Prostaglandin E2 and vascular endothelial growth factor A mediate angiogenesis of human ovarian follicular endothelial cells.
Prostaglandin E2 and vascular endothelial growth factor A mediate angiogenesis of human ovarian follicular endothelial cells.
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DOI：
10.1093/humrep/dev320
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发表时间：
2016-02
影响因子：
6.1
通讯作者：
Heidi A. Trau;M. Brännström;T. Curry;D. Duffy
中科院分区：
文献类型：
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作者：
Heidi A. Trau;M. Brännström;T. Curry;D. Duffy
研究问题 排卵前后，哪些前列腺素 E2 (PGE2) 和血管内皮生长因子 A (VEGFA) 受体介导人类卵泡中的血管生成？摘要答案 PGE2 和 VEGFA 通过多个 PGE2 受体 (PTGER) 和 VEGF 受体 (VEGFR) 发挥作用，在滤泡血管生成中发挥互补作用。已知信息 卵泡产生 PGE2 和 VEGFA 是排卵的先决条件。 PGE2 是一种新兴的血管生成调节剂，尚未在人类排卵卵泡中进行检查。 VEGFA 是滤泡血管生成的既定调节剂。研究设计、大小、持续时间 包含排卵卵泡的卵巢活检取自 11 名接受腹腔镜绝育手术的育龄妇女（30-45 岁）。在某些情况下，女性在卵巢活检前接受 hCG 来替代排卵 LH 激增。此外，从四名接受促性腺激素刺激捐献卵母细胞的育龄妇女（18-31 岁）处获得抽吸物，用于分离人卵巢微血管内皮细胞 (hOMEC)。参与者/材料、环境、方法 利用卵巢活检进行冯维勒布兰德因子的免疫细胞化学检测，以鉴定内皮细胞。 hOMEC 与 PGE2、PTGER 受体选择性激动剂、VEGFA 或 VEGFR 选择性激动剂一起培养。通过 Ki67 免疫细胞化学评估 hOMEC 的增殖情况。 hOMEC 迁移通过对体外多孔膜迁移的细胞进行计数来确定。通过在 3 维基质中培养 hOMEC 后拍摄的照片确定芽的数量和长度来量化芽的形成。主要结果和机会的作用 在 hCG 排卵剂量之前，在人排卵卵泡的颗粒细胞层中未观察到内皮细胞，并且在 hCG 后 18-34 小时首次在颗粒细胞中观察到内皮细胞。在体外，PGE2 增强迁移和芽形成，但不改变 hOMEC 增殖。对每种 PTGER 具有选择性的激动剂增加了迁移，但增殖没有变化。 PTGER1 和 PTGER2 激动剂增加了芽的数量，而只有 PTGER1 影响芽的长度。 VEGFA 增加 hOMEC 的增殖、迁移和类似毛细血管芽的结构的形成。通过 VEGFR1 的信号传导促进 hOMEC 迁移、增殖和少数长内皮细胞芽的形成，而 VEGFR2 刺激则促进 hOMEC 迁移和许多短芽的形成。体外治疗的所有效果均被认为是显着的（P < 0.05）。局限性和注意事项虽然 hOMEC 的原代培养物对 PGE2 和 VEGFA 的反应与其他培养的内皮细胞不同，但 hOMEC 在体内可能不像体外那样对 PGE2 和 VEGFA 做出反应。研究结果的更广泛意义 对 PTGER1、PTGER2、VEGFR1 或 VEGFR2 具有选择性的激动剂和拮抗剂可能具有促进或预防女性排卵的治疗价值。研究经费/竞争利益 这项研究得到了尤尼斯·肯尼迪·施赖弗国家儿童健康和人类发展研究所（HD071875 至 D.M.D.、T.E.C.、M.B.）的拨款资助。作者没有需要披露的利益冲突。
STUDY QUESTION Which receptors for prostaglandin E2 (PGE2) and vascular endothelial growth factor A (VEGFA) mediate angiogenesis in the human follicle around the time of ovulation? SUMMARY ANSWER PGE2 and VEGFA act via multiple PGE2 receptors (PTGERs) and VEGF receptors (VEGFRs) to play complementary roles in follicular angiogenesis. WHAT IS KNOWN ALREADY Production of PGE2 and VEGFA by the follicle are prerequisites for ovulation. PGE2 is an emerging regulator of angiogenesis and has not been examined in the context of the human ovulatory follicle. VEGFA is an established regulator of follicular angiogenesis. STUDY DESIGN, SIZE, DURATION Ovarian biopsies containing the ovulatory follicle were obtained from 11 women of reproductive age (30-45 years) undergoing surgery for laparoscopic sterilization. In some cases, women received hCG to substitute for the ovulatory LH surge before ovarian biopsy. In addition, aspirates from four women of reproductive age (18-31 years) undergoing gonadotrophin stimulation for oocyte donation were obtained for isolation of human ovarian microvascular endothelial cells (hOMECs). PARTICIPANTS/MATERIALS, SETTING, METHODS Ovarian biopsies were utilized for immunocytochemical detection of von Willebrand factor to identify endothelial cells. hOMECs were cultured with PGE2, PTGER receptor selective agonists, VEGFA, or VEGFR selective agonists. hOMECs were assessed for proliferation by Ki67 immunocytochemistry. hOMEC migration was determined by counting cells which migrated through a porous membrane in vitro. Sprout formation was quantified by determining sprout number and length from photographs take after culture of hOMECs in a 3-dimensional matrix. MAIN RESULTS AND THE ROLE OF CHANCE Endothelial cells were not observed within the granulosa cell layer of human ovulatory follicles prior to an ovulatory dose of hCG and were first seen amongst granulosa cells 18-34 h after hCG. In vitro, PGE2 enhanced migration and sprout formation but did not alter hOMEC proliferation. Agonists selective for each PTGER increased migration with no change in proliferation. PTGER1 and PTGER2 agonists increased the number of sprouts, while only PTGER1 affected sprout length. VEGFA increased hOMEC proliferation, migration, and formation of structures resembling capillary sprouts. Signaling through VEGFR1 promoted hOMEC migration, proliferation, and the formation of few, long endothelial cell sprouts, while VEGFR2 stimulation promoted hOMEC migration and the formation of many, short sprouts. All effects of treatments in vitro were considered significant at P < 0.05. LIMITATIONS, REASONS FOR CAUTION While primary cultures of hOMECs respond to PGE2 and VEGFA differently than other cultured endothelial cells, hOMECs may not respond to PGE2 and VEGFA in vivo as they do in vitro. WIDER IMPLICATIONS OF THE FINDINGS Agonists and antagonists selective for PTGER1, PTGER2, VEGFR1, or VEGFR2 may have therapeutic value to promote or prevent ovulation in women. STUDY FUNDING/COMPETING INTERESTS This research was supported by grant funding from the Eunice Kennedy Shriver National Institutes of Child Health and Human Development (HD071875 to D.M.D., T.E.C., M.B.). The authors have no conflicts of interest to disclose.