Testosterone Synthesis Research Articles

Electron microscopic changes in interstitial endocrinocytes of rats testicles during administration of triptorellin for 365 days

Triptorelin is a gonadotropin-releasing hormone agonist that is a potent inhibitor of testosterone (in men) and estrogen (in women) synthesis and is used to treat advanced prostate cancer. Studies of the mechanisms of regulation and synthesis of testosterone formation in testicular interstitial cells demonstrate multiple endogenous targets that can increase testosterone biosynthesis, which may moderate the effects of testosterone depletion. Triptorelin, a synthetic analog of the neurohormone gonadoliberin, suppresses the expression of the GnRH receptor in the pituitary gland, but does not change the functioning of the pituitary-testicular complex. The purpose of the work is to study the electron microscopic changes in the interstitial endocrinocytes of the testes of rats after the administration of triptorelin for 365 days. The experiment was conducted on 35 sexually mature male white rats. The rats were divided into 2 groups: the control group (I) was injected with a physiological solution, the II group with central deprivation of the synthesis of luteinizing hormone was injected subcutaneously with triptorelin at a dose of 0.3 mg of the active substance per kg of the rat's body weight. The study of the interstitial space in the testicles of white rats showed that long-term administration of triptorelin causes hormonal dysregulation of the hypothalamus-pituitary-testis system, which leads to quantitative and qualitative changes in the endocrine cells of the interstitial space of the testis, which is confirmed by electron microscopic changes in subcellular structures. The maximum effect of triptorelin is determined from the 180th day of observation, which is characterized by an increase in degenerative changes in endocrinocytes, and the detection of Reinke crystals in the cytoplasm of interstitial endocrinocytes from the 270th day of observation.

Exogenous Nucleotides Ameliorate Age-Related Decline in Testosterone in Male Senescence-Accelerated Mouse Prone-8 (SAMP8) Mice by Modulating the Local Renin-Angiotensin System Antioxidant Pathway.

In older men, an age-related decline in testosterone is closely associated with various adverse health outcomes. With the progression of aging, hyperactivation of the local renin-angiotensin system (RAS) and oxidative stress increase in the testis. The regulation of RAS antioxidants may be a target to delay testicular aging and maintain testosterone levels. Exogenous nucleotides (NTs) have anti-aging potential in several systems, but there are no studies of their effects on the reproductive system. In our study, we examined the effects of exogenous NTs on testosterone synthesis and explored possible mechanisms of action. Therefore, senescence-accelerated mouse prone-8 (SAMP8) mice and senescence-accelerated mouse resistant 1 (SAMR1) were used in the experiment, and they were randomly divided into an NTs free group (NTs-F), a normal control group (control), a low-dose NTs group (NTs-L), a middle-dose NTs (NTs-M), a high-dose NTs group (NTs-H) and SAMR1 groups, and the testis of the mice were collected for testing after 9 months of intervention. The results showed that exogenous NTs could increase the testicular organ index in mice during aging, and delayed the age-associated decline in testosterone levels in SAMP8 male mice, possibly by modulating the local RAS antioxidant pathway and reducing oxidative stress to protect the testis. The present study provides new research clues for the development of preventive and therapeutic strategies for related diseases.

FERTILITY FACTORS. D-HORMONE IN PREVENTION OF REPRODUCOPATHIES CAUSED BY BENIGN PROSTATIC HYPERPLASIA (Literature review)

In accordance with the statistical data, the spreading of benign prostatic hyperplasia (BPH) is increasing, because modern stressful lifestyle causes sex hormones disbalances and other dyshormonal conditions which lead to pathologies’ development at younger ages. The pathogenesis of BPH is connected with disturbances of regulation in the hypothalamo-hypophyseal system and with the changes of the regulatory signals intensity which go to the reproductive organs. The central pathogenic links of this pathology’ development are: the declining of efficacy of hypothalamus functioning regulation which carries out on the basis of feedback; the changes of the pituitary gland cells sensitivity to the releasing hormones of hypothalamus and disbalance of sex hormones. In addition to gonadotropins prolactin has a significant impact on the BPH development. Its hypersecretion causes the development of hyperplastic age-related changes in prostate gland. The introduction of dopamine receptors antagonists such as sulpiride causes the increasing of prolactin concentration and declining of gonadotropins releasing. “Sulpiride model” is broadly used in the experimental endocrinology for studying prostate-active drugs and is characterized by sex hormones disbalance' and inflammation processes’ development. The development of the pathology is linked with the disturbance of testosterone synthesis in the testicles, insufficiency of prostate gland functioning and increasing of prolactin and estradiol concentrations which provoke proliferative changes. The androgens disbalance is the main cause of BHP development, therefore, the “Testosterone model” based on testosterone’ and estradiol’ slow releasing implants is broadly used for modeling of urination disturbance in rats which imitates the same condition in elderly men with benign prostatic hyperplasia. The prostatitis, which developed under the condition of BHP, causes the disturbances of spermatogenesis during experimental modeling and in the clinical practice when spermatopathies, sexual disturbances and hypofertility are diagnosed in patients. The traditional BHP therapeutic methods are not always effective and the new therapeutic approaches of hypofertility caused by prostate gland pathologies are developed by investigators. Thus, there are data about correlation between increased vitamin D consumption and decreasing of BHP spreading, between gland volume and cells proliferation in patients with this pathology. Vitamin D level influences the quantity and quality of ejaculate. The deficiency of vitamin D is associated with lower proportion of sex hormones in young men and choriogonin sensitivity of Leydig's cells to be diminished. It has been experimentally shown that cholecalciferol regulates steroid hormones genesis, mitigates the age-related declining of testicles activity, protects the testicles in old animals and the unchanged spermatogenesis in normal males which received vitamin D3 can be considered to be the result of estrogens and androgens balance. The androgens receptors as well as vitamin D receptors have been found in prostate gland in health males and in those with hyperplasia. In addition to its classical function in calcium homeostasis and bones health, vitamin D inhibits tumor growth in different tissues including prostate gland. Vitamin D metabolites have been determined to be anti-inflammatory, proapoptotic and antiproliferative. D-hormone application potentiates the specific properties of prostate gland protectors, recovers reproductive capacity under the condition of experimental chronic prostatitis of inflammatory genesis as well as traumatic. All facts listed above allow to confirm the positive role of D-hormone in the regulation of prostate gland and reproductive function in patients with BHP and recovering of fertility under the condition of this pathology. The literature search for this review has been carried out on databases PubMed, Science Direct, Europa PMC, BMC, MedLine etc.

Loss of NR5A1 in mouse Sertoli cells after sex determination changes cellular identity and induces cell death by anoikis.

To investigate the role of the nuclear receptor NR5A1 in the testis after sex determination, we analyzed mice lacking NR5A1 in Sertoli cells (SCs) from embryonic day (E) 13.5 onwards. Ablation of Nr5a1 impaired the expression of genes characteristic of SC identity (e.g. Sox9 and Amh), caused SC death from E14.5 onwards through a Trp53-independent mechanism related to anoikis, and induced disorganization of the testis cords. Together, these effects caused germ cells to enter meiosis and die. Single-cell RNA-sequencing experiments revealed that NR5A1-deficient SCs changed their molecular identity: some acquired a 'pre-granulosa-like' cell identity, whereas other reverted to a 'supporting progenitor-like' cell identity, most of them being 'intersex' because they expressed both testicular and ovarian genes. Fetal Leydig cells (LCs) did not display significant changes, indicating that SCs are not required beyond E14.5 for their emergence or maintenance. In contrast, adult LCs were absent from postnatal testes. In addition, adult mutant males displayed persistence of Müllerian duct derivatives, decreased anogenital distance and reduced penis length, which could be explained by the loss of AMH and testosterone synthesis due to SC failure.

Evaluation of Endocrine Related Adverse Effects of Non-Endocrine Targeted Pharmaceuticals in Cellular Systems.

Prenatal period is a critical developmental phase that is sensitive to hormonal disruption by natural and/or exogenous hormones. Some pharmaceuticals frequently prescribed and used safely during pregnancy are shown to interact with the developmental programming of fetus, resulting in endocrine-related adverse effects. In this research, we aimed to determine the endocrine disrupting potential of paracetamol, indomethacin, alpha-methyldopa and pantoprazole which are frequently prescribed pharmaceuticals during pregnancy. In vitro aromatase inhibitory, estrogen receptor (ER) agonist/antagonist (E-Screen assay) and hormone biosynthesis modulatory effects (H295R steroidogenesis assay) of the selected pharmaceuticals were evaluated. Furthermore, their effects on viability of MCF-7/BUS and H295R cells were also evaluated by MTT assay. None of the pharmaceuticals affected H295R cell viability. Only indomethacin reduced MCF-7/BUS cell viability at 100µM and 300µM. Among the tested pharmaceuticals, only paracetamol and indomethacin showed aromatase inhibitory activity with IC 50 values of 14.7 x 10 -5 M and 57.6 x 10 -5 M, respectively. Moreover, indomethacin displayed a biphasic ER agonist effect. ER antagonist effects of indomethacin and pantoprazole were confirmed by performing two stepped E-Screen assay. After the partial validation of the H295R steroidogenesis assay with forskolin and prochloraz, the effects of pharmaceuticals on synthesis of testosterone (T) and estradiol (E2) levels were tested. Alpha-methyldopa increased E2 at all tested concentrations and T at 1.48 and 4.4µM. Contrarily other tested pharmaceuticals did not affect steroidogenesis. Present data suggest that all tested pharmaceuticals may have potential endocrine disrupting effect, which should be considered when used in pregnancy.

Expression of the umami taste receptor T1R1/T1R3 in porcine testis of: Function in regulating testosterone synthesis and autophagy in Leydig cells

Testosterone is a vital male hormone responsible for male sexual characteristics. The taste receptor family 1 subunit 3 (T1R3) regulates testosterone synthesis and autophagy in non-taste cells, and the links with the taste receptor family 1 subunit 1 (T1R1) for umami perception. However, little is known about these mechanisms. Thus, we aimed to determine the relationship between the umami taste receptor (T1R1/T1R3) and testosterone synthesis or autophagy in testicular Leydig cells of the Xiang pig. There was a certain proportion of spermatogenic tubular dysplasia in the Xiang pig at puberty, in which autophagy was enhanced, and the testosterone level was increased with a weak expression of T1R3. Silenced T1R3 decreased testosterone level and intracellular cyclic adenosine monophosphate (cAMP) content and inhibited the messenger RNA (mRNA) expression levels of testosterone synthesis enzyme genes [steroidogenic acute regulatory protein (StAR), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (3β-HSD1), cytochrome P450 family 17 subfamily A member 1 (CYP17A1) and hydroxysteroid 17-beta dehydrogenase 3 (17β-HSD3)]. In addition, T1R3 increased the number of acidic autophagy bubbles and upregulated the expression levels of autophagy markers [Microtubule-associated protein 1 A/1B-light chain 3 (LC3) and Beclin-1] in testicular Leydig cells of the Xiang pig. Using an umami tasting agonist (10 mM L-glutamate for 6 h), the activation of T1R1/T1R3 enhanced the testosterone synthesis ability by increasing the intracellular cAMP level and upregulated the expression levels of StAR, 3β-HSD1, CYP17A1 and 17β-HSD3 in Leydig cells. Furthermore, the number of acidic autophagy bubbles decreased in the T1R1/T1R3-activated group with the downregulation of the expression levels of the autophagy markers, including LC3 and Beclin-1. These data suggest that the function of T1R1/T1R3 expressed in testicular Leydig cells of the Xiang pig is related to testosterone synthesis and autophagy.

Early Castration in Horses Does Not Impact Osteoarticular Metabolism

The castration of stallions is traditionally performed after puberty, at around the age of 2 years old. No studies have focused on the effects of early castration on osteoarticular metabolism. Thus, we aimed to compare early castration (3 days after birth) with traditional castration (18 months of age) in horses. Testosterone and estradiol levels were monitored from birth to 33 months in both groups. We quantified the levels of biomarkers of cartilage and bone anabolism (CPII and N-MID) and catabolism (CTX-I and CTX-II), as well as of osteoarthritis (HA and COMP) and inflammation (IL-6 and PGE2). We observed a lack of parallelism between testosterone and estradiol synthesis after birth and during puberty in both groups. The extra-gonadal synthesis of steroids was observed around the 28-month mark, regardless of the castration age. We found the expression of estrogen receptor (ESR1) in cartilage and bone, whereas androgen receptor (AR) expression appeared to be restricted to bone. Nevertheless, with respect to osteoarticular metabolism, steroid hormone deprivation resulting from early castration had no discernable impact on the levels of biomarkers related to bone and cartilage metabolism, nor on those associated with OA and inflammation. Consequently, our research demonstrated that early castration does not disrupt bone and cartilage homeostasis.

Kualitas Sperma pada Laki-Laki Defisiensi Vitamin D di Palembang

Vitamin D deficiency may play some roles in male infertility. Abundant vitamin D receptors exist in male reproductive tracts, implying some significant functions of vitamin D in spermatogenesis and/or testosterone synthesis. This study aims to measure sperm parameters in males with vitamin D deficiency in Palembang, South Sumatra. This study was a conducted as observational, cross-sectional descriptive study on single mother and child hospital in Palembang. Subjects’ serum vitamin D was measured in an accredited external clinical laboratory and sperm samples were collected and examined in same laboratory according to standard protocols. Median age of subject was 30,0 (24-49) years old. Mean serum vitamin D concentration was 21.372 ± 4.202 ng/mL. Median sperm concentration was 21.75 (0.00-137.00) millions/mL. Median sperm count was 66.14 (0.00-370.80) millions. Average progressive motility was 35.64 ± 18.573%. Median normal morphology was 5.00 (0.00-7.00)%. Normal sperm parameters observed in this study implicated vitamin D plays no significant role in sperm quality.

PFOS-elicited metabolic perturbation in liver and fatty acid metabolites in testis of adult mice.

Multiple factors can contribute to sub-fecundity, including genetics, lifestyle, and environmental contaminants. PFASs are characterized as "forever chemicals" due to their ubiquitous contamination and their persistence in the environment, wildlife, and humans. Numerous studies have demonstrated that PFAS exposure adversely affects multiple bodily functions, including liver metabolism and gonadal function. It is unclear, however, how the disruption of hepatic fatty acid metabolism affects testicular function. In this study, male mice were administered 0.3 and 3 μg/g body weight of PFOS for 21 days. Our data showed that PFOS exposure caused hepatic steatosis, as evidenced by significant increases in triglyceride levels, expression of ATP-citrate lyase, and fatty acid synthase, as well as fasting insulin levels. PFOS perturbed the expression levels of hepatokines, of which fibroblast growth factor-21 (Fgf-21), leukocyte cell-derived chemotaxin-2 (Lect-2), and retinol-binding protein-4 (Rbp-4) were significantly reduced, whereas angiopoietin-like 4 (Angptl4) was noticeably increased. While Rbp-4 and Fgf-21 are known to contribute to spermatogenesis and testosterone synthesis. In PFOS-exposed groups, testicular ATP, and testosterone decreased significantly with a significant increase in the expression of peroxisome proliferator-activated receptor-coactivator 1α. Mass spectrophotometry imaging revealed the localization of PFOS in testes, along with significant increases in fatty acid metabolites. These included arachidonic acid, dihomo-α-linolenic acid, dihomo-γ-linolenic acid, oxidized ceramide, diacylglycerol, phosphatidylcholine, and phosphatidylethanolamine, which are associated with inflammation and post-testicular causes of infertility. This study revealed potential links between PFOS-elicited changes in hepatic metabolism and their impacts on testicular biology. This study provides insights into alternative targets elicited by PFOS that can be used to develop diagnostic and therapeutic strategies for improving testicular dysfunction.

In vitro antiandrogenic effects of the herbicide linuron and its metabolites

Although the herbicide linuron is banned for use in the EU due to its reproductive and developmental toxicity, it can still be found in randomly sampled foods grown in and outside the EU. It is not clear if metabolites of linuron can contribute to the endocrine disrupting effects following exposure to the parent compound. To address this gap, we analysed linuron and the metabolites 1-(3,4-dichlorophenyl) urea (DCU), 3,4-dichloroaniline (DCA) and 1-(3,4-dichlorophenyl)-3-methoxyurea (DCXU) for androgen receptor (AR) activities and effects on steroidogenesis. Generally, linuron and the metabolites showed qualitatively similar antiandrogenic profiles, but potencies varied. All compounds were AR antagonists, with linuron showing highest potency (IC50 of 2.8 μM). The overall picture of effects on steroidogenesis showed that linuron and metabolites increased the levels of estrogens and corticosteroids, whereas the synthesis of androgens was inhibited. The metabolite DCU was by far the most potent inhibitor of testosterone synthesis (IC50 of 6.7 μM compared to IC50 of 51.1 μM for linuron). We suggest that it is likely that the metabolites contribute to the antiandrogenic effects of linuron in vivo, especially by inhibiting testosterone synthesis.

Melatonin Inhibits Testosterone Synthesis in Rooster Leydig Cells by Targeting CXCL14 through miR-7481-3p

Melatonin has been proved to be involved in testosterone synthesis, but whether melatonin participates in testosterone synthesis by regulating miRNA in Leydig cells is still unclear. The purpose of this study is to clarify the mechanism of melatonin on Leydig cells testosterone synthesis from the perspective of miRNA. Our results showed that melatonin could significantly inhibit testosterone synthesis in rooster Leydig cells. miR-7481-3p and CXCL14 were selected as the target of melatonin based on RNA-seq and miRNA sequencing. The results of dual-luciferase reporter assays showed that miR-7481-3p targeted the 3′-UTR of CXCL14. The overexpression of miR-7481-3p significantly inhibited the expression of CXCL14 and restored the inhibitory role of melatonin testosterone synthesis and the expression of StAR, CYP11A1, and 3β-HSD in rooster Leydig cells. Similarly, interference with CXCL14 could reverse the inhibitory effect of melatonin on the level of testosterone synthesis and the expression of StAR, CYP11A1, and 3β-HSD in rooster Leydig cells. The RNA-seq results showed that melatonin could activate the PI3K/AKT signal pathway. Interference with CXCL14 significantly inhibited the phosphorylation level of PI3K and AKT, and the inhibited PI3K/AKT signal pathway could reverse the inhibitory effect of CXCL14 on testosterone synthesis and the expression of StAR, CYP11A1 and 3β-HSD in rooster Leydig cells. Our results indicated that melatonin inhibits testosterone synthesis by targeting miR-7481-3p/CXCL14 and inhibiting the PI3K/AKT pathway.

Effect of atrazine on testicular toxicity involves accommodative disorder of xenobiotic metabolizing enzymes system and testosterone synthesis in European quail (Coturnix coturnix)

Due to the wide use of atrazine (ATR), the concern has increased regarding the negative impact of ATR on reproduction. Nevertheless, the reproductive effects caused by different exposure concentrations and the severity of toxic damage are poorly understood. In organisms, ATR is metabolized and degraded through phase II enzyme systems, and changes in cytochrome P450 (CYP) enzymes may have a regulatory role in the harm of ATR. However, less information is available on the induction of CYPs by ATR in avian organisms, and even less on its effects on the testis. Birds are exposed to ATR mainly through food residues and contaminated water, the purpose of this study was to examine reproductive toxicity by different exposure concentrations and elaborate metabolic disorders caused by ATR in European quail (Coturnix coturnix). In this study, the quail were given ATR at 50 mg/kg, 250 mg/kg and 500 mg/kg by oral gavage for 45 days, and the testicular weight coefficients, histopathology and ultrastructure of testes, primary biochemical functions, sex steroid hormones, critical protein levels in the testosterone synthesis pathway, the expression of genes involved CYPs, gonad axis and nuclear receptors expression were investigated. Altogether, testicular coefficient decreased significantly in the high-dose group (1.22%) compared with the control group (3.03%) after 45 days of ATR exposure, and ATR is a potent CYP disruptor that acts through the NXRs and steroid receptor subfamily (APND, CAR, ERND and ERα) without a dose-dependent manner. Notably, ATR interfered with the homeostasis of hormones by triggering the expression of hormones on the gonad axis (LH and E2). These results suggest that exposure to ATR can cause testicular toxicity involving accommodative disorder of phase II enzyme and testosterone synthesis in European quail.

ZNACZENIE RECEPTORA ANDROGENOWEGO W TERAPII RAK STERCZA

he androgen receptor, upon binding to a ligand, functions as a transcription factor, ultimately helping to maintain the proper structure and function of the prostate. In prostate cancer, the constant activation of the androgen pathway in the cell leads to the proliferation of cancer cells and the development of prostate cancer. The individual stages of androgen signaling have become the target of anti-androgen therapy, which ultimately results in a decrease in testosterone levels in the cell. Over time, the duration of therapy leads to the emergence of new mechanisms of resistance in prostate cancer cells, such as an increase in the number of androgen receptors or endogenous synthesis of testosterone. This ultimately leads to the evolution of cancer cells into castration-resistantprostate cancer. This article aims to review and compare contemporary anti-hormonal drugs used in prostate cancer. New generation drugs such as enzalutamide, apalutamide, and darolutamide bind to the androgen receptor and inhibit its translocation to the cell nucleus. Abiraterone acetate inhibits endogenous testosterone synthesis, while relugolix is the first oral gonadotropin-releasing hormone analogue that does not cause a “flare” effect. Understanding the molecular mechanisms and biology of the androgen receptor is crucial for future directions in the development of prostate cancer drug therapies and their appropriate use.

Protective effect of TNFAIP3 on testosterone production in Leydig cells under an aging inflammatory microenvironment

BackgroundThe aging inflammatory microenvironment surrounding Leydig cells is linked to reduced testosterone levels in males. Tumor necrosis factor alpha-induced protein 3 (TNFAIP3) acts as a critical anti-inflammatory factor in various aging-related diseases. This study aims to investigate the protective effect of TNFAIP3 on testosterone production in Leydig cells under an aging inflammatory microenvironment. MethodsBioinformatics analysis examined TNFAIP3 expression differences in aging rat testes and validated the findings in aging mouse testes. In vitro models of inflammation were established using two Leydig cell lines, with tumor necrosis factor alpha (TNF-α) as the inflammatory factor. Lentiviral transduction was utilized to manipulate TNFAIP3 expression in these cell lines. Transcriptomic sequencing identified differentially expressed genes in TNFAIP3-overexpressing cells. ResultsBioinformatics analysis and validation experiments revealed increased inflammatory signaling and elevated TNFAIP3 expression in aging rat and mouse testes. TNFAIP3 knockdown worsened testosterone synthesis inhibition and apoptosis in cells, while TNFAIP3 overexpression reversed these effects. Transcriptome analysis identified alterations in the P38MAPK pathway following TNFAIP3 overexpression. TNFAIP3 knockdown enhanced TNF-induced P38MAPK signaling, whereas its overexpression attenuated this effect. TNFAIP3 was found to regulate testosterone synthesis by upregulating CEBPB expression. ConclusionsTNFAIP3 exhibits inhibitory effects on apoptosis and promotes testosterone production in Leydig cells. The protective influence of TNFAIP3 on Leydig cells within an inflammatory microenvironment is likely mediated through by inhibiting the P38MAPK pathway and upregulating CEBPB expression.

Challenging Stereotypes: Cases of Breast Cancer in Men

Breast cancer is the most common malignant tumor in women, internationally, and the leading cause of death from cancer. However, breast cancer in men is an uncommon pathology, corresponding to less than 1% of neoplasms in men. The lifetime risk of breast cancer in a man is approximately 1:1000, compared to 1:8 for a woman. Currently, an increase in timely diagnosis has been demonstrated thanks to the programs that are carried out for the early detection of breast cancer worldwide. Although, unfortunately, early diagnosis is still difficult due to the lack of information that a large part of the population has regarding self-examination, not only in our country, but also around the world, and in the case of breast cancer in men, being such an uncommon condition, its diagnosis is more difficult. The incidence of breast cancer is increasing in the developing world, due to longer life expectancy, increased urbanization and the adoption of Western lifestyles. Among the risk factors that have been seen to increase the risk of breast cancer in men, with scientifically proven evidence, are mainly hormonal disorders, family history and mutations of certain genes that predispose to the disease. The hormonal disorders mentioned above refer to an increase in circulating estrogen levels and a deficiency in the synthesis or action of testosterone. These alterations can be caused by disorders at the level of testicular testosterone synthesis, such as: orchitis, orchiectomy, undescended testicle, congenital inguinal hernia and others. Patients who voluntarily consume exogenous estrogens as part of the treatment of prostate cancer or transsexuals should also be taken into account. Another risk factor that has been found to be implicated in the presentation of this type of neoplasia are medications; many frequently used medications are testosterone antagonists. Alcohol is also implicated, since it decreases testosterone synthesis and the number of testicular receptors for gonadotropin. A higher prevalence has also been seen in patients with obesity, in whom there is a double risk, because as they have a higher peripheral aromatization of androgens, they synthesize a greater amount of circulating estrogens. Approximately 20% of men with breast cancer have a first-line family history, which indicates a two to three times higher risk of developing the disease at some point in their lives and that this risk doubles exponentially with the number of affected family members. Currently there are many programs and organizations that promote the fight against breast cancer, such as the World Health Organization, where there are comprehensive national cancer control programs that are integrated with non-communicable diseases and other related problems. Comprehensive cancer control encompasses prevention, early detection, diagnosis and treatment, rehabilitation and palliative care. Raising awareness of the general public about the problem of breast cancer and control mechanisms, as well as promoting appropriate policies and programs, are key strategies for population-based breast cancer control. Based on what has been mentioned above, the work will be carried out in order to have a better approach to breast cancer in men, due to the little information we have about it, as well as to raise awareness among the general population about the risk factors, prognostic factors, timely diagnosis and treatment of patients with this type of pathology.

Pyrroloquinoline quinone inhibits PCSK9-NLRP3 mediated pyroptosis of Leydig cells in obese mice

Abnormal lipid metabolism and chronic low-grade inflammation are the main traits of obesity. Especially, the molecular mechanism of concomitant deficiency in steroidogenesis-associated enzymes related to testosterone (T) synthesis of obesity dominated a decline in male fertility is still poorly understood. Here, we found that in vivo, supplementation of pyrroloquinoline quinone (PQQ) efficaciously ameliorated the abnormal lipid metabolism and testicular spermatogenic function from high-fat-diet (HFD)-induced obese mice. Moreover, the transcriptome analysis of the liver and testicular showed that PQQ supplementation not only inhibited the high expression of proprotein convertase subtilisin/Kexin type 9 (PCSK9) but also weakened the NOD-like receptor family pyrin domain containing 3 (NLRP3)-mediated pyroptosis, which both played a negative role in T synthesis of Leydig Cells (LCs). Eventually, the function and the pyroptosis of LCs cultured with palmitic acid in vitro were simultaneously benefited by suppressing the expression of NLRP3 or PCSK9 respectively, as well the parallel effects of PQQ were affirmed. Collectively, our data revealed that PQQ supplementation is a feasible approach to protect T synthesis from PCSK9-NLRP3 crosstalk-induced LCs’ pyroptosis in obese men.

Adropin maypromote insulin stimulated steroidogenesis and spermatogenesis in adult mice testes.

Adropin is a versatile peptide which was discovered as a novel metabolic hormone that is involved in the regulation of lipid and glucose homeostasis. However, its possible role in the testicular function is not yet understood. The aim of our study was to explore the distribution pattern of adropin and GPR19 in various cell types and its possible role in testicular functions of adult mice. Immunohistochemical study revealed the intense immunoreactivity of adropin in the Leydig cells, while GPR19 showed intense immunoreactivity in the pachytene spermatocytes and mild immunoreactivity in Leydig cells and primary as well as secondary spermatocytes in mouse testis. Enho mRNA was also found to be expressed in the mouse testis. These findings suggested that adropin-GPR19 signaling may act in autocrine/paracrine manner to modulate testicular functions. Furthermore, to find out the direct role of adropin in the testicular function, in vitro study was performed in which testicular slices were cultured with adropin alone (10 and 100 ng/mL) and in combination with insulin (5 μg/mL). Adropin alone inhibited testicular testosterone synthesis by inhibiting the expression of P450-SCC, 3β-HSD, and17β-HSD while along with insulin stimulated the testicular testosterone synthesis by increasing the expression of GPR19, IR, StAR, P450-SCC, 3β-HSD, and 17β-HSD. Adropin alone or in combination with insulin promoted germ cell survival and proliferation by upregulating the expression of PCNA, Bcl2, and pERK1/2. Thus, it can be concluded that adropin-GPR19 signaling promotes insulin stimulated steroidogenesis and germ cell survival as well as proliferation in the mice testes in an autocrine/paracrine manner.

Co-exposure to polystyrene microplastics and microcystin-LR aggravated male reproductive toxicity in mice

Microplastics (MPs) are plastic pollutants with a diameter of less than 5 mm and microcystins (MCs) are natural toxins produced by cyanobacteria. In recent years, the pollution of MPs and MCs attracted widespread attention. However, our understanding about the toxic effects of co-exposure of MPs and MCs on male reproduction is limited. Mice were continuously exposed to 0.04mg/(kg*bw) microcystin-leucine-arginine (MC-LR) or 45 mg/(kg*bw) polystyrene microplastics (PS-MPs) or a mixed solution of 0.04mg/(kg*bw) MC-LR and 45 mg/(kg*bw) PS-MPs by gavage for 28 days in this study. The results showed that PS-MPs could absorb MC-LR in ddH2O and MC-LR content in testis was increased in the group with combined exposure when compared to the group only exposed to MC-LR. Exposure to PS-MPs or MC-LR individually could destroy testis structure, increase the level of tissue apoptosis and decrease the quality of sperm, while the co-exposure enhanced the toxic effects. Furthermore, PS-MPs could carry MC-LR into testis Leydig cells, reduce testosterone levels and mRNA expression levels of key molecules involved in testosterone synthesis (StAR, P450scc, P450c17,3β-HSD and 17β-HSD). Among them, the combined effect of PS-MPs-MC-LR was the most severe. In summary, this study provides new insights into the toxicity of MPs and MCs in mammals.

Small Molecule Cocktails Promote Fibroblast-to-Leydig-like Cell Conversion for Hypogonadism Therapy.

Male hypogonadism arises from the inadequate production of testosterone (T) by the testes, primarily due to Leydig cell (LC) dysfunction. Small molecules possess several advantages, including high cell permeability, ease of synthesis, standardization, and low effective concentration. Recent investigations have illuminated the potential of small molecule combinations to facilitate direct lineage reprogramming, removing the need for transgenes by modulating cellular signaling pathways and epigenetic modifications. In this study, we have identified a specific cocktail of small molecules, comprising forskolin, DAPT, purmorphamine, 8-Br-cAMP, 20α-hydroxycholesterol, and SAG, capable of promoting the conversion of fibroblasts into Leydig-like cells (LLCs). These LLCs expressed key genes involved in testosterone synthesis, such as Star, Cyp11a1, and Hsd3b1, and exhibited the ability to secrete testosterone in vitro. Furthermore, they successfully restored serum testosterone levels in testosterone-castrated mice in vivo. The small molecule cocktails also induced alterations in the epigenetic marks, specifically H3K4me3, and enhanced chromosomal accessibility on core steroidogenesis genes. This study presents a reliable methodology for generating Leydig-like seed cells that holds promise as a novel therapeutic approach for hypogonadism.

Yangjing Capsule Increases Testosterone Production through SET/PI3K/Akt Pathway in Leydig Cell

Background. Our previous works revealed Yangjing capsule (YC) could inhibit cell apoptosis and promote testosterone production in Leydig cells. SET protein has been reported to be important in regulating testosterone synthesis and apoptosis. The purpose of this study was to investigate whether the steroidogenic and antiapoptotic effects of YC are mediated in part by the SET/PI3K/Akt pathway in Leydig cells. Methods. MLTC-1 cells were treated with YC-medicated serum. si-RNA was used to knockdown SET expression in MLTC-1 cells. The elderly BALB/c mice were treated with different doses of YC extract. Flow cytometry and TUNEL staining were used to assess apoptosis in MLTC-1 cells and mouse testes. HE staining was conducted to detect the morphological changes in mouse testes. Testosterone levels in cell culture medium and serum were measured by ELISA kit. Quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression levels of key molecules. Results. YC-medicated serum could significantly increase the expression of SET and StAR, P450scc, HSD17B, and testosterone production in MLTC-1 cells. YC-medicated serum could increase Akt phosphorylation and Bcl-2/Bax ratio to suppress apoptosis in MLTC-1 cells. SET knockdown significantly inhibited YC-medicated serum-induced steroidogenic and antiapoptotic effects in MLTC-1 cells. In vivo experiments revealed the YC extract could enhance SET expression, trigger Akt phosphorylation, and suppress apoptosis in mouse testis to raise serum testosterone levels. Conclusions. A possible mechanism of promoting testosterone production induced by YC in Leydig cell was by triggering SET/PI3K/Akt pathway.