摘要
目的研究Smad2/Smad3蛋白在大鼠卵巢颗粒细胞中的表达及FSH对其活化的影响。方法21dSD雌性大鼠,注射PMSG 20IU,48h后对卵巢颗粒细胞进行原代培养,用TGFβRⅡ抗体及不同浓度的FSH对细胞进行不同时间的处理,通过免疫细胞化学方法观察TGFβ信号通路中Smad2/Smad3和P-Smad2/P-Smad3(磷酸化Smad2/3)表达的变化。结果1.Smad2/Smad3主要定位于卵巢颗粒细胞胞质,其活化形式P-Smad2/P-Smad3有少量表达;2.经FSH处理后,Smad2/Smad3向核内转移增多,P-Smad2/P-Smad3的表达增强,并与FSH的作用时间及剂量呈正相关性;3.TGFβRⅡ被抗体完全中和后再用FSH处理,Smad2/Smad3的核阳性率无显著增多,P-Smad2/P-Smad3的表达未见明显增强。结论1.FSH促进Smad2/Smad3的磷酸化;2.FSH对Smad2/Smad3的激活作用与TGFβ受体密切相关。
Objective To study the effects of FSH on the phosphorylation of Smad2/Smad3 protein in rat ovarian granulosa cells. Methods The granulosa cells of 21-day-old SD female rats were cultured with TGFβRⅡ antibody and FSH of different concentrations, and then the cells were obtained at defined points to determine the phosphorylation of Smad2/Smad3 by immunocytochemistry. Results 1. Both Smad2 and Smad3 proteins were mainly expressed in the cytoplasm of ovarian granulosa cells,and a small amount of phosphorylated-Smad2/3 were expressed in the nucleus; 2. The treatment with FSH increased the nuclear transfer of Smad2/Smad3 and the expression of P-Smad2/P-Smad3 in granulosa cells, The effects of FSH were time and dose dependent; 3. The antibody-neutlised TGFβRⅡ inhibited the stimulation of FSH on Smad2/3 in granulosa cells. Conclusion 1. FSH stimulates the phosphorylation of Smad2/Smad3; 2.The activation of FSH on Smad2/Smad3 is depends on TGFβRⅡ.
出处
《解剖学报》
CAS
CSCD
北大核心
2007年第2期205-208,共4页
Acta Anatomica Sinica
基金
北京大学医学部985基金(612206062)
教育部留学回国基金(413152015)
