reproductive functions.

It is known that exposure to either arsenic or hyperglycemia can induce male reproductive damages. However, their combined effects on male reproductive organs are still unclear. Therefore, the present study investigated morphological and functional parameters of the testis, epididymis, and spermatozoa in diabetic rats exposed to arsenate. Diabetes was induced in male rats by intraperitoneal streptozotocin injection. While a set of healthy and diabetic animals received saline solution (negative control and diabetes control, respectively), the other set received 10mg/L sodium arsenate (arsenic control and diabetes + arsenic groups, respectively) for 40days in drinking water. Testosterone concentration, daily sperm production, sperm counts in the testis and epididymis, and sperm parameters were evaluated in the groups. Moreover, testis and epididymis were subjected to antioxidant enzymes analysis, micromineral determination, and histopathological evaluation. Arsenate exposure reduced serum testosterone concentration in healthy animals and worsened this reduction in diabetic rats. In addition, the number of spermatozoa in testis and epididymis tissues, as well as the daily sperm production, was decreased in these groups. Sperm parameters such as motility, morphology, and integrity of acrosomal and plasma membranes were impaired in health animals exposed to arsenate. The combination of diabetes and arsenate, in turn, increased only the percentage of spermatozoa with abnormal morphology. Moreover, the proportion of arsenic increased in the testis and epididymis of both groups receiving arsenate. Its bioaccumulation in these organs caused an imbalance in antioxidant enzymes activities and mineral content in healthy animals, enhancing these changes in diabetic rats. Testicular pathologies occurred mainly in animals co-exposed to diabetes and arsenate. Our results indicate that arsenate exposure enhances several damages to male reproductive functions in diabetic rats, mainly by impairing testosterone levels and inducing nitrosative stress in testis and epididymis.

Maternal exposure to bisphenol A during pregnancy interferes testis development of F1 male mice.

This study was conducted to investigate the effects of maternal exposure to bisphenol A (BPA) on testis development of F1 male mice. The BPA exposure model of pregnant mice was prepared by intragastric administration of BPA at the doses of 0, 2.5, 5, 10, 20, and 40mg/kg/day at gestation day (GD) 0.5-17.5. The testis index of the offspring mice was calculated at postnatal day (PND) 21 and PND 56. The results showed that maternal exposure to 20mg/kg BPA during pregnancy significantly increased the testicular index of F1 males at PND 21, and 40mg/kg BPA significantly decreased the testicular index of F1 males at PND 56 (P < 0.01). BPA significantly reduced serum testosterone (T) and estradiol (E2) levels, and improved testicular ERα and ERβ levels in F1 males at both PND 21 and PND 56. BPA exposure also upregulated transcription of testicular Dnmt1 and inhibited the transcription of testicular Dnmt3A and Dnmt3B in F1 mice at PND 21. BPA reduced the transcriptional level of testicular DNA methyltransferase (Dnmt), increased the expression of testicular caspase-7, caspase-9, and bax, and decreased the expression of bcl-2 in F1 mice at PND 56. Consistent with that, BPA improved the apoptosis rate in the testis at PND 56 (P < 0.01 or P < 0.05). Our study indicates that BPA disrupts the secretion of testosterone, estradiol, and estrogen receptors by interfering with the transcription of testicular DNA methyltransferase (Dnmt) in offspring males, which damages testicular tissues and affects the potential reproductive function.

SAT-353 The Synthetic Vitamin D Analog EB1089 Restricts Adrenocortical Tumor Proliferation in NCI-H295 Xenograft Model

The vitamin D (VD) receptor (VDR) is underexpressed in adult and pediatric adrenocortical tumors (ACT), especially in carcinomas (ACC). VDR activation by VD represses B-catenin signaling in different types of tumors that like ACT, present aberrant B-catenin expression. EB1089 is a potent synthetic VD analog that presents lower calcemic effects and is well tolerated at therapeutic doses. Since there is no effective adjuvant target-therapy for patients with ACT, we evaluated the effects of EB1089 in adrenocortical tumor proliferation. Methods:In vitro, we treated NCI-H295 (B-catenin mutant/activated) and Y1 (normal B-catenin) ACC cells with EB1089 (10-7M) and evaluated VDR and B-catenin signaling (qPCR) and cell proliferation (MTS). In vivo, we bilaterally inoculated subcutaneously 10 female NGS mice (8 week-old) with NCI-H295 cells (2.5x106 / 100 mcL). We assessed body weight (BW) and tumor volume (TuV; d2D/2) weekly. Once the TuV reached 100 mm3, we randomized the animals for EB1089 (n=5; 0.8mcg/kg BW) or vehicle [Control group (CG) n=5; 80% propylene glycol-20% PBS] treatment (sc injection; 5 days/week during 5 weeks). At the end of the protocol, mice were sacrificed and each tumor excised, weighted and fixed for pathological evaluation (H&E). Blood samples were taken for serum calcium (colorimetry), cortisol and testosterone (RIA) assessment. Data were presented as percentage or mean ± SEM. Results: In NCI-H295 cells the treatment with EB1089 (48h) increased VDR mRNA expression (47%; p=0.01) and activated its target gene CYP24A1 (p<0.0001). B-catenin expression and signaling decreased, as shown by reduced CTNNB1 (-24%; p=0.001), MYC (-40%; p=0.0001), CCND1 (-38%; p=0.02), and DKK3 (-40%; p=0.02) mRNA expression. Cell proliferation reduced after 96h (-15%; p=0.001). None of these observations were present in Y1 cells. In vivo, EB1089 significantly restricted tumor growth starting after 2 weeks of treatment (TuV: 284 ± 52 vs. 156 ± 20 mm3; p=0.05). At the end of the experiment TuV was ~50% lower in EB1089 xenografts (683 ± 345 vs. 368 ± 211 mm3; p=0.03). TuV and tumor weight correlated positively (r=0.88; p<0.0001). Both, EB1089 and CG tumors presented highly proliferative areas (pleomorphism, mitosis and few apoptotic bodies). However, EB1089 xenografts presented large necrotic areas. BW (p=0.98) and calcium levels (9.96 ± 0.4 vs. 11.3 ± 0.3 mg/dL; p=0.06) did not differ between groups. EB1089 mice presented reduced serum testosterone (76.7 ± 11.6 vs. 39 ± 5.6 ng/dL; p=0.02) and undetectable cortisol levels (CG: 2 ± 1.2 mcg/dL). Conclusion: The activation of VDR signaling by EB1089 inhibited B-catenin activated ACT proliferation. Furthermore, in vivo, EB1089 reduced tumor steroid secretion and did not result in important VD side effects like significant hypercalcemia and weight loss. VD analogs have antitumor activity and may emerge as an adjuvant therapy for patients with ACT.

FSH therapy for idiopathic male infertility: four schemes are better than one

Objective The aim of this article is to propose an algorithm that aids the clinician to choose the best therapeutic scheme of follicle-stimulating hormone (FSH) in the treatment of men with idiopathic infertility, based on testicular volume (TV) and serum total testosterone concentrations; highlighting the potential role of additional therapy with hCG in a sequential temporal scheme. Materials and methods We subdivided patients in four clinical groups: patients with normal TV and serum testosterone concentrations (A); patients with normal TV and reduced serum testosterone concentrations (B); patients with reduced TV and serum testosterone concentration (C); patient with low TV e normal serum testosterone concentrations (D). Then, we administered to each group a specific therapeutic scheme. Group A: treated with FSH alone for at least 3 months; group B: treated with hCG alone twice a week for 3 months and addition of FSH for poor responders (unmodified sperm parameters); group C: treated ab initio with FSH and hCG until the pregnancy was reached; group D: treated with FSH alone for 3 months and addition of hCG for moderate poor responders (increased TV but unmodified sperm parameters) or second cycle of FSH for 3 months for severe poor responders (unmodified TV and sperm parameters). After 6 months we evaluated the therapeutic response in term of sperm parameters normalization rate, spontaneous pregnancy rate, and sperm DNA fragmentation normalization rate. Results 40% of patients became normozoospermic after treatment, while 30% achieved spontaneous pregnancy. B was the group that best responded to treatment in terms of normalization of seminal parameters; while the highest spontaneous pregnancy rate was obtained from the D group. B group also obtained the highest sperm DNA fragmentation normalization rate. Conclusions To date, no reliable predictors of response to treatment with FSH exist, but TV and serum testosterone concentrations can help the clinician to choose the best therapeutic scheme for men with idiopathic infertility. The groups treated with a sequential temporal scheme (B and D groups) showed better clinical results compared with two groups treated with conventional schemes (A and C groups).

Stem Leydig cell regeneration in the adult rat testis is inhibited after a short-term triphenyltin exposure

Triphenyltin (TPT) is an organotin compound and may be an endocrine disruptor, impairing the male reproductive system. However, the effect of short-term TPT exposure on stem Leydig cell regeneration later on remains unknown. Here, we show that TPT affects stem Leydig cell regeneration in the adult rat testis. Adult male Sprague Dawley rats were gavaged with TPT (0, 0.5, 1.0, 2.0 mg/kg body weight/day) for 10 days, followed by a single intraperitoneal injection of ethane dimethane sulfonate (EDS, 75 mg/kg body weight) to eliminate Leydig cells. Testis parameters and hormone levels were investigated on post-EDS days 21, 35, and 56. TPT significantly reduced serum testosterone levels, decreased Leydig cell number and cell size, and down-regulated its specific gene and protein expression at 1.0 and 2.0 mg/kg even 56 days after cession of treatment. TPT lowered PCNA-labeling index of progenitor Leydig cells on post-EDS day 21. TPT also lowered AKT1 and AKT2, and ERK1/2 phosphorylation on post-EDS day 56. This study reveals that a short-term exposure to TPT blocks stem Leydig cell regeneration in the long term thus delaying spermatogenesis.

Dibutyltin Dichloride Retards Leydig Cell Developmental Regeneration in Adult Rat Testis.

Dibutyltin dichloride (DBTCl), widely used as plastic stabilizer, can cause comprehensive toxicity. The present study aims to investigate the effects of DBTCl on rat Leydig cell developmental regeneration and characterize the related mechanism. Adult male Sprague Dawley rats were randomly divided into four groups and gavaged with saline (control) or 5, 10, or 20 mg/kg/day of DBTCl consecutively for 10 days. At the end of the DBTCl treatment, all rats received a single intraperitoneal injection (i.p.,) of 75 mg/kg ethane dimethane sulfonate (EDS) to eliminate all the adult Leydig cells and to induce Leydig cell developmental regeneration. Leydig cell developmental regeneration was evaluated by measuring the levels of serum testosterone, luteinizing hormone, and follicle-stimulating hormone on days 7, 35, and 56 post-EDS. Leydig cell gene and protein expression levels, as well as cell morphology and cell counts were also carried out on day 56 post-EDS. The present study found that DBTCl significantly reduced serum testosterone levels on days 35 and 56 post-EDS, but increased serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels on day 56 at ≥ 5 mg/kg/day. The mRNA and protein levels of Leydig (Lhcgr, Scarb1, Star, Cyp11a1, Hsd17b3, and Hsd11b1) and Sertoli cells (Fshr, Amh, and Sox9) were significantly downregulated in the DBTCl-treated testes compared to the control. Immunohistochemical staining showed that DBTCl-treatment caused fewer regenerated Leydig cells and impaired Sertoli cell development and function in the testis on day 56 post-EDS. In conclusion, the present study demonstrates that DBTCl retards rat Leydig cell developmental regeneration by downregulating steroidogenesis-related enzymes at the gene and protein levels, inhibiting Leydig cell proliferation and impairing Sertoli cell function and development.

Oncostatin M inhibits differentiation of rat stem Leydig cells invivo and invitro.

Oncostatin M (OSM) is a pleiotropic cytokine within the interleukin six family of cytokines, which regulate cell growth and differentiation in a wide variety of biological systems. However, its action and underlying mechanisms on stem Leydig cell development are unclear. The objective of the present study was to investigate whether OSM affects the proliferation and differentiation of rat stem Leydig cells. We used a Leydig cell regeneration model in rat testis and a unique seminiferous tubule culture system after ethane dimethane sulfonate (EDS) treatment to assess the ability of OSM in the regulation of proliferation and differentiation of rat stem Leydig cells. Intratesticular injection of OSM (10 and 100 ng/testis) from post‐EDS day 14 to 28 blocked the regeneration of Leydig cells by reducing serum testosterone levels without affecting serum luteinizing hormone and follicle‐stimulating hormone levels. It also decreased the levels of Leydig cell‐specific mRNAs (Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1 and Hsd11b1) and their proteins by the RNA‐Seq and Western blotting analysis. OSM had no effect on the proliferative capacity of Leydig cells in vivo. In the seminiferous tubule culture system, OSM (0.1, 1, 10 and 100 ng/mL) inhibited the differentiation of stem Leydig cells by reducing medium testosterone levels and downregulating the expression of Leydig cell‐specific genes (Lhcgr, Star, Cyp11a1, Hsd3b1, Cyp17a1 and Hsd11b1) and their proteins. OSM‐mediated action was reversed by S3I‐201 (a STAT3 antagonist) or filgotinib (a JAK1 inhibitor). These data suggest that OSM is an inhibitory factor of rat stem Leydig cell development.

Impaired sleep is associated with low testosterone in US adult males: results from the National Health and Nutrition Examination Survey

Testosterone deficiency has been linked to several adverse health outcomes and recent data have suggested that abnormal sleep quality may result in lower testosterone levels. We assessed the effect of self-reported sleep patterns on serum testosterone while controlling for co-morbidities, and baseline demographics. Using data collected from the 2011-2012 National Health and Nutrition Examination Survey (NHANES), we extracted serum total testosterone level, sleep duration, demographic, and co-morbidities for men aged 16years and older. Univariate and multivariate linear regression was used to estimate the association of number of hours slept, co-morbidities, and demographics with serum testosterone. Among the 9756 individuals in the NHANES dataset, 2295 (23.5%) were males 16years and older with a median (interquartile range) age of 46years (29-62) who also had serum testosterone levels drawn. Median serum testosterone level was 377ng/dL (IQR: 279-492ng/dL). Median number of hours slept was 7h (IQR: 6-8h). On multivariate linear regression, we found serum testosterone decreased by 0.49ng/dL per year of age (p = 0.04), 5.85ng/dL per hour loss of sleep (p < 0.01) and 6.18ng/dL per unit of body mass index (BMI) increase (p < 0.01). Among men aged 16-80 in the United States, we found increasing age, impaired sleep and elevated BMI is associated with low testosterone. It is important, therefore, that evaluation and treatment of reduced serum testosterone should also include improving sleep duration in combination with weight management.

CBLB502, a Toll-like receptor 5 agonist, offers protection against radiation-induced male reproductive system damage in mice.

CBLB502, a Toll-like receptor (TLR)5 agonist derived from Salmonella flagellin, was shown to protect mammalian hematopoietic and gastrointestinal systems from acute irradiation syndrome and to stimulate regeneration. To explore whether CBLB502 can improve testicular injuries caused by irradiation, mice were intraperitoneally injected with 0.2 mg/kg CBLB502 or vehicle control 30 min prior to applying 5.0 Gy ionizing radiation (IR). We observed these mice for the following 120 days and determined that CBLB502 pretreatment alleviated IR-induced oxidative stress, alleviated the distorted architecture of seminiferous tubules, reversed the decline of sperm quantity and quality, and helped recover male mouse fertility. Additionally, CBLB502 efficiently reduced DNA damage and chromosomal aberrations in IR-treated mice and their offspring. Due to the suppression of p53-dependent apoptosis, in IR-treated mice, CBLB502 was shown to significantly activate the nuclear factor kappa B (NFκB) pathway and reduce the apoptotic rate in association with an increase in anti-apoptotic B-cell lymphoma 2 levels and a decrease in the levels of DNA repair protein and proliferating cell nuclear antigen. Moreover, an IR-induced reduction in serum testosterone and superoxide dismutase levels and an increase in malondialdehyde levels were considerably reversed in CBLB502-pretreated mice. No significant reverse effects were found in Tlr5 knockout mice, suggesting that protection of the testis against IR by CBLB502 is primarily dependent on the TLR5 signaling pathway. Our results may help further investigations into potential CBLB502 applications for the protection of the male reproductive system during radiotherapy.

Protective Effects of α-lipoic acid on Biological changes Induced by α- cypermethrin in Testis Rats

Abstract a-cypermethrin is one of the most potent insecticides used worldwide. This study was planned to evaluate the possible role of a-lipoic acid in a-cypermethrin induced toxicity in rats. The treated groups were; the control, α-cypermethrin, a-lipoic acid and a-cypermethrin, and a-lipoic acid groups. Our results showed that administration of a-cypermethrin caused a significant decrease in RBC count, PCV, and Hb content and an increase in WBC count. Also, a- cypermethrin caused a significant increase in the levels of cholesterol, TGs, LDL-Cand VLDL-C, while the HDL-C was decreased. Also, a-cypermethrin caused a reduction in serum testosterone, FSH, and LH levels in intoxicated rats. Furthermore, the co-administration of a-lipoic acid mitigated the toxicity of a-cypermethrin by partially normalizing these biochemical parameters. Our results were supported by histopathological observations of the testis. Our data suggest that a-lipoic acid may have a protective role against a-cypermethrin induced toxicity in rats.

12-week treadmill exercise program elicits lower energy availability without changes in serum testosterone in male rats

The purpose of this study was to investigate whether a treadmill endurance exercise program would reduce serum testosterone and leptin in male rats and assess the impact of increased dietary cholesterol on serum hormones. Male Sprague-Dawley Rats (n = 20) were randomly assigned to a control group (C) or an exercise training group (EX) that performed treadmill running 40 min/day, 6 days/week for 12 weeks. At study midpoint (week 6), rats were randomized to a high-cholesterol (HC) diet (n = 10) or remain on standard semi-purified (LC) diet (n = 10). Results are presented as median [IQR]. At end of week 6, EX + LC had significantly lower body weight (508 [460–527] vs 570 [516–606] g; p = 0.01), mean daily energy intake (76.3 [74.9–82.2] vs 90.9 [86.9–94.5] kcal; p < 0.01), and serum leptin (0.4 [0.3–0.6] vs 3.3 [2.0–4.0] ng/mL; p < 0.01) in comparison to C + LC. No difference was observed between EX + LC and C + LC in serum testosterone (12.6 [6.9–18.3] vs 11.7 [7.6–18.9] ng/mL). At end of week 12, EX + LC had significantly lower body weight (514 [483–568] g) compared to C + LC (644 [575–680] g; p < 0.01) and C + HC (650 [583–702] g; p < 0.01). Serum Leptin in both EX + LC (0.6 [0.2–0.8] ng/mL) and EX + HC (0.6 [0.3–1.6] ng/mL) was significantly lower than C + LC (3.0 [2.2–4.2] ng/mL; p < 0.01). No significant difference in testosterone was observed between C + LC and C + HC (4.3 [3.3–8.3] vs 6.3 [3.2–9.3] ng/mL, respectively). Despite lower energy availability, exercise-induced changes in sex hormones may not occur in training programs ≤ 12 weeks. Lower voluntary energy intake observed in exercise groups despite greater energy expenditure may indicate that lower energy availability in endurance-trained individuals may be inadvertent.

Autophagy regulates testosterone synthesis by facilitating cholesterol uptake in Leydig cells.

Testosterone is indispensable for sexual development and maintaining male characteristics, and deficiency of this hormone results in primary or late-onset hypogonadism (LOH). Testosterone is primarily produced in Leydig cells, where autophagy has been reported to be extremely active. However, the functional role of autophagy in testosterone synthesis remains unknown. In this study, we show that steroidogenic cell-specific disruption of autophagy influenced the sexual behavior of aging male mice because of a reduction in serum testosterone, which is similar to the symptoms of LOH. The decline in testosterone was caused mainly by a defect in cholesterol uptake in autophagy-deficient Leydig cells. Further studies revealed that once autophagic flux was disrupted, Na+/H+ exchanger regulatory factor 2 (NHERF2) accumulated in Leydig cells, resulting in the down-regulation of scavenger receptor class B, type I (SR-BI) and eventually leading to insufficient cholesterol supply. Collectively, these results reveal that autophagy promotes cholesterol uptake into Leydig cells by eliminating NHERF2, suggesting that dysfunction of autophagy might be causal in the loss of testosterone production in some patients.