Mechanism of Periovulatory Leukocyte Infiltration into the Ovary

排卵期白细胞浸润卵巢的机制

• 批准号: 8609429
• 负责人: CHEMYONG JAY KO
• 金额: $ 13.65万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8609429
• 关键词: Accounting; Adhesions; Adult; Affect; Age; Angiotensins; Animals; Anovulation; Biological Assay; Biological Process; Blood Circulation; Cell Adhesion Molecules; Cell-Adhesion Molecule Receptors; Cells; Chronic; Clinical Treatment; Cytokine Receptors; Cytometry; Data; Defect; Diagnosis; Disease; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Failure; Female; Fertility; Flow Cytometry; Homologous Gene; Human; Immune system; Immunohistochemistry; In Vitro; Indomethacin; Infiltration; Inflammation; Inflammation Mediators; Instruction; Knockout Mice; Lead; Leukocyte Trafficking; Leukocytes; Measures; Mediator of activation protein; Migration Assay; Monitor; Monkeys; Ovarian; Ovarian Stimulations; Ovary; Ovulation; PTGS2 gene; Pathway interactions; Pattern; Plasma; Play; Population; Primates; Procedures; Progesterone; Prostaglandins; RU-486; Rattus; Receptor Cell; Recruitment Activity; Reproductive system; Rodent; Role; Serum; Signal Pathway; Spleen; Testing; Woman; celecoxib; chemokine; cohort; cytokine; inhibitor/antagonist; nonhuman primate; novel; novel strategies; peripheral blood; receptor; reproductive; spatiotemporal

PROJECT SUMMARY (See instructions): Crosstalk between the immune and reproductive systems dramatically impacts fertility. In particular, studies have shown that a tight spatiotemporal control of leukocyte infiltration into the ovary is pivotal for successful ovulation. Once infiltrated, the leukocytes differentiate into a cohort of pro- or anti-infiammatory cells in the preovulatory ovary and play critical roles in ovulation and luteal formation. In this study, we will test the hypothesis that progesterone (P4) and prostaglandins (PG) play pivotal roles in recruiting leukocytes into the preovulatory ovary. Aim 1. To determine the mechanism(s) by which P4 and PG regulate WBC infiltration. We will first determine the roles of P4 and PG on WBC infiltration by treating cycling adult rats with RU486, indomethacin and vehicle, and measuring the effect on the numbers, populations and localization of infiltrating WBC by flow cytometry and immunohistochemistry together with a functional analysis of the expression of chemokines, cytokines, their receptors and cell adhesion molecules (CAMs) in serum, on ovarian cells and WBC by ELISA and qPCR. Aim 2. To determine whether P4 and PG regulate WBC infiltration in primate ovary. In this Aim, we will investigate P4 and PG regulated WBC infiltration in primates. Similar to rodents, primates, including humans also require P4 and PG for normal ovulation. Therefore, we hypothesize that P4 and PG will regulate WBC infiltration in the primate ovary. The actions of P4 and PG will be inhibited by treating primates (Cynomolgous monkeys) with trilostane (3|3HSD inhibitor; inhibits progesterone synthesis), celecoxib (selective COX-2 inhibitor) or vehicle under ovarian stimulation. The effects on the numbers, populations and localization of WBC together with the expression of infiammatory mediators will be determined. Upon the identification of the P4- and PG-regulated WBC subtypes and infiammatory mediators in the monkey, the localization and expression patterns of their homologs will be determined in the human ovary. Aim 3. To determine the regulatory mechanism of preovulatory splenic WBC release. We will identify the trigger(s) and the mechanism of action of splenic WBC release from spleen. We hypothesize that either LH, decreased concentration of circulating WBC or angiotensin-ll (the only known trigger of splenic leukocyte release) act as the trigger. To identify the trigger(s), ovariectomized rats will be treated with candidate triggers (LH and angiotensin-ll) or artificially reduce the concentration of circulating WBC by perfusing rats with plasma. The effects on splenic WBC release will be measured by counting WBC numbers in spleen and peripheral blood.

项目摘要（请参阅说明）： 免疫和生殖系统之间的串扰极大地影响了生育能力。特别是，研究表明，白细胞浸润到卵巢中的紧密时空控制对于成功排卵是关键的。一旦浸润，白细胞将分化成卵巢前的卵巢中的促临时细胞或抗触发细胞，并在排卵和黄体形成中起关键作用。在这项研究中，我们将检验以下假设：孕酮（P4）和前列腺素（PG）在将白细胞招募到卵巢前卵巢中起关键作用。目的1。确定P4和PG调节的机制 WBC浸润。我们将首先通过用RU486，吲哚美辛和媒介物处理循环成年大鼠来确定P4和PG在WBC浸润中的作用，并通过流式细胞仪和不受欢迎的组织的趋势分析，以及他们的comsecins，Cyokines，Cyokines（Cyokines）的表达分析，对WBC的数量，人群和渗透WBC的定位进行衡量，并衡量了趋化的影响。血清，在Elisa和QPCR的卵巢细胞和WBC上。目标2。确定P4和PG是否调节 灵长类动物卵巢中的WBC浸润。在此目的中，我们将研究P4和PG调节灵长类动物的WBC浸润。与啮齿动物类似，包括人在内的灵长类动物也需要P4和PG才能正常排卵。 因此，我们假设P4和PG将调节灵长类动物卵巢中的WBC浸润。 P4和PG的作用将通过用三叶烷（3 | 3HSD抑制剂；抑制孕酮合成），塞来妥（选择性COX-2抑制剂）或卵巢刺激下的赛车抑制剂（抑制孕酮），抑制孕酮合成）来抑制P4和PG的作用。 将确定WBC数量，人群和定位的影响以及感染介体的表达。在鉴定P4和PG调节的WBC亚型以及猴子中的感染介质时，将在人类卵巢中确定其同源物的定位和表达模式。目的3。确定排卵前WBC释放的调节机制。我们将确定脾脏WBC从脾脏释放的触发器和作用机理。我们假设LH，循环WBC浓度降低或血管紧张素-LL（唯一已知的脾白细胞释放的触发器）作为触发。为了识别触发因素，将用候选触发器（LH和血管紧张素-LL）处理卵巢切除的大鼠，或者通过用血浆灌注大鼠，从而人为地降低循环WBC的浓度。对脾脏WBC释放的影响将通过计算脾和外周血中的WBC数来衡量。