Talbott Heather, Delaney Abigail, Zhang Pan, Yu Yangsheng, Cushman Robert A, Cupp Andrea S, Hou Xiaoying, Davis John S
Department of Biochemistry and Molecular BiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5870, USADepartment of Obstetrics and GynecologyOlson Center for Women's Health, University of Nebraska Medical Center, Omaha, Nebraska 68198-3255, USADepartment of Pathology and MicrobiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5900, USAUnited States Department of Agriculture-U.S. Meat Animal Research CenterClay Center, Nebraska 68933-0166, USADepartment of Animal ScienceUniversity of Nebraska-Lincoln, Lincoln, Nebraska 68583-0908, USAVA Nebraska Western Iowa Health Care System and Olson Center for Women's HealthDepartment of Obstetrics and Gynecology, University of Nebraska Medical Center, 983255 Nebraska Medical Center, Omaha, Nebraska 68198-3255, USADepartment of Biochemistry and Molecular BiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5870, USADepartment of Obstetrics and GynecologyOlson Center for Women's Health, University of Nebraska Medical Center, Omaha, Nebraska 68198-3255, USADepartment of Pathology and MicrobiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5900, USAUnited States Department of Agriculture-U.S. Meat Animal Research CenterClay Center, Nebraska 68933-0166, USADepartment of Animal ScienceUniversity of Nebraska-Lincoln, Lincoln, Nebraska 68583-0908, USAVA Nebraska Western Iowa Health Care System and Olson Center for Women's HealthDepartment of Obstetrics and Gynecology, University of Nebraska Medical Center, 983255 Nebraska Medical Center, Omaha, Nebraska 68198-3255, USA.
Department of Biochemistry and Molecular BiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5870, USADepartment of Obstetrics and GynecologyOlson Center for Women's Health, University of Nebraska Medical Center, Omaha, Nebraska 68198-3255, USADepartment of Pathology and MicrobiologyUniversity of Nebraska Medical Center, Omaha, Nebraska 68198-5900, USAUnited States Department of Agriculture-U.S. Meat Animal Research CenterClay Center, Nebraska 68933-0166, USADepartment of Animal ScienceUniversity of Nebraska-Lincoln, Lincoln, Nebraska 68583-0908, USAVA Nebraska Western Iowa Health Care System and Olson Center for Women's HealthDepartment of Obstetrics and Gynecology, University of Nebraska Medical Center, 983255 Nebraska Medical Center, Omaha, Nebraska 68198-3255, USA.
Reproduction. 2014 Jul;148(1):21-31. doi: 10.1530/REP-13-0602. Epub 2014 Mar 31.
Recent studies have suggested that chemokines may mediate the luteolytic action of prostaglandin F2α (PGF). Our objective was to identify chemokines induced by PGF in vivo and to determine the effects of interleukin 8 (IL8) on specific luteal cell types in vitro. Mid-cycle cows were injected with saline or PGF, ovaries were removed after 0.5-4 h, and expression of chemokine was analyzed by qPCR. In vitro expression of IL8 was analyzed after PGF administration and with cell signaling inhibitors to determine the mechanism of PGF-induced chemokine expression. Purified neutrophils were analyzed for migration and activation in response to IL8 and PGF. Purified luteal cell types (steroidogenic, endothelial, and fibroblast cells) were used to identify which cells respond to chemokines. Neutrophils and peripheral blood mononuclear cells (PBMCs) were cocultured with steroidogenic cells to determine their effect on progesterone production. IL8, CXCL2, CCL2, and CCL8 transcripts were rapidly increased following PGF treatment in vivo. The stimulatory action of PGF on IL8 mRNA expression in vitro was prevented by inhibition of p38 and JNK signaling. IL8, but not PGF, TNF, or TGFB1, stimulated neutrophil migration. IL8 had no apparent action in purified luteal steroidogenic, endothelial, or fibroblast cells, but stimulated ERK phosphorylation in neutrophils. In coculture experiments neither IL8 nor activated neutrophils altered basal or LH-stimulated luteal cell progesterone synthesis. In contrast, activated PBMCs inhibited LH-stimulated progesterone synthesis from cultured luteal cells. These data implicate a complex cascade of events during luteolysis, involving chemokine signaling, neutrophil recruitment, and immune cell action within the corpus luteum.
最近的研究表明，趋化因子可能介导前列腺素F2α（PGF）的黄体溶解作用。我们的目的是确定PGF在体内诱导的趋化因子，并在体外确定白细胞介素8（IL8）对特定黄体细胞类型的影响。在发情周期中期的奶牛注射生理盐水或PGF，0.5 - 4小时后切除卵巢，通过qPCR分析趋化因子的表达。在给予PGF后以及使用细胞信号抑制剂分析IL8的体外表达，以确定PGF诱导趋化因子表达的机制。分析纯化的中性粒细胞对IL8和PGF的迁移和激活情况。使用纯化的黄体细胞类型（类固醇生成细胞、内皮细胞和成纤维细胞）来确定哪些细胞对趋化因子有反应。将中性粒细胞和外周血单核细胞（PBMC）与类固醇生成细胞共培养，以确定它们对孕酮产生的影响。在体内PGF处理后，IL8、CXCL2、CCL2和CCL8转录本迅速增加。抑制p38和JNK信号可阻止PGF对体外IL8 mRNA表达的刺激作用。IL8而非PGF、TNF或TGFB1刺激中性粒细胞迁移。IL8在纯化的黄体类固醇生成细胞、内皮细胞或成纤维细胞中无明显作用，但刺激中性粒细胞中的ERK磷酸化。在共培养实验中，IL8和活化的中性粒细胞均未改变基础或LH刺激的黄体细胞孕酮合成。相反，活化的PBMC抑制培养的黄体细胞中LH刺激的孕酮合成。这些数据表明在黄体溶解过程中存在一系列复杂的事件，涉及趋化因子信号传导、中性粒细胞募集以及黄体中的免疫细胞作用。
