Comparative Analysis Among Different Species Reveals That the Androgen Receptor Regulates Chicken Follicle Selection Through Species-Specific Genes Related to Follicle Development

The differences in reproductive processes at the molecular level between viviparous and oviparous animals are evident, and the site in the ovary that synthesizes sex hormones (androgens and oestrogens) and the trends for enriching sex hormones during follicle development in chickens are different from those in mammals, suggesting that the effect of sex hormones on follicle development in chickens is probably different from that in viviparous animals. To explore the specific role of androgen receptors (ARs) on chicken follicular development, we matched the correspondence of follicular development stages among chickens, humans, cows and identified chicken-specific genes related to follicle development (GAL-SPGs) by comparing follicle development-related genes and their biological functions among species (chickens, humans, and cows). A comparison of the core transcription factor regulatory network of granulosa cells (or ovaries) based on super-enhancers among species (chicken, human, and mouse) revealed that AR is a core transcriptional regulator specific to chickens. In vivo experiments showed that inhibition of AR significantly reduced the number of syf (selected stage follicles) in chickens and decreased the expression of GAL-SPGs in F5 follicles, while in vitro experiments showed that inhibition of AR expression in chicken granulosa cells (GCs) significantly down-regulated the expression levels of GAL-SPGs, indicating that AR could regulate follicle selection through chicken-specific genes related to follicle development. A comparison among species (77 vertebrates) of the conserved genomic regions, where chicken super-enhancers are located, revealed that the chicken AR super-enhancer region is conserved in birds, suggesting that the role of AR in follicle selection maybe widespread in birds. In summary, we found that AR can regulate follicle selection through chicken-specific genes related to follicle development, which also emphasizes the important role of AR in follicle selection in chickens and provides a new perspective for understanding the unique process of follicle development in chickens. Our study will contribute to the application of androgens to the control of egg production in chickens and suggests that researchers can delve into the mechanisms of follicle development in birds based on androgen/androgen receptors.

Receptor-related aspects of reduced lubrication in women of reproductive age with hypoandrogenism

Aim — to investigate receptivity of the vaginal epithelium in women with the female sexual dysfunction and hypoandrogenism.Materials and methods. The immunohistochemical investigation has been performed to define the density of androgen receptors (AR) and estrogen receptors (ER) in the distal part of the vaginal tract of women of reproductive age with hypoandrogenic provision of their hormonal background. For immunohistological examination, a biopsy of the lower third of the vaginal mucosa was taken using a dermopunch with a diameter of 3 mm in 20 women with female sexual dysfunction (FSD) and hypo­andro­genism. The control group consisted of 20 women of reproductive age without FSDmanifestations and disturbances in their hormonal levels. The survey was carried out at the stage of pre-conceptional preparation. Location: Ukrainian Scientific and Prac­tical Center for Endocrine Surgery, Transplantation of Endocrine Organs and Tissues of the Ministry of Health of Ukraine.Results. Immunoreactivity of ERα in the epithelium in the main and control groupswas 47.2 ± 22.2 and 86.1 ± 8.0 (p < 0.001), in the stroma — respectively 13.9 ± 16.8 and 86.1 ± 8.3 (p < 0.001), that is, immunoreactivity in the epithelium was significantly higher thanin the stroma (p < 0,01). The ERα locationin the vaginal epithelium was the samein both groups: cellsof the basal-intermediate layer along the basement membrane and the intermediate layer. No cells, expressing ERα, were detected in the surface layer. In stromal cells, ERαcontained in fibroblasts and smooth muscle cells. ERβ was found in cellsof thebasal — parabasal layer, the intermediate layer, occasionally —on the surface layerof vaginal epithelium, as well as in fibroblasts, vascular endothelium, where both nuclear andcytoplasmic staining was revealed. According to the results of immunohistochemical study of AR invagina, the proportion of AR-positive cells in healthy women was negligible (3.8 ± 2.2), that is considerably less than in patients with androgen deficiency (18.8 ± 8.3) and compared withother receptors. AR were localized mainly in the basal layer alongbasement membrane. Single AR-positive fibroblasts occurred in the stroma.Conclusions. Estrogen receptors α (ERα) are involved in the regulation of the processes of proliferation and differentiation of the vaginal epithelium under the influence of estrogen, as evidenced by a 1.8-fold increase in the density of stained ERα in women of the control group. The absence of correlations between ЕRα and ЕRβ receptors (r = 0.24; p < 0.05) may indicate the ability to change in the same tissue depending on the strength of estrogen effects on the body of a woman. Topical localization of androgen receptors in the basal layer testifies in favour of the safe use of local formsof androgensto overcome the reduced lubrication.

Influence of androgen deficiency on the endometrium structure in women of reproductive age

Aim — to identify the relationship between androgen deficiency and the development of endometrial hypoplasia in women of reproductive age, to develop an algorithm for the diagnosis and treatment of this category of women. Materials and methods. Examination of patients with androgen deficiency revealed 48 patients with endometrial hypoplasia based on the ultrasound markers. After examination for CD138 and detection of chronic endometritis during the study, 9 patients were excluded. At the second stage, an immunohistochemical examination was performed for the expression of receptors for estrogen, progesterone and androgens. According to the results, the patients were divided into 2 groups: the first group (24 patients) with a high level of expression of androgen receptors and the second group (15 patients) with a low level of expression of androgen receptors. Theexpressionof receptors to estrogens and progesterone was on medium level and comparative in both groups. Both groups of patients underwent hormonal therapy for 3 months: estradiol valerate 1 g per day in a continuous mode and 200 mg of micronized progesterone from the sixteenth to twenty-fifth days of the menstrual cycle. Additionally, patients of the first group received dehydro­epian­drosterone(DHEA)in a dose of 25 mg per day continuously in the form sublingual spray. Results. According to the data of ultrasound examination in the first group of patients, the endometrium corresponded to normal parameters both during treatment and 1 and 3 months after stopping treatment. At the same time, in the second group of patients, there was an improvement in the thickness (more than 7 mm) and structure of the endometrium during treatment and the absence of these effects after the termination of hormonal therapy. Considering the recommendations of the Association of Endocrinologists on the superiority of non-tablet forms of androgen preparations in the treatment of androgen deficiency and having a positive and long-term effect when taking sublingual DHEA, it is possible to recommend adding the above form of DHEA to systemic therapy of endometrial hypoplasia against the background of androgen deficiency. Conclusions. Women with androgen deficiency are more likely to have concomitant endometrial hypoplasia. Immunohistochemical examination of the endometrium of women of reproductive age with androgen deficiency in 24 patients (61.5 %) revealed a high level of expression of androgen receptors. The effectiveness of therapy for endometrial hypoplasia in women with androgen deficiency with addition of androgens to the standard regimens is more effective and has a long-lasting effect. The combination of estrogen-gestagenic therapy and androgens has a positive effect on the gestational potential of the endometrium in women of reproductive age with androgen deficiency.

Prognostic Significance of Tumor–infiltrating Lymphocytes and Androgen Receptors in Patients with Early Triple-negative Breast Cancer

Aims This study aimed to investigate the association between tumor-infiltrating lymphocytes (TIL) and androgen receptors (AR) and to assess their impact on early triple-negative breast cancer (TNBC) prognosis. Previous studies analyzed only stromal TIL (sTIL) and intratumoral (ITIL), while this study includes an additional spatial analysis for TIL in central tumor (CT) and invasive margin (IM) compartments and correlation with AR expression and overall survival (OS). Methods A retrospective cohort study encompassing 152 early TNBC tissue samples from patients treated at a tertiary oncologic center between 2009 and 2012. TIL and AR were assessed using formalin-fixed paraffin-embedded samples, using hematoxylin-eosin staining and immunohistochemistry. AR-positive tumors were considered those with ≥ 1% nuclear-stained cells. Results High TIL indicators were found to be positive prognostic factors. Although AR was not an independent prognostic factor, its interactions with sTIL and ITIL at IM impacted OS. Positive AR along with high sTIL and ITIL in IM were associated with favorable OS (HR for sTIL 0.22; 95%CI 0.05–0.97; p = 0.045 and HR for ITIL 0.10; 95%CI 0.01–0.78; p = 0.028). Conclusion Spatial morphological analysis of TIL reveals an additional prognostic value when combined with AR status, and shows a clinically significant impact on OS in early TNBC.

The Prostate Saturation Point after Testosterone Replacement Therapy in Testosterone Deficiency Patient

Background: The effect of testosterone on the prostate gland is an unresolved question. The prostate saturation model is a recent hypothesis explaining that the stimulation of prostate tissue by testosterone is limited to a certain level of testosterone due to the limited number of androgen receptors. However, data from the Thai patients related to this issue are still lacking and need to be explored. Objective: To investigate prostate changes after testosterone replacement therapy (TRT). Materials and Methods: A retrospective study including testosterone-deficient patients who had TRT between 2011 and 2017 at Ramathibodi Hospital was conducted. The change in prostate-specific antigen (PSA) levels before and after TRT, or after a 1-year observation, was measured and analyzed as the primary objective. As a secondary objective, the authors measured and evaluated normal PSA velocity (PSAV) in the patients after TRT. Results: One hundred eleven testosterone deficient patients were included for analysis. The mean age was 62 years old. The baseline testosterone level and PSA level at the beginning were 247 ng/dL and 1.16 ng/mL, respectively. After undergoing TRT for one year, the results showed that the testosterone and the PSA levels were 307 ng/dL and 1.46 ng/mL, respectively. In addition, the subgroup analysis illustrated that patients who had low baseline testosterone levels such as 247 ng/dL or less, had significant increase of PSA level after treatment. However, when the baseline testosterone level was more than 247 ng/dL, the PSA levels were steady after treatment. For the secondary-objective results, the PSAV of the testosterone deficiency patients after TRT was 0.3 ng/mL/year. Conclusion: The evidence clearly indicates that TRT significantly increased the serum testosterone level. However, it had a limited effect on PSA change. The present study results supported the hypothesis of the prostate saturation model. The authors believe that a testosterone level of 247 ng/dL can saturate all androgen receptors in the prostate gland and no longer increase prostate stimulation. Keywords: Prostate-specific antigen; Prostate cancer; Testosterone replacement Therapy; Prostate saturation

Nobiletin Protects Against Diabetes-induced Testicular Injury via Hypophysis-gonadal Axis Up-regulation and Amelioration of Oxidative Stress

Background: Testicular injury is one of the most serious problems of Diabetes mellitus. The present study aims to compare the effect of two different doses of nobiletin and the probable mechanisms against diabetes-induced testicular impairment in rats. Methods and Results: Streptozotocin injection was used to induce diabetes. Diabetic rats received nobiletin (10 mg/kg) or (25 mg/kg) daily and orally for 30 days. Diabetic rats displayed a significant elevation in glucose, glycosylated hemoglobin (HbA1c), homeostasis model of insulin resistance (HOMA-IR), and pro-inflammatory cytokines. Levels of serum insulin, testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were significantly reduced. Histological changes with positive caspase-3 and decreased Androgen receptors (AR) immunoexpressions were observed in diabetic rats. Both doses of nobiletin improved hyperglycemia, reduced pro-inflammatory cytokines, and augmented insulin, testosterone, LH, and FSH levels. Gene and protein manifestation of LH and FSH receptors and cytochrome P450 17 α-hydroxylase (CYP17A1) was markedly down-regulated in testicular tissues of diabetic group, an effect which was markedly increased with both doses of nobiletin. In addition, both doses significantly reduced lipid peroxidation, and caspase-3 immuno-expression and improved the activity of the antioxidant enzymes and AR in testicular tissues of diabetic group. Conclusion: Both doses showed protective effects against diabetes-induced testicular injury by diminution of oxidative stress, hyperglycemia, inflammation, caspase-3 and up-regulation of the hypophysis-gonadal axis and androgen receptors. The high dose of nobiletin was more effective than the lower dose.