Testosterone Production Research Articles

Lower malathion concentrations reduce testosterone biosynthesis by Leydig TM3 cells in vitro by altering cellular redox profile and inducing oxidative damage

Malathion is an organophosphate pesticide used in agriculture and control of the Aedes aegypti mosquito. As previous reports have indicated the potential of malathion to compromise testosterone production in in vivo models, the objective of this study was to elucidate the mechanisms underlying the impairment of Leydig cell function, considering its critical role in male reproductive function. To this end, murine Leydig TM3 cells were exposed to concentrations of 1, 10, 100 or 1000 μM malathion for 24 h for evaluation of the compound on cell viability. Subsequently, concentrations of 1, 10, and 100 μM malathion were employed for a 24-h period to assess testosterone biosynthesis, levels of cytokines IL-1β, IL-6, IL-10, and TNF-α, as well as the redox profile. Malathion exerted a concentration-dependent impact on cell viability. Notably, the lower concentrations of malathion (1 and 10 μM) were found to impair testosterone biosynthesis in TM3 cells. While there were changes in IL-1 and TNF-α levels at specific concentrations, no direct correlation with altered hormone production was established. Our investigation revealed that varied malathion concentrations induced oxidative stress by increase in superoxide anion and a compensatory rise in antioxidants. In conclusion, the observed changes in the oxidative profile of TM3 cells were linked to functional impairment, evidenced by reduced testosterone biosynthesis at lower malathion concentrations.

An intracellular VHH targeting the Luteinizing Hormone receptor modulates G protein-dependent signaling and steroidogenesis

Luteinizing hormone (LH) is essential for reproduction, controlling ovulation and steroidogenesis. Its receptor (LHR) recruits various transducers leading to the activation of a complex signaling network. We recently identified iPRC1, the first variable fragment from heavy-chain-only antibody (VHH) interacting with intracellular loop 3 (ICL3) of the follicle-stimulating hormone receptor (FSHR). Because of the high sequence similarity of the human FSHR and LHR (LHCGR), here we examined the ability of the iPRC1 intra-VHH to modulate LHCGR activity. In this study, we demonstrated that iPRC1 binds LHCGR, to a greater extent when the receptor was stimulated by the hormone. In addition, it decreased LH-induced cAMP production, cAMP-responsive element-dependent transcription, progesterone and testosterone production. These impairments are not due to Gs nor β-arrestin recruitment to the LHCGR. Consequently, iPRC1 is the first intra-VHH to bind and modulate LHCGR biological activity, including steroidogenesis. It should help further understand signaling mechanisms elicited at this receptor and their outcomes on reproduction.

Role of p57KIP2 in Stem and Progenitor Leydig Cells of Mouse Testes.

Precise control of proliferation and differentiation of Leydig cells is important for gonadal androgenesis and spermatogenesis. Though cyclin-dependent kinase inhibitors are crucial for cell proliferation and differentiation, their role in the development of early adult Leydig cells (ALCs) remained unanswered. To understand mechanism for ALC development, functional expression of p57KIP2 (cdkn1c) was investigated in the stem Leydig cells (SLCs) and progenitor Leydig cells (PLCs) in mice. The roles of p57KIP2 in the proliferation, differentiation, apoptosis, and steroidogenesis in SLCs and PLCs were investigated by antibodies and bromodeoxyuridine (BrdU) labeling in the early neonatal testes and p57kip2 siRNA in the isolated SLCs and PLCs. Steroidogenic differentiation of PLCs was examined by progesterone and testosterone production in cell culture. From postnatal day (PND) 1 to 14, p57KIP2(+) spindle-shaped cells in the testis interstitium were α-smooth muscle actin (αSMA)(-), a peritubular myoid cells marker, suggesting that they are SLCs and PLCs. Besides, p57KIP2 was also expressed in HSD3β(+) fetal Leydig cells. From PND1 to 14, BrdU(+)/αSMA(-), Ki67(+)/p57KIP2(+), and BrdU(+)/p57KIP2(+) spindle-shaped cells were gradually decreased. From PND1 to 14, p57KIP in the αSMA(-)/p57KIP2(+) cells was peaked at PND7 and decreased thereafter. In THY1(+) isolated SLCs, p57kip2 siRNA significantly increased ki67 and pcna mRNA and pdgfrα mRNA, a differentiation marker and decreased nestin mRNA, a SLC marker. No significant difference in apoptosis related genes mRNA was found after p57kip2 siRNA treatment. In HSD3β(+) PLCs, p57kip2 siRNA increased proapoptotic genes mRNA, annexin V(+) early-apoptotic cells. Importantly, p57kip2 siRNA significantly decreased hsd3β6 and cyp17a1 mRNA and progesterone production. p57KIP2 may suppress proliferation and support stemness of SLCs. In PLCs, p57KIP2 may suppress apoptosis and potentiate the steroidogenic differentiation.

Response of Male Reproductive System Against Ionizing Radiation and Available Radio-protective Agents: Cellular and Molecular Insight.

The reproductive organ, housing spermatogonial stem cells (SSCs), undergoes ongoing division impacted by the irradiation dosage and exposure duration. Within the male reproductive organ, germ stem cells (spermatogonia) and somatic cells (Sertoli and Leydig cells) are present. Lower doses of ionizing (>4-6 Gy) and non-ionizing radiation (radiofrequency and microwave range 900 MHz - 2.45 GHz) may cause sperm-related issues, while higher doses (15 Gy) may affect Leydig cells and testosterone production. Response to radiation varies with age and pubescence. Spermatogonial stem cells, crucial for regenerating the spermatogenic lineage, express molecular markers like Estrogen receptor, FSH (Follicular Stimulating Hormone) receptor, TLR-4 (Toll-like Receptor-4), TLR-5 (Toll-like Receptor-5), FGF2 (Fibroblast Growth Factor-2), KIT (Receptor Tyrosine Kinase), AT-1 (Angiotensin II Type-1 Receptor), LXRs-γ (Liver X Receptor-γ), TNF-β (Tumor Necrosis Factor-β), and PCNA (Proliferating Cell Nuclear Antigen), influencing stem cell activity in testes. This study aimed to review the various available radioprotective agents and their efficacy in targeting the male reproductive system from the available literature. Various radioprotective herbal/synthetic/microbial/metallic extracts/formulations/ drugs [Septilin, Silymarin, Organic Turmeric, Oestrogen, Melatonin, Febuxostat, SQGD (Semiquinone glucoside derivative), Rapamycin, Entolimod, Zinc, Selenium, etc.] have been investigated up to exposure, but owing to effectiveness issues, they are unable to fulfil the aim to the fullest of restoring male fertility and normal testosterone levels during such eventuality. Further study is needed to optimize these tactics and fill knowledge gaps. Also, the effective components of herbal, synthetic drugs, etc., should be isolated and tested up to clinical levels, paving the way for successful radioprotection and radiomitigation strategies in the male reproductive system.

Andropause: A Neglected Disease Entity

This review aims to bring attention to the growing issue of andropause, a condition that affects men as they age. The literature was identified by searching the major bibliographic databases, including PubMed, Google Scholar, EBSCOhost, Web of Science, and CINAHL. Studies were included if they focused on strategies for reducing and preventing andropause, published between 1980 and 2023. Andropause, which is often overlooked and misconceived, is the result of a decrease in testosterone production and can result in physical and psychological symptoms such as fatigue, weight gain, decreased libido, and depression. This paper reviews the literature on the prevalence, detection, and management of andropause. It is important for the medical community to recognize andropause, as diagnosing and treating the condition can significantly improve the quality of life. The review canvases several studies to understand andropause comprehensively, highlighting various biomarkers and diagnostic criteria, possible therapies, and potential risks and complications associated with the condition. In addition, this review offers practical suggestions to health professionals, helping them recognize patients at risk of andropause, assess patients for the condition, and provide appropriate treatments. Furthermore, it emphasizes the importance of regular screening and active monitoring for any possible early warning signs of andropause. This review provides a comprehensive overview of andropause, from identification to management. It demonstrates the need for increased awareness and acknowledgment of andropause among medical professionals in order to ensure that all men get the support they need throughout the aging process.

Estrogenic endocrine disruptor exposure directly impacts erectile function

Erectile dysfunction (ED) is an extremely prevalent condition which significantly impacts quality of life. The rapid increase of ED in recent decades suggests the existence of unidentified environmental risk factors contributing to this condition. Endocrine Disrupting Chemicals (EDCs) are one likely candidate, given that development and function of the erectile tissues are hormonally dependent. We use the estrogenic-EDC diethylstilbestrol (DES) to model how widespread estrogenic-EDC exposure may impact erectile function in humans. Here we show that male mice chronically exposed to DES exhibit abnormal contractility of the erectile tissue, indicative of ED. The treatment did not affect systemic testosterone production yet significantly increased estrogen receptor α (Esr1) expression in the primary erectile tissue, suggesting EDCs directly impact erectile function. In response, we isolated the erectile tissue from mice and briefly incubated them with the estrogenic-EDCs DES or genistein (a phytoestrogen). These acute-direct exposures similarly caused a significant reduction in erectile tissue contractility, again indicative of ED. Overall, these findings demonstrate a direct link between estrogenic EDCs and erectile dysfunction and show that both chronic and acute estrogenic exposures are likely risk factors for this condition.

Effect of Curcuma longa extract on reproduction function in mice and testosterone production in Leydig cells.

Curcuma longa, best known for its culinary application as the main constituent of curry powder, has shown potential impact on the reproductive system. This study aimed to investigate the efficacy of Curcuma longa extract (CLE) on Kidney-Yang deficiency mice induced by hydrocortisone and the possible roles in testosterone secretion in Leydig cells. We evaluated male sexual behaviour, reproductive organ weight, testosterone levels, and histological tissue changes in hydrocortisone-induced mice. CLE effectively reversed hydrocortisone-induced Kidney-Yang deficiency syndrome by improving sexual behaviour, testis and epididymis weight, testosterone levels and reducing pathological damage. Our invitro study further indicated that CLE stimulated testosterone production via upregulating the mRNA and protein expression of steroidogenic enzymes in Leydig cells. It significantly improved H89-inhibited protein expression of StAR and cAMP-response element-binding (CREB), as well as melatonin-suppressed StAR protein expression. The data obtained from this study suggest that CLE could alleviate Kidney-Yang deficiency symptoms and stimulate testosterone production by upregulating the steroidogenic pathway. This research identifies CLE as a potential nutraceutical option for addressing testosterone deficiency diseases.

Gross as well as microscopic anatomy and physiological functions of fetal placenta in Jaffrabadi buffaloes

The present study was conducted to know the gross morphology and histo-morphological structure of fetal placenta in Jaffrabadi buffaloes. Parameters like calf weight, placental weight, numbers, and size of cotyledons were observed in Jaffrabadi buffaloes and morphologically, fetal cotyledons were convex and non-pendunculated in Jaffrabadi buffaloes were found. Histological studies of small and large cotyledons showed extensive branching of secondary and tertiary villi that were longer, slender, and well developed in Jaffrabadi buffalo. A less developed basal lamina was seen in small cotyledon whereas developed basal lamina with numerous capillaries and connective tissue were observed in the large cotyledon. The diameter of trophoblast giant cells (TGC) in larger cotyledons were significantly (P<0.05) than the small cotyledons in expelled placenta at full term in Jaffrabadi buffaloes. A distinct distribution of carbohydrate and lipids in cotyledons were observed between large and small cotyledons as evident by acid mucopolysaccharides, neutral polysaccharides, and sudanophilic staining. Specific staining for calcium with Alizarin red stain showed that calcium is not present in a noticeable amount in small and large cotyledons. Isolation and culture of Jaffrabadi placental cells in M-199 medium with antibiotics and 2% FBS results in the efficient production of progesterone, estrogen, and testosterone. This study has shown that trophoblast cells are the actual sites for steroid hormone production. These cultured placental cells (1x106 cells/ ml) produce Progesterone, Estradiol-17β and Testosterone in the range of 1.72 to 2.12, 16.03 to 19.51 and 0.51 to 0.58 ng/ml, respectively in Jaffrabadi buffalo.

A Systemic Review on Use of Medicinal Plant for Management of Male Infertility

A recent issue with current society is the fall in male infertility. In 50% of instances, couple infertility is caused by the failure in spermatogenesis of the male partner. The male reproductive system is impacted by a variety of factors that lead to infertility, including genitourinary tract infections, endocrine abnormalities, immunological factors, lifestyle, stress, and drug-related damage. There are medications and therapies for male infertility, but they are not very effective, have harmful side effects, and are contraindicated. Plants are among the greatest natural cures for illnesses since they have very few side effects and include a variety of phytoconstituents from different chemical classes. Depending on their concentration, plant bioactive such as flavonoids, polyphenols, glycosides, alkaloids, and terpenoids might have a variable effect on sperm quality - either positively or adversely. These compounds have been suggested as organic reactive oxygen species scavengers for the treatment of male infertility. This review makes an effort to compile the available information and offer perspectives for upcoming investigations on how plants affect male gonads. This review summarizes the most recent research on the use of phytoconstituents to increase testosterone production, support healthy spermatogenesis, and prevent age-related degenerative illnesses linked to low levels of testosterone. This review includes information on about 45 herbal plants that have been published in several papers. The articles have several headings, including plant taxonomy, bioactive components, impacts on fertility, and medical applications. This review provides information on the applications of natural sources for male fertility enhancement and their potential mechanisms of action.

IMPACT OF ANAEROBIC EXERCISES ON EMOTIONAL STATUS

Background. Physical activity can affect a person's mental and emotional well-being, and it can also be an effective way to reduce stress and anxiety. The importance of current topic is also exacerbated by modern lifestyles, which may include more and more stressors. Individual response to anaerobic exercise may vary depending on physical condition, duration and intensity of training, as well as special characteristics. Therefore, understanding how to maintain balance and maintain a healthy approach to sports remains extremely relevant for many people.
 Aim: to analyze the impact of anaerobic exercises on the emotional state.
 Material and methods. The review included 84 articles, which have been chosen using following keywords: «anaerobic exercise», «emotional state», «stress», «cortisol», «testosterone», in PubMed, Scopus and Web of Science databases. Analysis of scientific data has been conducted in order to collect the existed results of researches about the effect of anaerobic exercises on the emotional state.
 Results. In the article we collected data about the relationship of anaerobic physical activity and its impact on emotional state. Present results of researches show that anaerobic physical activity can have a significant impact on emotional state. In particular, it's thought to promote the release of endorphins, improve mood and overall emotional well-being. Additionally, anaerobic exercise has been linked to reduced levels of stress hormones like cortisol. By engaging in intense physical activity, individuals can experience a reduction in feelings of stress and anxiety, as well as an increased ability to cope with difficult situations. Some studies even suggest that anaerobic activity may serve as an effective intervention in the treatment of mood disorders such as depression and anxiety and improves overall quality of life.
 Conclusion. Anaerobic exercise activates sympathomedullary system, hypothalamic-pituitary-adrenal system and testosterone production resulting in positive physiological adaptation. Anaerobic exercise improves mood during and after exercise due to increased synaptic transmission of monoamines, which probably act on the same principle as antidepressant drugs. Physical activity stimulates the release of beta-endorphins which are responsible for feelings of calmness and improved mood after exercise.

Adrenal-Permissive Germline HSD3B1 Allele and Prostate Cancer Outcomes

The adrenal androgen-metabolizing 3β-hydroxysteroid dehydrogenase-1 enzyme, encoded by the HSD3B1 gene, catalyzes the rate-limiting step necessary for synthesizing nontesticular testosterone and dihydrotestosterone production. The common adrenal-permissive HSD3B1(1245C) allele is responsible for encoding the 3β-HSD1 protein with decreased susceptibility to degradation resulting in higher extragonadal androgen synthesis. Retrospective studies have suggested an association of the HSD3B1 adrenal-permissive homozygous genotype with androgen deprivation therapy resistance in prostate cancer. To evaluate differences in mortality outcomes by HSD3B1 genetic status among men with prostate cancer. This cohort study of patients with prostate cancer who were enrolled in the Million Veteran Program within the Veterans Health Administration (VHA) system between 2011 and 2023 collected genotyping and phenotyping information. HSD3B1 genotype status was categorized as AA (homozygous adrenal-restrictive), AC (heterozygous adrenal-restrictive), or CC (homozygous adrenal-permissive). The primary outcome of this study was prostate cancer-specific mortality (PCSM), defined as the time from diagnosis to death from prostate cancer, censored at the date of last VHA follow-up. Secondary outcomes included incidence of metastases and PCSM in predefined subgroups. Of the 5287 participants (median [IQR] age, 69 [64-74] years), 402 (7.6%) had the CC genotype, 1970 (37.3%) had the AC genotype, and 2915 (55.1%) had the AA genotype. Overall, the primary cause of death for 91 patients (1.7%) was prostate cancer. Cumulative incidence of PCSM at 5 years after prostate cancer diagnosis was higher among men with the CC genotype (4.0%; 95% CI, 1.7%-6.2%) compared with the AC genotype (2.1%; 95% CI, 1.3%-2.8%) and AA genotype (1.9%; 95% CI, 1.3%-2.4%) (P = .02). In the 619 patients who developed metastatic disease at any time, the cumulative incidence of PCSM at 5 years was higher among patients with the CC genotype (36.0%; 95% CI, 16.7%-50.8%) compared with the AC genotype (17.9%; 95% CI, 10.5%-24.7%) and AA genotype (18.5%; 95% CI, 12.0%-24.6%) (P = .01). In this cohort study of US veterans undergoing treatment for prostate cancer at the VHA, the HSD3B1 CC genotype was associated with inferior outcomes. The HSD3B1 biomarker may help identify patients who may benefit from therapeutic targeting of 3β-hydroxysteroid dehydrogenase-1 and the androgen-signaling axis.

Immune Checkpoint Inhibitors and Male Fertility: Should Fertility Preservation Options Be Considered before Treatment?

In recent years, immune checkpoint inhibitors (ICIs) have become a viable option for many cancer patients, including specific subgroups of pediatric patients. Despite their efficiency in treating different types of cancer, ICIs are responsible for a number of immune-related adverse events, including inflammatory toxicities, that can affect several organs. However, our knowledge of the impact of ICIs on the testis and male fertility is limited. It is possible that ICI treatment affects testicular function and spermatogenesis either directly or indirectly (or both). Treatment with ICIs may cause increased inflammation and immune cell infiltration within the seminiferous tubules of the testis, disturbing spermatogenesis or testosterone deficiency (primary hypogonadism). Additionally, the interference of ICIs with the hypothalamic-pituitary-gonadal axis may alter testosterone production, affecting testicular function (secondary hypogonadism) and spermatogenesis. This review provides an overview of the available evidence on the potential association between ICIs and the disruption of spermatogenesis, with special focus on ICIs targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed death protein 1 (PD-1) and programmed death-ligand 1 (PD-L1). Moreover, it highlights the need for further investigations and encourages the discussion of associated risks and fertility-preservation considerations between clinicians and patients.

Stem Leydig cells support macrophage immunological homeostasis through mitochondrial transfer in mice

As testicular mesenchymal stromal cells, stem Leydig cells (SLCs) show great promise in the treatment of male hypogonadism. The therapeutic functions of mesenchymal stromal cells are largely determined by their reciprocal regulation by immune responses. However, the immunoregulatory properties of SLCs remain unclear. Here, we observe that SLCs transplantation restore male fertility and testosterone production in an ischemia‒reperfusion injury mouse model. SLCs prevent inflammatory cascades through mitochondrial transfer to macrophages. Reactive oxygen species (ROS) released from activated macrophages inducing mitochondrial transfer from SLCs to macrophages in a transient receptor potential cation channel subfamily member 7 (TRPM7)-mediated manner. Notably, knockdown of TRPM7 in transplanted SLCs compromised therapeutic outcomes in both testicular ischemia‒reperfusion and testicular aging mouse models. These findings reveal a new mechanism of SLCs transplantation that may contribute to preserve testis function in male patients with hypogonadism related to immune disorders.

Association between Fish Intake and Serum Testosterone Levels in Older Males: The Hitachi Health Study II

BackgroundFish are rich in omega-3 polyunsaturated fatty acids and vitamin D, which can promote testosterone synthesis and secretion. However, some contaminants present in fish may disrupt testosterone production. ObjectiveThis study aimed to investigate the association between fish intake (total, fatty, and lean fish) and serum testosterone levels in older males. MethodsThis cross-sectional study included 1545 Japanese males aged 60–69 y who participated in the baseline survey of the Hitachi Health Study II. Fish intake was estimated using a validated brief-type self-administered diet history questionnaire. Total testosterone levels were measured by chemiluminescence immunoassay. Multivariable linear regression analysis was used to analyze the association between fish intake and serum testosterone levels. ResultsHigher total fish intake was associated with higher levels of serum testosterone, with an adjusted mean [95% confidence interval (CI) of 5.63 (5.43, 5.83) and 5.99 (5.78, 6.20)] ng/mL for the 1st and 4th quartiles of total fish intake, respectively (P for trend = 0.06). When analyzing fatty and lean fish separately, higher intake of lean fish, but not fatty fish, was associated with higher levels of serum testosterone: adjusted mean (95% CI): 5.63 (5.43, 5.82) and 6.00 (5.79, 6.20) ng/mL for the 1st and 4th quartiles of lean fish intake, respectively (P for trend = 0.01). ConclusionsAmong older males, higher intake of total fish, particularly lean fish, was associated with higher serum testosterone levels. Curr Dev Nutr 20xx;x:xx.

(118) Treatment Postcovid Hypogonadism with Autologous Stem Cells and PRP

Abstract Introduction COVID-19 affects ACE receptors, causing autoimmune processes that damage testicular tissue, disrupting the processes of spermatogenesis and testosterone production. Recently, we have seen an increasing number of patients with erectile dysfunction ED appearing 3–6 month safter COVID-19 and prolonging for years. Objective The aim of our study was to determine effectiveness of hormone stimulating and replacement therapy, possibility of restorative therapy with autologous stem cells (ASC) and Plasma rich Platelets (PRP) –therapy in patients with hypogonadism. Methods 117 patients with ED, which appeared 3–6 months after COVID-19. All patients except ED had clinical signs of decreased testosterone: decreased libido, mental and cognitive functions, decreased performance, fatigue. Testosterone fractions (total and free), LH, Estradiol, FSH before, during and after rehabilitation therapy were studied. Patients were divided into 2 groups: 1st- 68 patients with real hypogonadism (Tm = 7.43 + 2.1 below 12 nmol / l), and 2nd - 49 patients with relative hypogonadism (Tm = 14.1 + 1.8 above 12 nmol / l). l), they were studied the sensitivity of tissue receptors to testosterone. Results First group - patients were prescribed Tribulus terrestris extract 45% - 750 mg, Fenugreek extract 50% - 25 mg twice a day for 2 months. 37 patients (54.4%), group 1a (mean age 41.6 + 4.3) responded to therapy by normalizing the level of T (more than 12 nmol/l) and improving erectile function, 31 patients (45.6%), group 1b (average age 47.3 + 5.2) without the effect of herbal medicine prescribed testosterone Omnadren 250 1 time in 3 weeks. After 2 months of hormone replacement therapy, 28 patients were diagnosed with normal T levels, but only 19 patients (61.2%) in group 1b improved their erectile function. Second group - 49 patients were also prescribed Tribulus terrestris extract 45% - 750 mg, Fenugreek extract 50% - 25 mg twice a day for 2 months and PRP therapy (intracavernous injections of 2 ml of autoplasma with a platelet concentration greater than 1 million/ml) 1 time in 10 days 8 times. 23 patients - group 2a responded to therapy with the disappearance of symptoms of hypogonadism and improvement of ED. 26 patients - group 2b – did not have significant improvements, so they received 2 courses of 5 intravenous injections of 5 million autologous stem cells (50 millionin total). 17 patients (65.4% of group 2b) improved their mental health, reduction of symptoms of hypogonadism, improvement of erectile function to normal values (IIEF -5 increased to 19.4 + 2.2). Conclusions 1. Multicenter studies of the effect of COVID-19 on fertile and androgenic function in men and the possibility of their recovery in postpartum pathology are still needed and should be prolonged. 2. A significant proportion of men with postcovid hypogonadism and ED are men with normal levels of T, but reduced sensitivity of tissue receptors to T (relative hypogonadism). 3. In the treatment of relative postcovid hypogonadism, intracavernous PRP therapy and intravenous stem cell therapy have a clinical effect and can be considered as promising treatments for this pathology. Disclosure No.

Effect of melatonin on reproductive function in propylthiouracil induced hypothyroidism in adult male rats.

Thyroid hormones are essential for regulating metabolism, reproduction, and growth. Hypothyroidism is connected with lower sperm count and motility, leading to male infertility. Oxidative stress is likely to be linked to this interaction. Melatonin, being known as an oxidative scavenger, may offer a feasible treatment method for reproductive dysfunction accompanying hypothyroidism in adult male rats. The purpose of this investigation was to determine the mechanism by which melatonin treatment affected spermatogenic and steroidogenic function in an experimental model-induced hypothyroidism in adult male rats. Twenty-one male albino adult rats weighing between 150 and 210 g were used in this experiment. Rats were split into three groups and studied for 11 weeks. The control euthyroid group, in which rats received 0.9% Sodium Chloride (NaCl) solution by intraperitoneal injection [solvent for 6-propyl 2-thouracil (PTU)], 6 days/week for 8 weeks; the PTU-induced hypothyroid group, in which chemical thyroidectomy was induced by intraperitoneal injection of PTU at a dose of 10 mg/kg body weight, 6 days/week for 8 weeks; and the melatonin-treated hypothyroid group, which received 3 mg/kg melatonin intraperitoneally daily for 21 days plasma free Triiodothyronine (T3), free Thyroxin (T4), thyroid stimulating hormone (TSH), free testosterone, Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and prolactin were measured. Also, semen analysis, testicular tissue malondialdehyde (MDA), and tumor necrosis factor-α (TNF-α) were accessed. The results indicated that melatonin significantly increased sperm viability and motility compared to the untreated PTU-induced hypothyroid group (p<0.001). Testicular MDA and TNF-α showed a significant decrease in the melatonin-treated hypothyroid group compared with the PTU-induced hypothyroid group (p<0.05). In addition, plasma testosterone levels were significantly increased, accompanied by a significant reduction of plasma prolactin levels compared to the untreated hypothyroid group (p<0.05 for both). Based on the study findings, melatonin could mitigate gonadal dysfunction induced by hypothyroidism by improving several components of semen analysis, such as sperm motility and sperm viability, as well as by enhancing testosterone production focusing on oxidative and inflammatory stress as the underlying mechanisms.

Two Sustained Release Membranes Used in Formulating Low Strength Testosterone Reservoir Transdermal Patches.

Several Testosterone Replacement Therapies exist for hypogonadism, but an in-depth analysis of these products reveals a high dosing frequency and a high drug loading, up to 120 mg for a potent drug like Testosterone. This may lead to patient non-compliance and toxicity problems if used improperly. Androderm® is the only 24-hour marketed transdermal patch. Testopel pellets are another extended-release product given subcutaneously every 3-6 months. But it requires surgical implantation and is associated with severe adverse effects. For this reason, the development of other extended- release testosterone products is needed. The study aims at using different excipients for the exploration of sustained-release patches. Reservoir formulations were prepared using Carbomer 941 and HPMC K100M as the sustained- release polymers plus CoTran™ 9702 and CoTran™ 9712 as the release membranes. The formulated patches were subjected to in vitro dissolution for 24 h. The amount of drug release at each time point was determined using the developed HPLC assay. FDA Similarity Factor, f2, t-test, and ANOVA were applied to all nine-point collected samples. CoTran™ 9712 membrane was found to be a better candidate than CoTran™ 9702. The in vitro release profiles of the four formulations were compared (Carbomer vs. HPMC, and CoTran™ 9702 vs. CoTran™ 9712). CoTran™ 9702 releases testosterone significantly lower. Carbomer and CoTran™ 9712 were the best formulation among all with the highest drug release of 2.16 ± 0.36 mg.

Fetal Leydig cells: What we know and what we don't.

During male fetal development, testosterone plays an essential role in the differentiation and maturation of the male reproductive system. Deficient fetal testosterone production can result in variations of sex differentiation that may cause infertility and even increased tumor incidence later in life. Fetal Leydig cells in the fetal testis are the major androgen source in mammals. Although fetal and adult Leydig cells are similar in their functions, they are two distinct cell types, and therefore, the knowledge of adult Leydig cells cannot be directly applied to understanding fetal Leydig cells. This review summarizes our current knowledge of fetal Leydig cells regarding their cell biology, developmental biology, and androgen production regulation in rodents and human. Fetal Leydig cells are present in basement membrane-enclosed clusters in between testis cords. They originate from the mesonephros mesenchyme and the coelomic epithelium and start to differentiate upon receiving a Desert Hedgehogsignal from Sertoli cells or being released from a NOTCH signal from endothelial cells. Mature fetal Leydig cells produce androgens. Human fetal Leydig cell steroidogenesis is LHCGR (Luteinizing Hormone Chronic Gonadotropin Receptor) dependent, while rodents are not, although other Gαs -protein coupled receptors might be involved in rodent steroidogenesis regulation. Fetal steroidogenesis ceases after sex differentiation is completed, and some fetal Leydig cells dedifferentiate to serve as stem cells for adult testicular cell types. Significant gaps are acknowledged: (1) Why are adult and fetal Leydig cells different? (2) What are bona fide progenitor and fetal Leydig cell markers? (3) Which signaling pathways and transcription factors regulate fetal Leydig cell steroidogenesis? It is critical to discover answers to these questions so that we can understand vulnerable targets in fetal Leydig cells and the mechanisms for androgen production that when disrupted, leads to variations in sex differentiation that range from subtle to complete sex reversal.

Protective Effect of Black Rice Cyanidin-3-Glucoside on Testicular Damage in STZ-Induced Type 1 Diabetic Rats.

Diabetic testicular damage is quite a common and significant complication in diabetic men, which could result in infertility. The natural fertility rate of type 1 diabetes men is only 50% because of testicular damage. This research first aimed to explore the intervention effect of C3G on testicular tissue damage induced by diabetes. Here, a streptozotocin-induced type 1 diabetic rat model was established, and then C3G was administered. After 8 weeks of C3G supplementation, the symptoms of diabetes (e.g., high blood glucose, lower body weight, polydipsia, polyphagia) were relieved, and at the same time that sperm motility and viability increased, sperm abnormality decreased in C3G-treated diabetic rats. Furthermore, the pathological structure of testis was restored; the fibrosis of the testicular interstitial tissue was inhibited; and the LH, FSH, and testosterone levels were all increased in the C3G-treated groups. Testicular oxidative stress was relieved; serum and testicular inflammatory cytokines levels were significantly decreased in C3G-treated groups; levels of Bax, Caspase-3, TGF-β1 and Smad2/3 protein in testis decreased; and the level of Bcl-2 was up-regulated in the C3G-treated groups. A possible mechanism might be that C3G improved antioxidant capacity, relieved oxidative stress, increased anti-inflammatory cytokine expression, and inhibited the apoptosis of spermatogenic cells and testicular fibrosis, thus promoting the production of testosterone and repair of testicular function. In conclusion, this study is the first to reveal that testicular damage could be mitigated by C3G in type 1 diabetic rats. Our results provide a theoretical basis for the application of C3G in male reproductive injury caused by diabetes.

Nitric oxide-induced lipophagic defects contribute to testosterone deficiency in rats with spinal cord injury.

Males with acute spinal cord injury (SCI) frequently exhibit testosterone deficiency and reproductive dysfunction. While such incidence rates are high in chronic patients, the underlying mechanisms remain elusive. Herein, we generated a rat SCI model, which recapitulated complications in human males, including low testosterone levels and spermatogenic disorders. Proteomics analyses showed that the differentially expressed proteins were mostly enriched in lipid metabolism and steroid metabolism and biosynthesis. In SCI rats, we observed that testicular nitric oxide (NO) levels were elevated and lipid droplet-autophagosome co-localization in testicular interstitial cells was decreased. We hypothesized that NO impaired lipophagy in Leydig cells (LCs) to disrupt testosterone biosynthesis and spermatogenesis. As postulated, exogenous NO donor (S-nitroso-N-acetylpenicillamine (SNAP)) treatment markedly raised NO levels and disturbed lipophagy via the AMPK/mTOR/ULK1 pathway, and ultimately impaired testosterone production in mouse LCs. However, such alterations were not fully observed when cells were treated with an endogenous NO donor (L-arginine), suggesting that mouse LCs were devoid of an endogenous NO-production system. Alternatively, activated (M1) macrophages were predominant NO sources, as inducible NO synthase inhibition attenuated lipophagic defects and testosterone insufficiency in LCs in a macrophage-LC co-culture system. In scavenging NO (2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO)) we effectively restored lipophagy and testosterone levels both in vitro and in vivo, and importantly, spermatogenesis in vivo. Autophagy activation by LYN-1604 also promoted lipid degradation and testosterone synthesis. In summary, we showed that NO-disrupted-lipophagy caused testosterone deficiency following SCI, and NO clearance or autophagy activation could be effective in preventing reproductive dysfunction in males with SCI.