RESUMO
<p><b>OBJECTIVE</b>To evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill (, WYP) on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism.</p><p><b>METHODS</b>Five groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group (5 in each group) were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f (GTW) either alone (20 mg/kg) or followed by WYP (0.5, 1.0, or 2.0 g/kg daily), respectively for 30 days. Serum levels of follicle-stimulating hormone (FSH), inhibin B (INHB) and testosterone (T) were evaluated using enzyme-linked immunosorbent assay. Androgen-binding protein (ABP) gene expression and transferrin (TF) protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen's score.</p><p><b>RESULTS</b>The toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen's scores were detected in the model group compared with the control group P<0.05 or P<0.01). Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW-induced changes in specific secretory functions of Sertoli cells (ABP, INHB and TF). Furthermore, serum T level and Johnsen's score of the testis increased greatly compared with the model group (P<0.01).</p><p><b>CONCLUSION</b>WYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.</p>
Assuntos
Animais , Masculino , Ratos , Proteína de Ligação a Androgênios , Genética , Metabolismo , Western Blotting , Medicamentos de Ervas Chinesas , Farmacologia , Hormônio Foliculoestimulante , Sangue , Regulação da Expressão Gênica , Inibinas , Sangue , RNA Mensageiro , Genética , Metabolismo , Ratos Sprague-Dawley , Células de Sertoli , Secreções Corporais , Espermatogênese , Comprimidos , Testículo , Biologia Celular , Metabolismo , Testosterona , Sangue , Transferrina , MetabolismoRESUMO
OBJECTIVE@#To determine the effect of soy isoflavones on cell proliferation and the transcription levels of follicle-stimulating hormone receptor (FSHR), inhibin α (INHα), INHβB, androgen binding protein (ABP), transferrin (Tf) and vimentin in testis sertoli cells in SD rats.@*METHODS@#Sertoli cells were cultured in vitro, exposed to daidzein at 0.03, 0.3, 3, and 30 μmol/L and genistein at 0.05, 0.5, 5 and 50 μmol/L, respectively. MTT was used to detect the proliferation of sertoli cells. Real-time PCR was used to detect the relative mRNA expressions of FSHR, INHα, INHβB, ABP, Tf and vimentin.@*RESULTS@#Compared with control groups, cell proliferation and the relative mRNA expression levels of INHβB and ABP in the treated cells showed no significant alternation. The INHα mRNA expression levels were increased in 0.3 and 3 μmol/L Dai and 0.05 μmol/L Gen, while the mRNA expression levels of FSHR were downregulated in 30 μmol/L Dai and Gen at all concentrations. Tf mRNA expression levels were downregulated in 30 μmol/L Dai and 5 μmol/L and 50 μmol/L Gen, and the mRNA expression levels of vimentin were downregulated in 3 and 30 μmol/L Dai and 50 μmol/L Gen.@*CONCLUSION@#Soy Isoflavones may have potential detrimental effect on the male reproductive system, as they may impact the function of sertoli cells by downregulating the transcription levels of some important proteins.
Assuntos
Animais , Masculino , Ratos , Proteína de Ligação a Androgênios , Metabolismo , Subunidades beta de Inibinas , Metabolismo , Inibinas , Metabolismo , Isoflavonas , RNA Mensageiro , Ratos Sprague-Dawley , Receptores do FSH , Metabolismo , Células de Sertoli , Glycine max , Química , Testículo , Biologia Celular , Transferrina , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of different doses of glyphosate on apoptosis and expressions of androgen-binding protein (ABP) and vimentin mRNA in mouse Sertoli cells.</p><p><b>METHODS</b>Primarily cultured mouse Sertoli cells incubated with different doses of glyphosate (60, 90, 120, 150 and 180 mg/L) for 24 h. The growth and morphological alterations in the cells were observed under inverted microscope, and the cell proliferation rate was evaluated withMTT assay. Hoechst 33342 staining was used to detect cell apoptosis after the treatment, and RT-PCR was performed to examine the changes in the expression of ABP and vimentin mRNAs.</p><p><b>RESULTS</b>Sertoli cells exposed to glyphosate showed a reduced cell volume, cell dissociation with occasional cell disruption. The proliferation of the exposed was suppressed with an increased rate of cell apoptosis and lowered expressions of ABP and vimentin mRNAs (P<0.05).</p><p><b>CONCLUSION</b>GLY can cause cellular damages, inhibit cell proliferation, induce cell apoptosis, and decrease expression of ABP and vimentin mRNAs in mouse Sertoli cells in vitro.</p>
Assuntos
Animais , Masculino , Camundongos , Proteína de Ligação a Androgênios , Genética , Metabolismo , Apoptose , Proliferação de Células , Células Cultivadas , Relação Dose-Resposta a Droga , Glicina , Toxicidade , Herbicidas , Toxicidade , RNA Mensageiro , Metabolismo , Células de Sertoli , Biologia Celular , Metabolismo , Vimentina , Genética , MetabolismoRESUMO
<p><b>OBJECTIVE</b>To explore effects of p, p'DDE on the expression of androgen binding protein (ABP), transferrin (Tf) and inhibin B (INH B) mRNA in testis Sertoli cells of Sprague Dawley rats.</p><p><b>METHODS</b>A method has been set up to obtain a large number of viable Sertoli cells from SD rats of 18-20 days of age. With a series of concentration p,p'-DDE (10, 30, and 50 micromol/L) co-incubating the Sertoli cells in vitro, the expression of ABP, Tf and INH B mRNA were determined by RT-PCR.</p><p><b>RESULTS</b>a) With increase of the incubated p, p'-DDE, the expression of ABP mRNA in Sertoli cells went up while that of Tf and INH B dropped in a dose-dependent manner (P < 0. 05). b) The correlation analysis among ABP, Tf and INH B showed that negative relationships were found between ABP and Tf or INH B, respectively (r = - 0. 391 3, P = 0. 032 5; r = - 0.235 2, P = 0.0158), and that positive correlation was indicated between Tf and INH B (r =0.4516, P =0.0047).</p><p><b>CONCLUSION</b>p,p'-DDE is a reproductive toxicant which disrupts the transcription of ABP, Tf and INH B in rat Sertoli cells so as to result in reproductive dysfunction.</p>
Assuntos
Animais , Masculino , Ratos , Proteína de Ligação a Androgênios , Genética , Diclorodifenil Dicloroetileno , Toxicidade , Inibinas , Genética , RNA Mensageiro , Genética , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células de Sertoli , Metabolismo , Transferrina , GenéticaRESUMO
BACKGROUND: We wanted to identify the presence of the estrogen receptor (ER) alpha in Sertoli cells and gain insight on the regulation of the ER alpha gene expression by testosterone in Sertoli cells. The transcriptional regulation of the ER alpha gene was investigated in primary Sertoli cell cultures by in situ hybridization and reverse transcription-polymerase chain reaction (RT-PCR). METHODS: Primary Sertoli cell culture was performed. The expression levels of ER alpha and ER beta mRNA in Sertoli cells were detected by Northern blot, RT-PCR, immunocytochemistry and in situ hybridization. RESULTS: The ovary, testis and epididymis showed a moderate to high expression of ER alpha while the prostate, ovary and LNCap cells showed the ER beta expression. ER alpha mRNA and protein were detected in the germ cells and Sertoli cells by in situ hybridization and immunocytochemistry. The level of ER alpha mRNA was gradually decreased in a time-dependent manner after testosterone treatment, and the changes of ER alpha mRNA were dependent on the concentration of testosterone. Androgen binding protein and testosterone-repressive prostate message-2 (TRPM-2) mRNA were reduced at 24 hour by estradiol, while the transferrin mRNA was not affected. ER alpha mRNA was strongly detectable in the testes of 7 days-old-rats, but it was gradually decreased from 14 to 21 days of age. The primary Sertoli cells also showed the same pattern. The ER alpha gene expression was also regulated by testosterone in the Sertoli cells prepared from the 14- and 21-day old rats. CONCLUSIONS: These results suggest that ER alpha is transcriptionally regulated by testosterone and it may play some role in the Sertoli cells.
Assuntos
Animais , Feminino , Masculino , Ratos , Proteína de Ligação a Androgênios , Northern Blotting , Técnicas de Cultura de Células , Epididimo , Estradiol , Receptor alfa de Estrogênio , Estrogênios , Expressão Gênica , Células Germinativas , Imuno-Histoquímica , Hibridização In Situ , Ovário , Próstata , RNA Mensageiro , Células de Sertoli , Testículo , Testosterona , TransferrinaRESUMO
<p><b>OBJECTIVE</b>The present study was performed to examine functional and structural impairment of rat sertoli cells following dibutyl phthalate (DBP) exposure.</p><p><b>METHODS</b>The 6-week-old healthy male Sprague Dawley rats were randomly divided into 4 groups with 16 animals in each group. DBP dissolved in peanut oil was administered by gavage at doses of 0, 250, 500 and 1 000 mg/kg. After 2-week DBP treatment, half of the rats were sacrificed. The rest were killed following 4-week DBP exposure. Follicle stimulating hormone (FSH) was analysed by radioimmunoassay. The relative expression levels of androgen binding protein (ABP) mRNA and inhibin (INH)alpha mRNA were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The sertoli cell ultrastructures were observed by using transmission electron microscope (TEM).</p><p><b>RESULTS</b>FSH levels were increased after 4-week DBP exposure with significance at doses of 250 and 1 000 mg/kg. Sperm head count and daily sperm product were decreased significantly in 500 and 1 000 mg/kg groups. The expression levels of ABP mRNA were 0.89 +/- 0.15, 0.85 +/- 0.23, 0.54 +/- 0.17, 0.52 +/- 0.16 and 0.88 +/- 0.16, 0.61 +/- 0.12, 0.48 +/- 0.15, 0.47 +/- 0.11 for 0, 250, 500 and 1 000 mg/kg after 2- and 4-week DBP treatments respectively with significance at doses of 500 and 1 000 mg/kg (P < 0.01), while the levels of INHalpha mRNA were 0.88 +/- 0.16, 0.61 +/- 0.12, 0.48 +/- 0.15, 0.47 +/- 0.11 and 0.75 +/- 0.19, 0.56 +/- 0.16, 0.53 +/- 0.08, 0.45 +/- 0.10 with significance at all exposure groups (P < 0.01 or P < 0.05). In sertoli cells of rats exposed to 1 000 mg/kg DBP, TEM photos showed more lysosomes in cytoplasm, proliferated and expanded endoplasmic reticulum and nuclei malformation.</p><p><b>CONCLUSIONS</b>Sertoli cell should be one of the major toxic targets. Impairment of spermatogenesis caused by DBP should be partly due to the suppression of ABP and INHalpha biosynthesis.</p>
Assuntos
Animais , Masculino , Ratos , Proteína de Ligação a Androgênios , Genética , Dibutilftalato , Toxicidade , Relação Dose-Resposta a Droga , Hormônio Foliculoestimulante , Metabolismo , Inibinas , Genética , RNA Mensageiro , Genética , Metabolismo , Radioimunoensaio , Distribuição Aleatória , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células de Sertoli , Metabolismo , Patologia , Testículo , Metabolismo , PatologiaRESUMO
Diabetes is a metabolic disease caused by complicated factors, and its damage to the male reproductive system is threatening men's health. This article reviews the pathophysiological changes in the diabetes-damaged male reproductive system and the mechanism of these changes. Oxidative stress induced by hyperglycemia plays an important role in working damage to the reproductive system of diabetic males, for which some anti-oxidative substances may prove to be an effective cure.
Assuntos
Animais , Humanos , Masculino , Ratos , Proteína de Ligação a Androgênios , Diabetes Mellitus , Patologia , Radicais Livres , Células Intersticiais do Testículo , Metabolismo , Estresse Oxidativo , Células de Sertoli , Secreções Corporais , Testículo , Patologia , TestosteronaRESUMO
The human androgen receptor is a member of the superfamily of steroid hormone receptors and contains three functional domains: an amino-terminal region involved in the expression of androgen regulated genes, a central cystein-rich DNA binding region and a carboxy-terminal hormone binding region. Proper functioning of this protein is a prerequisite for normal male sexual differentiation and development. Androgen action is currently studied in vitro, using fibroblasts culture from genital skin and complementary DNA of the androgen receptor gene has been recently cloned and sequenced. During recent years a substantial progress has been made elucidating the structure-function relationship of the androgen receptor and the characterization of the molecular defects associated with androgen insensitivity syndromes. There appears to be a broad correlation between the degree of receptor dysfunction caused by the mutation and the patient phenotype
Assuntos
Humanos , Masculino , Proteína de Ligação a Androgênios/fisiologia , Receptores Androgênicos/genética , Transtornos do Desenvolvimento Sexual/genética , Di-Hidrotestosterona/farmacocinética , 3-Oxo-5-alfa-Esteroide 4-Desidrogenase/deficiênciaAssuntos
Gravidez , Adulto , Pessoa de Meia-Idade , Humanos , Masculino , Feminino , Androgênios/sangue , Androstenos/metabolismo , Hirsutismo/metabolismo , Proteína de Ligação a Androgênios , Androgênios/metabolismo , Androstano-3,17-diol/sangue , Androstano-3,17-diol/metabolismo , Androstenos/sangue , Desidroepiandrosterona/sangue , Desidroepiandrosterona/metabolismo , Di-Hidrotestosterona/sangue , Di-Hidrotestosterona/metabolismoAssuntos
Gravidez , Adolescente , Adulto , Idoso , Animais , Humanos , Proteína de Ligação a Androgênios/fisiologia , Globulina de Ligação a Hormônio Sexual/fisiologia , Albuminas , Neoplasias da Mama/fisiopatologia , Cirrose Hepática/fisiopatologia , Doença da Mama Fibrocística , Hirsutismo/fisiopatologia , Menopausa/fisiologia , Obesidade/fisiopatologia , Gravidez/fisiologia , Globulina de Ligação a Hormônio Sexual/biossíntese , Esteroides/metabolismo , Testosterona/fisiologia , Testosterona/fisiopatologia , TranscortinaRESUMO
La proteína ligadora de andrógenos (ABP) fue determinada en las fracciones citosólica (cABP) y particulada (pABP), obtenidas por centrifugación diferencial de homogenatos testiculares de ratas. Se prepararon homogenatos en condiciones de estabilización para el ABP por el agregado de testosterona 350 nM al "buffer" de homogeneización. Se observó que tanto los niveles por mg/prot de cABP como de pABP eran máximos entre los 22 y 32 días de edad de la rata, disminuyendo a partir de allí durante la maduración sexual. Cuando los resultados se expresaron por órgano, ambas proteínas aumentaron con la edad del animal. La pABP pudo ser solubilizada en condiciones en que mantuvo su capacidad de unión a andrógenos, procediéndose entonces a su fotomarcación y posterior cromatografía en una columna de Sephadex G-0200, usando el ABP de citosol de epidídimo como control. Se observó que el ABP no sólo comparte con el cABP las mismas características físico-químicas para la unión de andrógenos, sino también el volumen de exclusión en la cromatografía mencionada. Dado que la pABP está solamente presente en la célula de Sertoli, probablemente represente ABP antes de ser secretado. Su localización subcelular sugiere un posible papel del ABP en la distribución de andrógenos testiculares
Assuntos
Ratos , Animais , Masculino , Proteína de Ligação a Androgênios/metabolismo , Citosol/metabolismo , Maturidade Sexual , Testículo/metabolismo , Envelhecimento , Epididimo/metabolismo , Ratos Endogâmicos , Testosterona/metabolismoRESUMO
Se estudió en ratas inmaduras el efecto agudo de la hOG y la testosterona sobre los niveles testiculares (fracciones particulada y soluble) y epididimarios (fracción soluble) de la proteína ligadora de andrógenos (ABP). La administración de una única inyección de hOG (10 UI/rata) produjo una leve disminución de la proteína ligadora de andrógenos (ABP) en el testículo (fracciones particulada y soluble). Este efecto fue observado una y cuatro horas después del tratamiento. Simultáneamente se observó un aumento significativo en la concentración de ABP en el epidídimo (1 h: 169 ñ 5; 4 h: 182 + 5vs. control: 113 ñ13 fmol/mg proteína, media ñ ESM). El aumento en la concentración de testosterona en testículo y epidídimo sugeriría que el efecto observado podría estar mediado por la estimulación gonadatrófica de la síntese de andrógenos en la célula de Leydig. Para comprobar esta hipótesis, el efecto de la hOC se estudió en ratas tratadas previamente con aminoglutetimida para bloquear la esteroidogénesis. En este modelo experimental no se observó un aumento en la concentración de ABP epididimario por la acción de la gonadotrofina. finalmente, la administración de propionato de testosterona indujo un incremento en la concentración de ABP epididimario 4 h después del tratamiento (212 + 18vs. 127 ñ 28 fmol/mg proteína, media ñ ESM). Los resultados obtenidos sugieren que la elevación de los androgénos testiculares inducida por hOG produce un pasaje rápido de ABP desde el testículo hacia el epidídimo
Assuntos
Ratos , Animais , Masculino , Proteína de Ligação a Androgênios/metabolismo , Gonadotropina Coriônica/farmacologia , Epididimo/metabolismo , Testículo/metabolismo , Testosterona/farmacologia , Testosterona/metabolismoRESUMO
Se describe un método de marcación por fotoafinidad de la SHBG sérica humana y de conejo en un precipitado de sulfato de amonio utilizando delta6-testosterona 3H como ligando. La precipitación disminuye la contaminación con albúmina y concentra al mismo tiempo la globulina ligadora de hormonas sexuales. La marcación por fotoafinidad se realizó mediante un flash electrónico. El esteroide libre y el no unido covalentemente se adsorbieron mediante un tratamiento prolongado con una solución de carbón dextrán. La cromatografía en columna de Sephadex G-200 mostró un único pico radioactivo unido covalentemente con el volumen de elución de la SHBG. Este preparado es útil para ser utilizado en estudios de metabolización de la SHBG in vivo. Con algunas modificaciones, el método fue aplicado a la marcación por fotoafinidad del receptor citosólico de andrógenos de próstata utilizando al R 1881 como ligando. El receptor de andrógenos de citosol de próstata también fue precipitado con sulfato de amonio. Luego de marca, irradiar y calentar a 50- C el R 1881-3H no unido covalentemente fue removido mediante el tratamiento con carbón dextrán. En presencia de fotólisis previa al tratamiento con calor, la cromatografía en microcolumnas de Sephadex G-25 mostró un pico radiactivo que eluyó con el volumen de elución de las macromoléculas, el cual desaparecía en las muestras no fotolizadas previamente; sin embargo el receptor fotomarcado tuvo un coeficiente de sedimentación, luego de ultracentrifugación en gradientes lineares de sucrosa, diferente del receptor no irradiado, sugiriendo que la fotólisis generó a algún cambio en la configuración del complejo