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1.
Theriogenology ; 101: 144-150, 2017 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-28708511

RESUMO

Follicle-stimulating hormone (FSH) promotes secretion of follicle fluid and follicle development. FSH acts via cognate FSH receptor (FSHR). It remains unknown whether the supplement of FSH-receptor binding inhibitor (FRBI) into the in vitro maturation (IVM)medium influence the estrogen receptor expression and signal pathway of oocytes in sheep. The present study aimed to investigate FRBI effects on inositol trisphosphate (IP3) of oocytes and protein kinase A (PKA) of sheep granulosa cells, further to elucidate the signal pathway of FRBI effects. Cumulus-oocyte complexes (COCs) were recovered from antral follicles. COCs were cultured for 24 h in the IVM medium supplemented with varying concentrations of FRBI (0, 10, 20, 30 and 40 µg/mL) and FSH (10IU/mL). ELISA was used to measure the concentrations of estradiol (E2) and IP3 in the IVM medium. Western blotting was utilized to detect protein expression of ERß of COCs and protein kinase A (PKA) of granulosa cells. The results showed IP3 concentrations of FRBI-3 and FRBI-4 groups were less than that of CG and FSH groups at 22 h and 24 h (P < 0.05). PKA levels of FRBI-3 and FRBI-4 groups were significantly less than that of CG and FSH group (P < 0.05 or P < 0.01). Expression levels of ERß mRNA and protein of FRBI-treated groups were gradually decreased in comparison to CG and FSH group. The minimum value was detected in the FRBI-4 group. ERß protein level of the FRBI-4 group was significantly less than that of FSH group (P < 0.05). E2 concentrations of FRBI-treated groups were elevated as compared to CG, with the highest increment of FRBI-2 group (P < 0.05). Our results revealed a higher dose of FRBI reduced IP3 production. FRBI could suppress slightly expression levels of ERß mRNA and protein of COCs and PKA of granulosa cells, additionally increased E2 production of sheep COCs.


Assuntos
Proteínas de Transporte/farmacologia , Estradiol/biossíntese , Técnicas de Maturação in Vitro de Oócitos/veterinária , Fragmentos de Peptídeos/farmacologia , Receptores do FSH/genética , Ovinos , Transdução de Sinais/efeitos dos fármacos , Animais , Proteínas de Transporte/administração & dosagem , Meios de Cultura , Meios de Cultivo Condicionados/química , Células do Cúmulo/fisiologia , Proteínas Quinases Dependentes de AMP Cíclico/análise , Estradiol/análise , Receptor beta de Estrogênio/análise , Receptor beta de Estrogênio/genética , Feminino , Hormônio Foliculoestimulante/farmacologia , Expressão Gênica/efeitos dos fármacos , Células da Granulosa/enzimologia , Fosfatos de Inositol/análise , Fosfatos de Inositol/biossíntese , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Fragmentos de Peptídeos/administração & dosagem
2.
Artigo em Chinês | WPRIM | ID: wpr-329052

RESUMO

The design of research protocol and quality control are the key to ensure the quality of clinical trial. A randomized clinical trial regarding the effects of medication combined with acupuncture on live birth rate in polycystic ovary syndrome (PCOS), which was initially designed as a comparative effectiveness research, then added with an acupuncture control group and finally became a factorial analysis, is taken as an example to explain the protocol design and optimization process, demonstrating the high level of methodology design and international recognization. By a series of measurements, such as unified purchase of acupuncture equipment, multiple trainings and assessments for acupuncturists' knowledge and operation standardization, in-site supervision of local center experts, the standard operation of acupuncture could be ensured and the credibility and scientificity of research results could be improved.

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