山羊卵巢间质细胞对小腔卵泡发育的影响  

Effect of Ovarian Stromal Cells on Co-cultured Small Antral Follicles in the Goat

作　　者：邱明宁[1] 韩成全[1] 杨忠财[1] 刘军[1] 权富生[1] 张涌[1] 

机构地区：[1]西北农林科技大学动物医学院,农业部动物生物技术重点开放实验室,杨凌712100

出　　处：《畜牧兽医学报》2013年第7期1043-1050,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金："十二五"农村领域国家科技计划课题;"863"项目(2011AA100303)

摘　　要：为了能有效利用山羊小腔卵泡和研究相关小腔卵泡发育的机理,将山羊小腔卵泡与卵巢皮质间质细胞、卵泡膜细胞和卵巢髓质间质细胞体外共培养,测定培养第5天卵泡直径、培养液中类固醇激素分泌量、卵母细胞特异性分泌因子gdf9和bmp15基因的表达和卵泡颗粒细胞以及卵母细胞凋亡与抗凋亡基因表达。研究结果显示,山羊小腔卵泡与卵巢间质细胞共培养5d,卵泡直径增加明显大于小腔卵泡单独培养组(P<0.05);小腔卵泡与卵巢皮质间质细胞共培养5d后,小腔卵泡分泌的雌二醇激素的量高于小腔卵泡单独培养组(P<0.01),为413.95pg·mL-1;小腔卵泡与卵巢髓质间质细胞共培养5d后,小腔卵泡分泌孕酮的量(11.695ng·mL-1)极显著高于小腔卵泡单独培养组和其他共培养组(P<0.01),小腔卵泡与卵泡膜细胞共培养5d后,小腔卵泡分泌的孕酮的量高于小腔卵泡单独培养组(P<0.05);小腔卵泡与卵巢皮质间质细胞、髓质间质细胞以及与卵泡膜细胞共培养5d后,其卵母细胞特异性分泌因子gdf9基因和bmp15基因表达显著上调(P<0.01);与卵巢间质细胞共培养5d后,小腔卵泡内卵丘细胞和卵母细胞抗凋亡基因表达上调(P<0.05),小腔卵泡内卵丘细胞和卵母细胞抗凋亡基因表达下调(P<0.05)。表明卵巢间质细胞促进小腔卵泡的生长以及类固醇激素如雌二醇和孕酮分泌,促进小腔卵泡内卵母细胞特异性分泌的生长因子(gdf9和bmp15)基因的表达,同时抑制小腔卵泡的凋亡。尤其是卵巢卵泡膜细胞与小腔卵泡共培养后明显提高小腔卵泡的抗凋亡能力。To investigate the mechanism of follicular development in the goat, our present study were designed to co-culture small antral follicles with ovarian cortex stromal cells, theca cells and ovarian medullary stromal cells in vitro for 5 d, respectively, and then to measure the change of diameter of follicles, steroid hormone productions, expressions of oocyte secreted factors （gdf9 and bmp15） and inhibitory apoptotic and pro-apoptotic genes. The results showed that the diameter of follicles were increased significantly in co-cultured groups than which cultured alone after 5 d culture （P〈0.05） ; estradiol production increased in small antral follicles （413.95 pg ·mL-1） after co-cultured with ovarian cortical stromal cells for 5 d （P〈0.01）; progesterone production extremely increased in small antral follicles （11. 695 ng · mL-1） when co-cultured with ovarian medullary stromal cells for 5 d （P〈0.01）, progesterone production increased in small antral follicles when co-cultured with theca cells for 5 d （P〈0. 05）; expressions of gdf9 and bmp15 mRNA in the oocytes in follicles co-cultured with ovarian stromal ceils and theca ceils were increased （P〈0. 01）; expressions of inhibitory apoptotic genes in cumulus cells and oocytes in small antral follicles co-cultured increased （P%0.05）, and expressions of pro-apoptotic genes in cumulus cells and oocytes in small antral follicles co-cultured decreased （P〈0.05）. These results indicated that ovarian stromal cells and theca cells could promote growing of small antral follicles, steroid hormone productions, and expressions of oocyte secreted factors （gdf9 and bmp15）, and inhibit apoptosis of small antral follicles; especially enhance the capacity of small antral follicles significantly through inhibiting apoptosis after co-cultured with theca cells.

关 键 词：小腔卵泡 卵巢间质细胞 共培养 细胞凋亡 山羊 

分 类 号：S827[农业科学—畜牧学] S814.1[农业科学—畜牧兽医]