流式细胞计数测定抗孕激素对猪卵巢细胞孕酮受体的影响  

Effect of antiprogestin in progesterone receptor on porcine ovarian cells by flow cytometry

作　　者：吴尔若 王建中[2] 赵金来 

机构地区：[1]国家计划生育委员会科学技术研究所,北京100081 [2]北京医科大学第一医院

出　　处：《生殖医学杂志》2000年第2期77-82,共6页Journal of Reproductive Medicine

摘　　要：为了研究抗孕激素调节卵巢功能的机理 ,制备了人胎盘羊膜的双池培养系统。分离猪卵巢颗粒细胞和内泡膜细胞 ,分别种在羊膜两侧。在加入或不加 RU486和ZK98.73 4时 ,两种细胞在促性腺激素刺激下共同培养。 48h后 ,应用流式细胞计数(FCM)测定两种细胞的孕酮受体 (PR)和雌激素受体 (ER) ;放射免疫法测定培养液中的孕酮 (P)和雌二醇 (E2 )浓度。结果显示 ,抗孕激素增加两种细胞的 PR含量 ,ER没有明显改变 ;颗粒细胞的 P和 E2 分泌受到抑制 ,内泡膜细胞的 P分泌也减少。提示 P调节 P本身的合成 ,这种旁分泌和自分泌作用是通过 P控制 PR上调的途径。Objectives: To investigate the effects of antiprogestin on the ovarian steroidiogenesis and the regulation of steroid receptors. Methods: Flow cytometry (FCM) was employed to detect progesterone receptor (PR) and estrogen receptor (ER) of porcine granulosa and thecal cells. Both types of cells were incubated in a dual chamber system under the stimulation of gonodotropins with or without RU486 or ZK98.734. This culture system was prepared with the amnion membrane of human placenta. After 48 h incubation PR and ER contents of both cells were detected by FCM. The concentrations of progesterone (P) and estradiol (E 2) in the media were measured by radioimmunoassay. Results: Both antiprogestins increased PR contents of thecal cells in doses of 30 nmol/L and 3 000 nmol/L. For granulosa cells, the PR contents were increased by both antiprogestins in the dose of 3 000 nmol/L. In the dose of 30 nmol/L, only ZK98.734 up regulate PR contents of granulosa cells. No significant change was found for ER in the present study. At the same time the productions of P and E 2 by granulosa cells were inhibited, the P production by thecal cells was reduced also. Conclusion: These data suggest that P regulates P synthesis in ovarian steroidiogenesis. It may be that progesterone controls PR up regulation through the autocrine/paracrine action. It could be the mechanism of action of antiprogestin on the development and steroidogenic function of ovarian follicles.

关 键 词：黄体酮受体 雌二醇受体 卵巢 粒层细胞 卵泡膜细胞 孕酮受体 流式血细胞计数 双池培养 抗孕激素 

分 类 号：R711.75[医药卫生—妇产科学] R339.22[医药卫生—临床医学]