摘要
目的:研究卵泡刺激素(FSH)对大鼠卵巢颗粒细胞中Smad2/Smad3蛋白的调节。方法:21dSD雌鼠,腹腔注射孕马血清20IU,对颗粒细胞进行原代培养,用转化生长因子β受体Ⅱ(TβRⅡ)抗体中和卵巢颗粒细胞表面TβRⅡ,以阻断TGFβ/Smads信号通路,再以不同浓度的FSH对细胞进行不同时间的处理,免疫印迹法检测TGFβ信号通路中TβRⅡ、Smad2/Smad3及P-Smad2/P-Smad3蛋白的表达。结果:TβRⅡ、Smad2/Smad3、P-Smad2/P-Smad3蛋白的表达量与FSH的作用呈时间依赖性显著增强;TGFβ/Smads信号通路被阻断后再用FSH处理,Smad2/Smad3、P-Smad2/P-Smad3蛋白的表达量没有明显变化;细胞表面TβRⅡ恢复后,Smad2/Smad3、P-Smad2/P-Smad3蛋白的表达量有少量增加。结论:FSH刺激大鼠卵巢颗粒细胞中Smad2/Smad3的表达和磷酸化;FSH对Smad2/Smad3的激活作用在相当程度上依赖于TβRⅡ。
Objective: To study the effects of follicle stimulating hormone (FSH) on the expression and phosphorylation of Smad2/Smad3 proteins and the interaction between FSH and TGF-β/Smads signaling pathway in rat ovarian granulosa cells (GCs). Methods:The GCs from female SD rats of 21 d were cultured with TβRⅡ antibody and FSH at different concentration, and then cells were obtained at defined points to determine the expression of Smad2/Smad3 and P-Smad2/P-Smad3 by Western Blot assay. Results: FSH enhanced the expression of TβRⅡ, Smad2/Smad3 and P-Smad2/P-Smad3 proteins in manner of time dependent; Neutralization of TβRⅡ antibody to TβRⅡ on the surface of GCs inhibited the stimulation of FSH on Smad2/Smad 3 in GCs; The effects of FSH on Smad2/Smad3 proteins recovered when TβRⅡ was expressed again. Conclusion: In the culture condition, FSH stimulated the expression and phosphorylation of Smad2/Smad3; The activation of FSH on Smad2/Smad3 depended on TβRⅡ.
出处
《解剖学杂志》
CAS
CSCD
北大核心
2008年第2期141-144,共4页
Chinese Journal of Anatomy
基金
北京大学医学部985基金(612206062)
教育部留学回国基金(413152015)