FSH和胰岛素对绵羊卵巢卵泡颗粒细胞体外培养的影响  

Effects of FSH and Insulin on Sheep Ovarian Follicular Granulosa Cells in vitro Culture

作　　者：李鹏飞[1] 岳文斌[2] 庞钰莹[2] 于雪静[2] 黄洋[2] 任有蛇[2] 吕丽华 

机构地区：[1]山西农业大学生命科学学院,太谷030801 [2]山西农业大学动物科技学院,太谷030801

出　　处：《畜牧兽医学报》2013年第9期1386-1391,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31172211);农业部948项目(2010-Z43);山西省留学基金项目(2008080);山西省国际科技合作项目(2010081002);山西农业大学新增横向协作与委托项目(2010HX54);国家重点实验室开放课题(SNKF-2012-02);山西省科技攻关项目(20130311027-2)

摘　　要：旨在探讨在无血清体外培养条件下,促卵泡素(Follicular stimulating hormone,FSH)和胰岛素对绵羊卵巢卵泡颗粒细胞增殖和雌激素分泌的影响。收集卵巢卵泡颗粒细胞,Long-term培养方法在FSH、胰岛素不同浓度下培养168h后,在显微镜下观察各孔细胞生长情况并计数;采用双抗体ABC-ELISA法测定雌激素浓度。结果表明,FSH与胰岛素的共培养体系,颗粒细胞的增殖和雌激素的分泌量显著高于各单独处理组;胰岛素浓度为10ng·mL-1,FSH浓度分别为5.0和25.0ng·mL-1时,2组颗粒细胞数最高,但差异不显著;当胰岛素浓度为10ng·mL-1,FSH浓度为5.0ng·mL-1时,雌激素分泌量最高,与其他组差异显著(P<0.05),此时颗粒细胞的生长状况最好,培养体系最佳。绵羊卵巢卵泡颗粒细胞增殖和雌激素分泌需在FSH诱导下进行;胰岛素对FSH诱导的绵羊卵巢卵泡颗粒细胞增殖和雌激素的分泌有促进作用;二者对颗粒细胞增殖和雌激素的分泌存在剂量效应关系。The study focused on the effect of FSH and insulin on the proliferation of granulosa cells and the production of estradiol when the sheep follicle granulosa cells were cultured in ser- um-free medium. Granulosa cells were collected and treated with a range of concentration of FSH and insulin, and cultured for 168 h using long term method. Observed and counted under the mi- croscope. Estradiol concentration was measured by double antibody ABC-ELISA. Results indica- ted that granulosa cell proliferation and estradiol production in the groups treated with FSH and insulin were superior to that in other groups treated alone. At 10 ng .mL-1 concentration of insulin and 5.0 or 25.0 ng . mL-1 concentration of FSH in medium, the number of granulosa cells were most and there is no significant difference between these two treated groups; at 10 ng . mL-1 concentration of insulin and 5.0 ng .mL-1 concentration of FSH in medium, Estradiol concentration was the highest （P〈0.05） and the culture system was optimal. The multiplication and secretion of estradiol insheep ovarian granulosa cells were induced by FSH. Insulin promotes the multiplication and es- tradiol secretion of FSH-induced sheep follicular granulosa cells in vitro. FSH and insulin stimu- late the multiplication and estradiol secretion of sheep follicular granulose cells in a concentration- dependent manner.

关 键 词：绵羊 颗粒细胞 体外培养 促卵泡素 胰岛素 

分 类 号：S826[农业科学—畜牧学] S814.1[农业科学—畜牧兽医]