LUTEINIZED HUMAN GRANULOSA CELLS VASCULAR ENDOTHELIAL GROWTH FACTOR PRODUCTION

最近的证据表明，VEGF，一种由人类产生的细胞因子， 颗粒细胞，是卵巢发育的致病因子 过度刺激综合征（OHSS）。 本研究的目的是 确定是否在体外暴露于黄体化颗粒细胞 促性腺激素（hCG）刺激VEGF的产生，如果孕酮（P） 由这些细胞产生的是VEGF表达的自分泌调节剂。 黄体化的颗粒细胞收集自接受 在7500 IU hCG推注后34小时进行卵巢刺激进行IVF。 在研究1中，将细胞在DMEM/F12培养基中培养9天 （对照）补充hCG（CR 123，100 ng/ml），低密度 脂蛋白（LDL，100 fg蛋白/ml）或hCG + LDL。 在研究2中，P 合成抑制剂，曲洛司坦（Sanofi温斯洛普，科尔奇维尔，PA）， 和/或P受体拮抗剂ZK 137.316（Schering AG，柏林， 德国）添加到选定的威尔斯孔中。 测定培养基中的VEGF （ELISA; R&D Systems，Minneapolis，MN）。 研究1的结果是 基于第1天的对照VEGF水平分为两类 “生产者”（n=6; <750 pg VEGF/ml）和“高生产者”（n=5; >1000 pg VEGF/ml VEGF/ml）。 低和高生产者培养物中的VEGF水平 从第1天增加（p<0.01）至第3天的最大水平，当它们 在培养的第9天稳定下降。 VEGF水平低 在第3天用LDL处理后，生产者增加（p<0.05）， 5、7和9，并在培养的第5天和第7天用hCG。 的影响 hCG和LDL联合使用与单独使用LDL的结果相似。 HCG也 到第7天，高生产者的VEGF产量增加（p<0.05）。 在研究2中，曲洛司坦处理降低了（p<0.05）P的产生， 在培养的第1天，91%，但对任何细胞的VEGF产生都没有影响。 天 此外，曲洛司坦/ZK联合治疗对小鼠的生长没有影响。 观察VEGF合成。 这些结果表明，VEGF LGC的产生受促性腺激素（hCG）直接调节 独立于促性腺激素刺激的孕酮合成。 这些 数据与OHSS恶化的假设一致 在妊娠早期是由CG刺激VEGF介导的 生产

Recent evidence suggests that VEGF, a cytokine produced by human granulosa cells, is a causative factor in the development of ovarian hyperstimulation syndrome (OHSS). The objective of this study was to determine if in vitro exposure of luteinized granulosa cells to gonadotropin (hCG) stimulates VEGF production and if progesterone (P) produced by these cells is an autocrine regulator of VEGF expression. Luteinized granulosa cells were collected from women undergoing ovarian stimulation for IVF, 34 hours after a 7500 IU bolus of hCG. In Study 1, cells were cultured for 9 days in DMEM/F12 medium (control) supplemented with hCG (CR123, 100 ng/ml), low density lipoprotein (LDL, 100 fg protein/ml), or hCG + LDL. In Study 2, the P synthesis inhibitor, trilostane (Sanofi Winthrop, Collegeville, PA), and/or a P receptor antagonist, ZK137.316 (Schering AG, Berlin, Germany) were added to selected wells. Medium was assayed for VEGF (ELISA; R&D Systems, Minneapolis, MN). Results of Study 1 were divided in two categories based on control VEGF levels on day 1 "low producers" (n=6; <750 pg VEGF/ml) and "high producers" (n=5; >1000 pg VEGF/ml). VEGF levels in cultures of both low and high producers increased (p<0.01) from day 1 to maximal levels on day 3, when they steadily declined through day 9 of culture. VEGF levels of low producers increased (p<0.05) following treatment with LDL on days 3, 5, 7 and 9 and with hCG on days 5 and 7 of culture. The effects of combined hCG and LDL were similar to those of LDL alone. HCG also increased (p<0.05) VEGF production for the high producers by day 7. In Study 2, trilostane treatment decreased (p<0.05) P production by 91% on day 1 of culture, but had no effect on VEGF production on any day. Moreover, no effect of the combined trilostane/ZK treatment was observed on VEGF synthesis. These results suggest that VEGF production by LGC is directly regulated by gonadotropin (hCG) independent of gonadotropin-stimulated progesterone synthesis. These data are consistent with the hypothesis that the exacerbation of OHSS in early pregnancy is mediated by the CG stimulation of VEGF production.