胰岛素和FSH对体外培养猪卵泡颗粒细胞雌激素的影响  

Effects of Insulin and FSH on E2Production of Pig Ovarian Follicular Granulosa Cells in vitro Culture

作　　者：李鹏飞[1] 孟金柱 郝庆玲 毕锡麟 王锴[3] 朱芷葳[1] 吕丽华 

机构地区：[1]山西农业大学生命科学学院,太谷030801 [2]铜仁学院乌江学院,铜仁554300 [3]山西农业大学动物科技学院,太谷030801

出　　处：《畜牧兽医学报》2017年第11期2084-2090,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31402156);山西省回国留学人员科研资助项目(2014-重点5);山西省国际科技合作项目(201603D421006);山西省三晋学者和人才引进项目;山西农业大学创新基金项目(zdpy201403/201503);引进人才博士科研启动基金(2014ZZ04);青年拔尖创新人才支持计划(TYIT201403);山西省重点研发计划(一般)农业项目(201703D221020-1)

摘　　要：旨在研究胰岛素(Insulin)和促卵泡素(Follicular stimulating hormone,FSH)对猪卵泡发育过程的影响。猪屠宰后收集卵巢,选择健康卵泡并分离颗粒细胞(Granulesa cells,GCs),设定对照组和重复组,胰岛素、FSH浓度梯度下Long-term体外培养,7d后显微镜观察细胞增殖情况并计数,竞争法测定雌激素(E_2)浓度。数据分析表明:当FSH为0ng·mL^(-1)时,随着胰岛素浓度的增加,猪卵泡GCs细胞密度增加,细胞计数显示GCs数量呈上升趋势,且胰岛素浓度为100ng·mL^(-1)时,细胞数量最高(P<0.05);当胰岛素为0ng·mL^(-1)时,随着FSH浓度的增加,细胞密度增加,细胞计数显示GCs数量呈上升趋势,且FSH浓度为5和25ng·mL^(-1)时,细胞数量显著高于其它两组(P<0.05);E_2测定结果显示,当FSH为0ng·mL^(-1),胰岛素为100ng·mL^(-1)时,培养液E_2浓度显著高于他不同浓度胰岛素组(P<0.05);当FSH为1ng·mL^(-1),胰岛素为10ng·mL^(-1)时,培养液E2浓度最高,但与不同浓度胰岛素各组差异不显著(P>0.05);FSH浓度为5和25ng·mL^(-1)时,不同浓度胰岛素各组之间E_2浓度差异不显著。猪卵泡发育过程中,胰岛素和FSH均有促颗粒细胞增殖和E_2分泌的能力,FSH超过一定生理浓度会降低卵泡颗粒细胞分泌E_2。To research the effects of insulin and FSH on porcine ovarian follicular development, ovaries were collected and follicular granulosa cells （GCs） were isolated. Setting control groups and repetitive groups, GCs were cultured for 7 d treated with different concentrations of insulin and FSH by long-term culture in vitro. Observing the growth situation and counting the numbers of GCs in each well. Competition method was performed to detect the E2 concentrations in medi- um. Results showed that, at 0 ng·mL 1concentration of FSH in medium, with the increase of insulin concentration, GCs proliferation was more and the number of GCs showed a tendency to increase. The numbers of GCs were the most when the supplement dose of insulin was 100ng·mL^-1 （P〈0.05）. When insulin was 0ng·mL^-1, with the increase of FSH concentration, GCs prolif- eration was more and the number of GCs showed a tendency to increase, and at 5 or 25ng·mL^-1 concentration of FSH, the number of GCs were more than other 2 groups （P〈0.05）. When FSH at 0 ng ·mL^-1, insulin at 100 ng·mL^-1, E2 concentration was more than other groups with different concentration of insulin （P〈0.05）. When insulin was 10 ng·mL^-1 and FSH was 1ng·mL^-1, E2 concentration in the medium was the greatest, but there was no significant differences among groups with different concentrations of insulin （P〈0. 05）. At 5 or 25 ng·mL^-1 concentration of FSH, there was no significant differences in E2 concentration among different groups with different concentrations of insulin. Insulin in cooperation with FSH promotes the proliferation and E2 production of porcine follicular GCs. The E2 secretion of porcine follicular GCs will decline when FSH exceeded a certain physiological concentration.

关 键 词：猪 颗粒细胞 胰岛素 促卵泡素 雌激素 体外培养 

分 类 号：S828.2[农业科学—畜牧学]