Smad3基因促进大鼠卵巢颗粒细胞增殖  

Smad3 gene promoting proliferation of GCs in rats

作　　者：金玉洁[1] 董静霞[1] 刘玉霞[1] 申春艳[1] 徐健[1] 

机构地区：[1]北京大学基础医学院人体解剖学与组织胚胎学系,北京100191

出　　处：《解剖学报》2011年第6期766-769,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金面上资助项目(30772344)

摘　　要：目的探讨Smad3基因对大鼠卵巢颗粒细胞增殖功能的影响。方法 21d SD雌性大鼠,腹腔注射孕马血清20IU/只,48h后收集卵巢颗粒细胞进行原代培养,用Smad3基因特异的siRNA基因沉默后,采用免疫细胞化学法检测颗粒细胞中Smad3蛋白表达的变化,以判定基因沉默效率,Smad3基因沉默后用免疫细胞化学法检测增殖细胞核抗原(PCNA)及Cyclin D2蛋白表达的变化。结果 Smad3基因沉默后,卵巢颗粒细胞中PCNA阳性细胞百分比为(25.80±1.70)%,显著低于空白对照组(40.80±2.50)%,Smad3基因沉默后Cyclin D2阳性细胞百分比为(25.00±2.90)%,与空白对照组(55.50±2.50)%相比明显减少。结论 Smad3基因可促进大鼠卵巢颗粒细胞的增殖。Objective To study the effects of Smad3 on the proliferation of the ovarian granulosa cells （GCS） in the rat. Methods Ovarian granulosa cells were cultured and treated in the flollowing conditions： 1. GCs were cultured without any treatment; 2. GCs were cultured with transfection reagent lipofectamine RNAiMAX; 3. GCs were transfected with transfection reagent lipofectamine RNAiMAX and Non-silencing SiRNA; 4. GCs were transfeeted with transfection reagent lipofeetamine RNAiMAX and the SiRNA of Smad3. The expression and localization of PCNA and Cyclin D2 proteins in the four groups were detected by immuoeytochemistry. Results The proteins of PCNA and Cyclin D2 were mainly located in the nuclei of GCs. The percentage of positive cells for PCNA was （25.80 ±1.70） % in the siRNA of Smad3 treated group, which decreased significantly compared to the control group （ P 〈 0.05 ） , and the percentage of the stained cells for Cyclin D2 was （25.00 ± 2.90） % in the siRNA of Smad3 treated group, which decreased significantly compared to the control group（ P 〈 0.05）. Conclusion Smad3 gene could promote the proliferation of ovarian granulosa cells in rats.

关 键 词：SMAD3 SIRNA 颗粒细胞 增殖 免疫组织化学 大鼠 

分 类 号：R322[医药卫生—人体解剖和组织胚胎学]