RESUMO
INTRODUCTION: Hair loss is driven by multiple factors, including genetics. Androgenetic alopecia (AGA) is a condition in which treatments necessitate prolonged compliance with prescribed medications. We have developed IVL3001, a long-acting injectable (LAI) formulation of finasteride encapsulated within poly lactic-co-glycolic acid microspheres, to enhance the efficacy of the finasteride and to achieve consistent positive outcomes in adults. An open-label, sequential, single-dose phase I clinical trial was designed to evaluate the safety, pharmacokinetic (PK), and pharmacodynamic (PD) of IVL3001. METHODS: A total of 40 non-smoking, healthy adult males were divided into three cohorts where the IVL3001 group received a single subcutaneous injection of 12-36 mg IVL3001 and 1 mg finasteride (Propecia®) once daily for 28 days. The plasma concentrations of finasteride, dihydrotestosterone (DHT), and testosterone were measured using liquid chromatography-tandem mass spectrometry. The tolerability of the injections was assessed, and compartment models were developed to predict the effective dose and assess PK/PD profiles. RESULTS: IVL3001 and finasteride 1 mg tablets were well tolerated. IVL3001 showed consistent plasma concentrations without bursts or fluctuations. Consistent with its mechanism of action, IVL3001 reduced DHT levels. Simulation data showed that the administration of 12-36 mg of IVL3001 every 4 weeks achieved plasma concentrations similar to finasteride, with comparable DHT reduction. CONCLUSION: The present study represents the first clinical trial to evaluate the safety, pharmacokinetic (PK), pharmacodynamic (PD), and tolerability of finasteride long-acting injectables (LAI) in adults. The rapid onset of action sustained effective drug concentration and the prolonged half-life of IVL3001 suggest that it offers multiple benefits over conventional oral formulations in terms of therapeutic response and compliance. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT04945226.
Assuntos
Inibidores de 5-alfa Redutase , Alopecia , Finasterida , Humanos , Finasterida/farmacocinética , Finasterida/administração & dosagem , Finasterida/efeitos adversos , Alopecia/tratamento farmacológico , Masculino , Adulto , Inibidores de 5-alfa Redutase/farmacocinética , Inibidores de 5-alfa Redutase/administração & dosagem , Inibidores de 5-alfa Redutase/efeitos adversos , Inibidores de 5-alfa Redutase/farmacologia , Di-Hidrotestosterona/farmacocinética , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/sangue , Pessoa de Meia-Idade , Preparações de Ação Retardada , Testosterona/farmacocinética , Testosterona/sangue , Injeções Subcutâneas , Adulto Jovem , MicroesferasRESUMO
Hyperandrogenemia (HA) is a hallmark of polycystic ovary syndrome (PCOS) and is an integral element of non-alcoholic fatty liver disease (NALFD) in females. Administering low-dose dihydrotestosterone (DHT) induced a normal weight PCOS-like female mouse model displaying NAFLD. The molecular mechanism of HA-induced NAFLD has not been fully determined. We hypothesized that DHT would regulate hepatic lipid metabolism via increased SREBP1 expression leading to NAFLD. We extracted liver from control and low-dose DHT female mice; and performed histological and biochemical lipid profiles, Western blot, immunoprecipitation, chromatin immunoprecipitation, and real-time quantitative PCR analyses. DHT lowered the 65 kD form of cytosolic SREBP1 in the liver compared to controls. However, DHT did not alter the levels of SREBP2 in the liver. DHT mice displayed increased SCAP protein expression and SCAP-SREBP1 binding compared to controls. DHT mice exhibited increased AR binding to intron-8 of SCAP leading to increased SCAP mRNA compared to controls. FAS mRNA and protein expression was increased in the liver of DHT mice compared to controls. p-ACC levels were unaltered in the liver. Other lipid metabolism pathways were examined in the liver, but no changes were observed. Our findings support evidence that DHT increased de novo lipogenic proteins resulting in increased hepatic lipid content via regulation of SREBP1 in the liver. We show that in the presence of DHT, the SCAP-SREBP1 interaction was elevated leading to increased nuclear SREBP1 resulting in increased de novo lipogenesis. We propose that the mechanism of action may be increased AR binding to an ARE in SCAP intron-8.
Assuntos
Di-Hidrotestosterona/administração & dosagem , Regulação da Expressão Gênica/efeitos dos fármacos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas de Membrana/genética , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Síndrome do Ovário Policístico/metabolismo , Animais , Composição Corporal , Peso Corporal , Feminino , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/metabolismo , Obesidade/induzido quimicamente , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/fisiologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Background Human 3ß-hydroxysteroid dehydrogenase type 1 (HSD3B1) is an enzyme associated with steroidogenesis, however its' role in hepatocellular carcinoma (HCC) biology is unknown. Trilostane is an inhibitor of HSD3B1 and has been tested as a treatment for patients with breast cancer but has not been studied in patients with HCC. Methods and Results The expression of HSD3B1 in HCC tumors in 57 patients were examined. A total of 44 out of 57 tumors (77.2%) showed increased HSD3B1 expression. The increased HSD3B1 in tumors was significantly associated with advanced HCC. In vitro, the knockdown of HSD3B1 expression in Mahlavu HCC cells by a short hairpin RNA (shRNA) led to significant decreases in colony formation and cell migration. The suppression of clonogenicity in the HSD3B1-knockdown HCC cells was reversed by testosterone and 17ß-estradiol. Trilostane-mediated inhibition of HSD3B1 in different HCC cells also caused significant inhibition of clonogenicity and cell migration. In subcutaneous HCC Mahlavu xenografts, trilostane (30 or 60 mg/kg, intraperitoneal injection) significantly inhibited tumor growth in a dose-dependent manner. Furthermore, the combination of trilostane and sorafenib significantly enhanced the inhibition of clonogenicity and xenograft growth, surpassing the effects of each drug used alone, with no documented additional toxicity to animals. HSD3B1 blockade was found to suppress the phosphorylation of extracellular signal-regulated kinase (ERK). The decreased ERK phosphorylation was reversed by testosterone or 17b-estradiol. Conclusions Trilostane significantly inhibited the growth of HCC by inhibiting HSD3B1 function and augmenting the efficacy of sorafenib.
Assuntos
Carcinoma Hepatocelular/patologia , Di-Hidrotestosterona/análogos & derivados , Neoplasias Hepáticas/patologia , Complexos Multienzimáticos/antagonistas & inibidores , Progesterona Redutase/antagonistas & inibidores , Sorafenibe/farmacologia , Esteroide Isomerases/antagonistas & inibidores , Idoso , Animais , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/farmacologia , Quimioterapia Combinada , Estradiol/farmacologia , Feminino , Humanos , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , RNA Interferente Pequeno/efeitos dos fármacos , Sorafenibe/administração & dosagem , Testosterona/farmacologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: The explicit mechanism of erectile dysfunction caused by low androgen status is unknown. It was reported that eNOS was expressed in extracellular vesicles (EVs). Androgen may regulate erectile function by affect the release of EVs from endothelial cells. OBJECTIVES: To investigate whether androgen affects the production of EVs and nitric oxide (NO) in endothelial cells of rat penile corpus cavernosum. MATERIALS AND METHODS: Endothelial cells of rat penile corpus cavernosum were isolated and purified from 6-week-old healthy male Sprague Dawley (SD) rats. Endothelial cells were treated with different concentrations of dihydrotestosterone (DHT) in a cell culture medium as follows: no-androgen group (NA group, DHT 0 nmol/L), very-low androgen group (VLA group, DHT 0.1 nmol/L), low androgen group (LA group, DHT 1 nmol/L), and physiological concentrations androgen group (PA group, DHT 10 nmol/L). After 24 h, EVs of supernatant in each group were isolated and identified. The content of EVs and NO in the supernatant and the expression of CD9, CD63, TSG101, and eNOS in EVs were detected. RESULTS: Positive expression of CD9, CD63, TSG101, and eNOS was found in isolated EVs. The concentration of EVs was lower in the NA group compared with other groups (p < 0.01). The expression of eNOS and the concentration of NO was lower in the NA group than that in other groups (p < 0.05); it was lower in the VLA group than that in the LA group (p < 0.05) and lower in LA group than that in PA group (p < 0.05). When the concentration of DHT in endothelial cell culture medium ranged from 0 to 10 nmol/L, the concentration of DHT was positively correlated with the content of EVs and NO. CONCLUSION: Decrease in eNOS-expressing EVs is one mechanism of NO reduction in endothelial cells of rat corpus cavernosum caused by low androgen levels.
Assuntos
Androgênios/administração & dosagem , Di-Hidrotestosterona/administração & dosagem , Células Endoteliais/efeitos dos fármacos , Vesículas Extracelulares/efeitos dos fármacos , Pênis/efeitos dos fármacos , Animais , Avaliação Pré-Clínica de Medicamentos , Células Endoteliais/metabolismo , Disfunção Erétil/tratamento farmacológico , Vesículas Extracelulares/metabolismo , Masculino , Pênis/metabolismo , Cultura Primária de Células , Ratos Sprague-DawleyRESUMO
We provide the details of the successful management of a patient with active Cushing's disease complicated with coronavirus disease 2019 (COVID-19) pneumonia. The patient was a 27-year-old Japanese female healthcare worker who was scheduled to undergo pituitary surgery for Cushing's disease. She had been in close contact with an undiagnosed patient infected with COVID-19 and then developed COVID-19 pneumonia. Despite a lack of known risk factors associated with severe COVID-19 infection, the patient's dyspnea worsened and her respiratory condition deteriorated, as indicated by the need for 7 L/min oxygen supply by mask to maintain her oxygen saturation at >90%. Medical treatment was initiated to control hypercortisolism by the 'block and replace' regimen using steroidogenesis inhibitors and hydrocortisone. The COVID-19 pneumonia improved with multi-modal treatment including antiviral therapy. One month later, after a negative severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) test result and with appropriate protection against virus transmission to medical staff in the operating room and daily medical care nurses, trans-sphenoidal surgery was performed by our highly experienced pituitary surgeon. One month after the surgery, the patient's basal ACTH and cortisol levels and urinary free cortisol were all under the detection limit. Surgical remission was expected. Since hypercortisolism due to active Cushing's disease may worsen a COVID-19 infection, multi-disciplinary management that includes appropriate and prompt treatment strategies is mandatory in such cases.
Assuntos
Amidas/administração & dosagem , Benzamidinas/administração & dosagem , COVID-19/terapia , Guanidinas/administração & dosagem , Metirapona/administração & dosagem , Hipersecreção Hipofisária de ACTH/terapia , Pregnenodionas/administração & dosagem , Pirazinas/administração & dosagem , Adenoma Hipofisário Secretor de ACT/complicações , Adenoma Hipofisário Secretor de ACT/tratamento farmacológico , Adenoma/complicações , Adenoma/tratamento farmacológico , Adulto , COVID-19/complicações , COVID-19/patologia , Terapia Combinada , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/análogos & derivados , Progressão da Doença , Feminino , Pessoal de Saúde , Heparina/administração & dosagem , Humanos , Japão , Procedimentos Neurocirúrgicos , Hipersecreção Hipofisária de ACTH/sangue , Hipersecreção Hipofisária de ACTH/complicações , Hipersecreção Hipofisária de ACTH/patologia , SARS-CoV-2/fisiologia , Resultado do Tratamento , Combinação Trimetoprima e Sulfametoxazol/administração & dosagemRESUMO
Androgens have been known to inhibit cutaneous wound healing in men and male mice. However, in children with major burn injuries, a synthetic androgen was reported clinically to improve wound healing. The aim of this study is to investigate the role of dihydrotestosterone (DHT) as a new therapeutic approach in treating major burn injury. In the present study, mice received systemic androgen treatment post major burn injury. Wound healing rate and body weight were monitored over 21 days. The serum level of inflammatory cytokines/chemokines were measured using multiplex immunoassays. In addition, splenocyte enumeration was performed by flow cytometry. Healing phases of inflammation, re-epithelialization, cell proliferation and collagen deposition were also examined. In results, DHT treated mice lost less weight and displayed accelerated wound healing but has no impact on hypermetabolism. Mice, after burn injury, displayed acute systemic inflammatory responses over 21 days. DHT treatment shortened the systemic inflammatory response with reduced splenic weight and monocyte numbers on day 14 and 21. DHT treatment also reduced wound infiltrating macrophage numbers. In conclusion, DHT treatment facilitates local wound healing by accelerating the resolution of inflammation, but not through alterations of post-burn hypermetabolic response.
Assuntos
Androgênios/administração & dosagem , Queimaduras/tratamento farmacológico , Di-Hidrotestosterona/administração & dosagem , Cicatrização/efeitos dos fármacos , Androgênios/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Queimaduras/sangue , Queimaduras/imunologia , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Citocinas/sangue , Di-Hidrotestosterona/farmacologia , Modelos Animais de Doenças , Masculino , Camundongos , Baço/efeitos dos fármacos , Baço/imunologiaRESUMO
RATIONALE: Circumcision like any other surgical procedure is not devoid of complications. Serious complications are rare and include iatrogenic hypospadias, glans ischemia/necrosis, and glans amputation, all of which require an emergent treatment. PATIENT CONCERNS: We report here a case of 6 months-old-boy with a superficial glans ischemia following circumcision. DIAGNOSIS: Physical examination revealed a severely cyanotic glans with the moderate edema of the dorsal penile skin. Plasma levels of D-dimer were 8.57âmg/L. Urine passage was unremarkable while color Doppler ultrasonography revealed a normal blood flow. INTERVENTIONS: The patient was successfully treated with subcutaneous injection of enoxaparin (low-molecular-weight heparin) and topical 2.5% dihydrotestosterone. OUTCOMES: The appearance of the glans penis on the 5th day was close to normal while the control levels of D-dimer dropped to the reference range. The patient was discharged from the hospital on the 6th day. At 6-month follow-up, the appearance of the glans penis was normal. LESSONS: Acute glans penis ischemia following circumcision is a rare complication. Its successful treatment with enoxaparin and topical dihydrotestosterone has not been previously reported in the literature.
Assuntos
Circuncisão Masculina/efeitos adversos , Di-Hidrotestosterona/uso terapêutico , Heparina de Baixo Peso Molecular/uso terapêutico , Isquemia/etiologia , Administração Tópica , Di-Hidrotestosterona/administração & dosagem , Humanos , Lactente , Isquemia/tratamento farmacológico , Masculino , Pênis/irrigação sanguíneaRESUMO
BACKGROUND: Results of ACTH stimulation test (ACTHst), pre- and post-trilostane serum cortisol concentrations (SCCs), urine concentration (urine-specific gravity [USG]), and urine cortisol : creatinine ratios (UCCRs) are common variables used to monitor trilostane treatment of dogs with pituitary-dependent hyperadrenocorticism (PDH). However, none has consistently discriminated dogs receiving an adequate dose (A) from those overdosed (O) or underdosed (U). OBJECTIVES: To assess and compare recommended monitoring variables, including serial SCCs in a cohort of dogs with PDH treated with trilostane. ANIMALS: Privately owned dogs with PDH (n = 22) and 3 healthy dogs (controls). METHODS: Prospective, multicenter, 2-day study. On day "a" (randomized): ACTHst was completed. Day "b" (>2 to <7 days later): SCCs were assessed -0.5 hours, immediately before, and 1, 2, 2.5, 3, 3.5, 4, 6, 8, and 12 hours after trilostane administration. On the first study day, urine collected at home was assessed for USG, UCCR and owner opinions regarding PDH were categorized as: A (clinical signs resolved), U (remains symptomatic), or ill (possible O). RESULTS: At 27 pairs of evaluations, 7 dogs were categorized as A, 19 U, and 1 possible O (excluded from the study). There was overlap in SCC results from the A and U dogs at every time point. Results of USG, UCCR, and ACTHst did not discriminate A from U dogs. Trilostane suppresses SCC within 1 hour of administration and its duration of action in most PDH dogs is <8 hours. CONCLUSIONS AND CLINICAL IMPORTANCE: No single variable or group of variables reliably discriminated A dogs from U dogs during trilostane treatment for PDH.
Assuntos
Di-Hidrotestosterona/análogos & derivados , Doenças do Cão/tratamento farmacológico , Inibidores Enzimáticos/uso terapêutico , Hipersecreção Hipofisária de ACTH/veterinária , Hormônio Adrenocorticotrópico/administração & dosagem , Hormônio Adrenocorticotrópico/farmacologia , Animais , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/uso terapêutico , Doenças do Cão/diagnóstico , Cães , Inibidores Enzimáticos/administração & dosagem , Feminino , Hidrocortisona/sangue , Hidrocortisona/urina , Masculino , Hipersecreção Hipofisária de ACTH/diagnóstico , Hipersecreção Hipofisária de ACTH/tratamento farmacológico , Estudos Prospectivos , Gravidade Específica , Urina/químicaRESUMO
PURPOSE: The aim of this prospective study was to determine whether there is a beneficial role of combining gonadotropin administration with testosterone downregulation in non-obstructive azoospermia patients prior to a second time microsurgical testicular sperm extraction after a negative one. METHODS: A total of 40 non-obstructive azoospermia men were recruited from a specialized IVF center from 2014 to 2016. Participants were divided equally into two groups: Group A was subjected to testosterone downregulation alone for 1 month and then combined with gonadotropin administration for 3 months prior to second time testicular sperm extraction; Group B (controls) underwent second time microsurgical testicular sperm extraction without prior hormonal therapy. RESULTS: Mean baseline follicle-stimulating hormone levels of the controls and the cases were 26.9 ± 11.8 and 25.4 ± 8.7, respectively. One month after testosterone downregulation, follicle-stimulating hormone level of the cases was normalized and became 2.4 ± 1.2. There was no statistically significant difference between baseline follicle-stimulating hormone levels of the controls and cases (p = 0.946). Remarkably, two cases were positive after downregulation (10%) and no controls were positive at second testicular sperm extraction (0%). There was no statistically significant difference between sperm retrieval after the second microsurgical testicular sperm extraction in the controls and the cases (p = 0.072). CONCLUSION: Patients who underwent first time testicular sperm extraction with unfavorable outcome due to different techniques may benefit from testosterone downregulation combined with neoadjuvant gonadotropin administration as it had shown positive sperms retrieval in 2 out of the 20 cases, especially those with hypergonadotropic azoospermia.
Assuntos
Azoospermia , Gonadotropina Coriônica/administração & dosagem , Di-Hidrotestosterona/análogos & derivados , Substâncias para o Controle da Reprodução/administração & dosagem , Recuperação Espermática , Adulto , Azoospermia/tratamento farmacológico , Estudos de Casos e Controles , Di-Hidrotestosterona/administração & dosagem , Regulação para Baixo , Humanos , Masculino , Microcirurgia , Terapia Neoadjuvante , Estudos Prospectivos , Testosterona/fisiologiaRESUMO
It is still unclear whether Testosterone (T) increases sexual desire through a stimulation of the androgen receptor in relevant brain regions or through its conversion to estrogens. The aim of this study was to clarify the mechanisms of T facilitation of female sexual desire by assessing the effect of a non-aromatizable androgen (Dihydrotestosterone, DHT) in a validated animal model. Ovariectomized (OVX) Long-Evans rats were treated with oil (O) + O, 10 mcg Estradiol Benzoate (EB) + O, 10 mcg EB + 500 mcg Progesterone (P), O + 500 mcg DHT or 10 mcg EB + 500 mcg DHT (n = 12 per group). EB was administered 48 h, while P and DHT 4 h, prior to 4 sexual behavioral testing sessions in bisected unilevel pacing chambers. Appetitive behaviors (the frequencies of hops/darts and solicitations) were considered as the main outcome measure. Sexual receptivity indexes [lordosis magnitude, expressed as lordosis rating (LR), and lordosis quotient (LQ)], rejection responses, as well as mounts, intromissions and ejaculations received from the male were also coded. The probability of transition among sexual behaviors was evaluated by Transition Matrices; T-Pattern analysis was performed to detect hidden repeated temporal behavioral sequences. Preliminary analyses found no statistically significant differences between the O + O and EB + O groups, therefore we excluded the EB + O group from further analyses. Rats treated with EB + DHT displayed significantly more appetitive behaviors compared to negative controls (O + O and O + DHT), whereas no difference was observed between EB + DHT rats and positive controls (EB + P); noteworthy, a higher number of appetitive behaviors was observed in the O + DHT group compared to the O + O group. Furthermore, rats treated with EB + DHT showed significantly higher receptivity measures (LR and LQ) and received more mounts, intromissions and ejaculations compared to negative controls (O + O and O + DHT), to levels equivalent to EB + P. No differences were detected in female-male mounts or rejection responses among the 4 groups. Under a qualitative perspective, full solicitation was found exclusively in T-patterns of the EB + DHT group, which was also the only one to display T-patterns of higher order encompassing appetitive behaviors-only events. In conclusion, the administration of DHT in EB-primed OVX Long-Evans rats enhances sexual behavior measures. Specifically, DHT seems to stimulate sequences of appetitive behaviors separated from copulative/reproductive measures. Our data support an independent role of androgens in the facilitation of female sexual desire.
Assuntos
Androgênios/administração & dosagem , Di-Hidrotestosterona/administração & dosagem , Estradiol/análogos & derivados , Estrogênios/administração & dosagem , Ovariectomia , Comportamento Sexual Animal/efeitos dos fármacos , Animais , Estradiol/administração & dosagem , Feminino , Progesterona/administração & dosagem , Progestinas/administração & dosagem , Ratos , Ratos Long-EvansRESUMO
B lymphoma Mo-MLV insertion region 1 (BMI1) has been reported to be an oncoprotein. BMI1 represses tumor suppressors to promote cell proliferation, epithelial-mesenchymal transition (EMT), and cancer progression. Although it is known that the expression of BMI1 is increased in many cancer types, the mechanism of BMI1 upregulation is not yet clear. We performed integrative analysis for 3 sets of prostate cancer (PCa) genomic data, and found that BMI1 and androgen receptor (AR) were positively correlated, suggesting that AR might regulate BMI1. Next, we showed that dihydrotestosterone (DHT) upregulated both mRNA and protein levels of BMI1 and that BMI1 was increased in castration-resistant prostate cancer (CRPC) from both human patients and a mouse xenograph model. We further identified an AR binding site in the promoter/enhancer region of BMI1, and confirmed BMI1 as the direct target of AR using gene-editing technology. We also demonstrated that high expression of BMI1 is critical for the development of castration-resistance. Our data also suggest that BMI1-specific inhibitors could be an effective treatment of CRPC.
Assuntos
Complexo Repressor Polycomb 1/genética , Neoplasias de Próstata Resistentes à Castração/genética , Receptores Androgênicos/genética , Animais , Sítios de Ligação/genética , Proliferação de Células/efeitos dos fármacos , Proteínas de Ligação a DNA/genética , Di-Hidrotestosterona/administração & dosagem , Elementos Facilitadores Genéticos/genética , Transição Epitelial-Mesenquimal/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Camundongos , Regiões Promotoras Genéticas/genética , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/patologia , RNA Mensageiro/genética , Análise Serial de Tecidos , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Approaches to downregulate ovarian function in the sexually mature bitch by applying slow release GnRH agonist implants are hampered by the initial stimulation of folliculogenesis (flare up) and the resulting side effects. The present pilot study was designed to test to what extent these effects can be suppressed by simultaneous treatment with the 3ß-hydroxysteroid-dehydrogenase (HSD3B) blocker trilostane (T). Treatment with T in 6-h intervals completely blocked adrenal cortisol production. However, in parallel and concomitant with the increase of LH, progesterone and estradiol levels increased, ending up in pro-estric steroid levels in two of the three dogs. Hormonal changes were reflected in the respective clinical symptoms. During the whole observation period the course of LH concentrations did not indicate downregulation of pituitary function as a result of treatment with the GnRH-agonist Suprelorin®, 4.7â¯mg. The incomplete inhibitory effect of T on the follicular production of sex steroids could be explained by an insufficient transfer of T into the follicular compartment or the existence of a HSD3B isoform in the dog ovary different from the adrenal enzyme. Concerning the lack of downregulation and when accounting for published data different pharmacodynamics/pharmacokinetic activities of GnRH-agonists should be taken into account.
Assuntos
3-Hidroxiesteroide Desidrogenases/antagonistas & inibidores , Di-Hidrotestosterona/análogos & derivados , Cães , Hormônio Liberador de Gonadotropina/agonistas , Ovário/fisiologia , Pamoato de Triptorrelina/análogos & derivados , Animais , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/farmacologia , Regulação para Baixo , Implantes de Medicamento , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/farmacologia , Estradiol , Ciclo Estral/efeitos dos fármacos , Feminino , Hidrocortisona/sangue , Hormônio Luteinizante , Progesterona/sangue , Pamoato de Triptorrelina/administração & dosagem , Pamoato de Triptorrelina/farmacologiaRESUMO
In previous studies, carried out in humans, we showed that females are resistant to Leishmania mexicana infection. We also showed that 17ß-estradiol (E2) induces killing of parasites inside of murine macrophages. In this work, we compared, for the first time, L mexicana survival inside of male (male BMDM) and female (female BMDM) bone marrow-derived macrophages (BMDM) treated in vitro with E2 or dihydrotestosterone (DHT). We also compared their levels of nitric oxide (NO), interleukin (IL)-6, IL-10, IL-12p70 and tumour necrosis factor (TNF-α). We found that female BMDM are a lot less susceptible to infection as compared with male BMDM. 17ß-estradiol induced killing of most parasites inside of female BMDM. Dihydrotestosterone, on the other hand, induced some parasite killing inside of some infected male BMDM. Interleukin-6 levels were higher in female BMDM treated with either hormone. Neither TNF-α nor IL-10 levels showed significant differences compared with sham controls. Interestingly IL-12p70 was more abundantly produced by sham female BMDM as compared with sham male BMDM. Only female BMDM treated with E2 trigger a robust IL-12p70 production, but it was significantly reduced in male BMDM. This suggests IL-12p70 is an important factor in female-macrophage resistance to L mexicana parasites.
Assuntos
Estradiol/metabolismo , Interleucina-12/imunologia , Leishmania mexicana/fisiologia , Leishmaniose Cutânea/imunologia , Macrófagos/imunologia , Animais , Citocinas/análise , Di-Hidrotestosterona/administração & dosagem , Estradiol/administração & dosagem , Feminino , Humanos , Leishmaniose Cutânea/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Óxido Nítrico/análise , Fatores SexuaisRESUMO
Many women with hyperandrogenemia suffer from irregular menses and infertility. However, it is unknown whether androgens directly affect reproduction. Since animal models of hyperandrogenemia-induced infertility are associated with obesity, which may impact reproductive function, we have created a lean mouse model of elevated androgen using implantation of low dose dihydrotestosterone (DHT) pellets to separate the effects of elevated androgen from obesity. The hypothalamic-pituitary-gonadal axis controls reproduction. While we have demonstrated that androgen impairs ovarian function, androgen could also disrupt neuroendocrine function at the level of brain and/or pituitary to cause infertility. To understand how elevated androgens might act on pituitary gonadotropes to influence reproductive function, female mice with disruption of the androgen receptor (Ar) gene specifically in pituitary gonadotropes (PitARKO) were produced. DHT treated control mice with intact pituitary Ar (Con-DHT) exhibit disrupted estrous cyclicity and fertility with reduced pituitary responsiveness to GnRH at the level of both calcium signaling and LH secretion. These effects were ameliorated in DHT treated PitARKO mice. Calcium signaling controls GnRH regulation of LH vesicle exotocysis. Our data implicated upregulation of GEM (a voltage-dependent calcium channel inhibitor) in the pituitary as a potential mechanism for androgen's pathological effects. These results demonstrate that gonadotrope AR, as an extra-ovarian regulator, plays an important role in reproductive pathophysiology.
Assuntos
Di-Hidrotestosterona/metabolismo , Gonadotrofos/metabolismo , Hiperandrogenismo/complicações , Infertilidade/metabolismo , Receptores Androgênicos/metabolismo , Animais , Sinalização do Cálcio/fisiologia , Células Cultivadas , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/sangue , Modelos Animais de Doenças , Implantes de Medicamento/administração & dosagem , Ciclo Estral/fisiologia , Feminino , Hormônio Liberador de Gonadotropina/metabolismo , Humanos , Hiperandrogenismo/sangue , Hiperandrogenismo/metabolismo , Hiperandrogenismo/fisiopatologia , Sistema Hipotálamo-Hipofisário , Infertilidade/sangue , Infertilidade/patologia , Infertilidade/fisiopatologia , Hormônio Luteinizante/metabolismo , Camundongos , Camundongos Transgênicos , Proteínas Monoméricas de Ligação ao GTP/metabolismo , Ovário/metabolismo , Ovário/patologia , Cultura Primária de Células , Receptores Androgênicos/genética , Regulação para CimaRESUMO
The spinal cord is essential for neuronal communication between the brain and rest of the body. To gain further insight into the molecular changes underpinning maturation of the mouse spinal cord, we analysed gene expression differences between 4 weeks of age (prior to puberty onset) and adulthood (8 weeks). We found 800 genes were significantly differentially expressed between juvenile and adult spinal cords. Gene ontology analysis revealed an overrepresentation of genes with roles in myelination and signal transduction among others. The expression of a further 19 genes was sexually dimorphic; these included both autosomal and sex-linked genes. Given the presence of steroid hormone receptors in the spinal cord, we also looked at the impact of two major steroid hormones, oestradiol and dihydrotestosterone (DHT) on spinal cord gene expression for selected genes. In gonadectomised male animals, implants with oestradiol and DHT produced significant changes to spinal cord gene expression. This study provides an overview of the global gene expression changes that occur as the spinal cord matures, over a key period of maturation. This confirms that both age and sex are important considerations in studies involving the spinal cord.
Assuntos
Envelhecimento/metabolismo , Regulação da Expressão Gênica , Proteínas do Tecido Nervoso/genética , Caracteres Sexuais , Medula Espinal/metabolismo , Envelhecimento/genética , Animais , Castração , Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/farmacologia , Implantes de Medicamento , Estradiol/administração & dosagem , Estradiol/farmacologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Ontologia Genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Bainha de Mielina/fisiologia , Proteínas do Tecido Nervoso/biossíntese , RNA Mensageiro/biossíntese , Maturidade Sexual , Transdução de Sinais/genética , Medula Espinal/crescimento & desenvolvimento , Transcrição GênicaRESUMO
Steroid 5α-reductase type 2 deficiency (5αRD2) is a congenital disorder of sex development caused by impairment of conversion from testosterone (T) to 5α-dihydrotestosterone (DHT). DHT deficiency leads to various degrees of undervirilized external genitalia including micropenis, primarily correlated with mutations of the SRD5A2 gene that encodes 5α-reductase type 2. Four Japanese boys with isolated micropenis were diagnosed as 5αRD2 by elevated ratios of serum T/DHT, and decreased ratios of urinary 5α/5ß-reduced steroid metabolites. Genetic analyses for SRD5A2 identified that the four patients shared a hypomorphic mutation R227Q that has a residual activity related to the mild-form of 5αRD2. For prepubertal micropenis, DHT was transdermally applied to the four patients at the ages of 4-11 year, increasing a median of stretched penile lengths (SPLs) from 2.6 cm (-2.5 SD) to 4.4 cm (-0.2 SD). Nevertheless, the post-pubertal penile growth was apparently retarded, despite normal levels of T secreted from well-developed testes. The second course of DHT treatment underwent at ages of 12-18 year, but unable to normalize SPLs at a range of 6.0 to 7.0 cm (-3.4 to -2.4 SD). The prostate volumes of two patients were variable at 8.1 and 21 cm3, and a sperm cell count of one patient was normal as young adult. DHT treatment contributes to development of the penis and prostate, which are favorable for the potential fertility of 5αRD2 adults. Meanwhile, the retarded penile growth and a risk of prostate overgrowth may complicate the post-pubertal management with DHT for 5αRD2 males.
Assuntos
3-Oxo-5-alfa-Esteroide 4-Desidrogenase/deficiência , Di-Hidrotestosterona/administração & dosagem , Transtorno 46,XY do Desenvolvimento Sexual/tratamento farmacológico , Doenças dos Genitais Masculinos/tratamento farmacológico , Hipospadia/tratamento farmacológico , Pênis/anormalidades , Pênis/efeitos dos fármacos , Puberdade/efeitos dos fármacos , Erros Inatos do Metabolismo de Esteroides/tratamento farmacológico , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/sangue , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/genética , Criança , Pré-Escolar , Transtorno 46,XY do Desenvolvimento Sexual/sangue , Transtorno 46,XY do Desenvolvimento Sexual/genética , Transtorno 46,XY do Desenvolvimento Sexual/patologia , Esquema de Medicação , Doenças dos Genitais Masculinos/sangue , Doenças dos Genitais Masculinos/genética , Humanos , Hipospadia/sangue , Hipospadia/genética , Hipospadia/patologia , Estudos Longitudinais , Masculino , Proteínas de Membrana/deficiência , Proteínas de Membrana/genética , Mutação , Pênis/crescimento & desenvolvimento , Pênis/patologia , Puberdade/fisiologia , Maturidade Sexual/efeitos dos fármacos , Erros Inatos do Metabolismo de Esteroides/sangue , Erros Inatos do Metabolismo de Esteroides/genética , Erros Inatos do Metabolismo de Esteroides/patologia , Testosterona/sangue , Fatores de Tempo , Resultado do TratamentoRESUMO
Polycystic ovary syndrome (PCOS) is an endocrine disorder with a high prevalence in women of childbearing age. To date, there is no method of efficiently diagnosing PCOS and curing it completely because its pathomechanism remains unclear. Here, we investigated whether metabolic abnormalities maintain the hyperandrogenism and PCOS-like ovaries and whether the symptoms induced by excess androgen are treatable. We ceased the abnormal dihydrotestosterone (DHT) stimulation to determine changes in PCOS-like mice. After ceasing DHT stimulation, the ovarian morphology and gene expression recovered from the DHT-stimulated status. However, after cessation of DHT stimulation, the hypertrophy of adipose tissues and hepatic steatosis were not significantly restored, and fat accumulation-related gene expression and serum metabolic markers in the mice were altered. These findings showed that the reproductive dysfunction was obviously relieved, but because the metabolic abnormalities were not relieved after the cessation of excess androgen for 30 days, it appears that the latter may not maintain the former.
Assuntos
Di-Hidrotestosterona/administração & dosagem , Di-Hidrotestosterona/efeitos adversos , Hiperandrogenismo/induzido quimicamente , Síndrome Metabólica/induzido quimicamente , Síndrome Metabólica/patologia , Síndrome do Ovário Policístico/induzido quimicamente , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/patologia , Animais , Modelos Animais de Doenças , Progressão da Doença , Feminino , Expressão Gênica/efeitos dos fármacos , Hiperandrogenismo/sangue , Hiperandrogenismo/genética , Síndrome Metabólica/sangue , Síndrome Metabólica/genética , Camundongos , Camundongos Endogâmicos C57BL , Ovário/efeitos dos fármacos , Ovário/patologia , Síndrome do Ovário Policístico/sangue , Síndrome do Ovário Policístico/genética , Estimulação Química , Fatores de TempoRESUMO
BACKGROUND: In tissue microarrays, immunostaining of the enzyme arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) was less in recurrent prostate cancers and in cancers with higher Gleason scores. In cultured prostate stem cells, decline in ARSB increased Wnt signaling through effects on Dickkopf Wnt Signaling Pathway Inhibitor (DKK)3. The effects of androgen exposure on ARSB and the impact of decline in ARSB on Wnt signaling in prostate tissue were unknown. METHODS: Epithelial and stromal tissues from malignant and normal human prostate were obtained by laser capture microdissection. mRNA expression of ARSB, galactose-6-sulfate-sulfatase (GALNS) and Wnt-signaling targets was determined by QPCR. Non-malignant human epithelial and stromal prostate cells were grown in tissue culture, including two-cell layer cultures. ARSB was silenced by specific siRNA, and epithelial cells were treated with stromal spent media following treatment with IWP-2, an inhibitor of Wnt secretion, and by exogenous recombinant human Wnt3A. Promoter methylation was detected using specific DKK3 and ARSB promoter primers. The effects of DHT and of ARSB overexpression on DKK expression were determined. Cell proliferation was assessed by BrdU incorporation. RESULTS: Normal stroma showed higher expression of vimentin, ARSB, and Wnt3A than epithelium. Normal epithelium had higher expression of E-cadherin, galactose 6-sulfate-sulfatase (GALNS), and DKK3 than stroma. In malignant epithelium, expression of ARSB and DKK3 declined, and expression of GALNS and Wnt signaling targets increased. In cultured prostate epithelial cells, Wnt-mediated signaling was greatest when ARSB was silenced and cells were exposed to exogenous Wnt3A. Exposure to 5α-dihydrotestosterone (DHT) increased ARSB and DKK3 promoter rmethylation, and effects of DHT on DKK3 expression were reversed when ARSB was overexpressed. CONCLUSIONS: Androgen-induced declines in ARSB and DKK3 may contribute to prostate carcinogenesis by sustained activation of Wnt signaling in prostate epithelium in response to stromal Wnt3A.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/biossíntese , Androgênios/farmacologia , Di-Hidrotestosterona/farmacologia , N-Acetilgalactosamina-4-Sulfatase/biossíntese , Próstata/metabolismo , Neoplasias da Próstata/metabolismo , Proteína Wnt3A/biossíntese , Androgênios/administração & dosagem , Linhagem Celular , Di-Hidrotestosterona/administração & dosagem , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Epitélio/efeitos dos fármacos , Epitélio/metabolismo , Humanos , Microdissecção e Captura a Laser , Masculino , Recidiva Local de Neoplasia/metabolismo , Próstata/efeitos dos fármacos , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Via de Sinalização Wnt/efeitos dos fármacos , Via de Sinalização Wnt/fisiologiaRESUMO
Polycystic ovary syndrome (PCOS) is a continuum of endocrine and reproductive disorders characterized by hyperandrogenism, antral follicle growth arrest, and chronic inflammation. Macrophages play key role in inflammation, and the balance between M1 (inflammatory) and M2 (anti-inflammatory) macrophages determines physiological/pathological outcomes. Here, we investigated if hyperandrogenism increases ovarian chemerin altering the balance of M1 and M2 macrophages and the granulosa cell death. Ovarian chemerin was upregulated by 5α-dihydrotestosterone (DHT) in lean and overweight rats; while increased serum chemerin levels were only evident in overweight rats, suggesting that the serum chemerin may be reflective of a systemic response and associated with obesity, whereas increased ovarian chemerin expression is a localized response independent of the metabolic status. DHT altered follicle dynamics while increased the M1: M2 macrophages ratio in antral and pre-ovulatory follicles. While ovarian M1 macrophages expressing chemokine-like receptor 1 (CMKLR1) were increased, CMKLR1+ monocytes, which migrated toward chemerin-rich environment, were markedly decreased after 15 days of DHT. Androgen-induced granulosa cell apoptosis was dependent on the presence of macrophages. In humans, chemerin levels in follicular fluid, but not in serum, were higher in lean PCOS patients compared to BMI-matched controls and were associated with increased M1: M2 ratio. Our results support the concept that in PCOS, hyperandrogenemia increases chemerin expression while promotes CMKLR1+ monocytes recruitment and deregulates the immunological niche of ovaries. This study established a new immunological perspective in PCOS at the ovarian level. Hyperandrogenism is associated with upregulation of chemerin and macrophage unbalance in the ovaries.
Assuntos
Androgênios/metabolismo , Quimiocinas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Macrófagos/metabolismo , Macrófagos/patologia , Monócitos/metabolismo , Monócitos/patologia , Ovário/metabolismo , Ovário/patologia , Síndrome do Ovário Policístico/metabolismo , Síndrome do Ovário Policístico/patologia , Animais , Apoptose , Movimento Celular/fisiologia , Di-Hidrotestosterona/administração & dosagem , Modelos Animais de Doenças , Feminino , Células da Granulosa/metabolismo , Células da Granulosa/patologia , Humanos , Hiperandrogenismo/metabolismo , Hiperandrogenismo/patologia , Macrófagos/classificação , Ovário/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptores de Quimiocinas/metabolismoRESUMO
Androgen excess is a hallmark of polycystic ovary syndrome (PCOS), a prevalent yet poorly understood endocrine disorder. Evidence from women and preclinical animal models suggests that elevated perinatal androgens can elicit PCOS onset in adulthood, implying androgen actions in both PCOS ontogeny and adult pathophysiology. Prenatally androgenized (PNA) mice exhibit a robust increase of progesterone-sensitive GABAergic inputs to gonadotropin-releasing hormone (GnRH) neurons implicated in the pathogenesis of PCOS. It is unclear when altered GABAergic wiring develops in the brain, and whether these central abnormalities are dependent upon adult androgen excess. Using GnRH-GFP-transgenic mice, we determined that increased GABA input to GnRH neurons occurs prior to androgen excess and the manifestation of reproductive impairments in PNA mice. These data suggest that brain circuit abnormalities precede the postpubertal development of PCOS traits. Despite the apparent developmental programming of circuit abnormalities, long-term blockade of androgen receptor signaling from early adulthood rescued normal GABAergic wiring onto GnRH neurons, improved ovarian morphology, and restored reproductive cycles in PNA mice. Therefore, androgen excess maintains changes in female brain wiring linked to PCOS features and the blockade of androgen receptor signaling reverses both the central and peripheral PNA-induced PCOS phenotype.