Lockwood Charles J, Paidas Michael, Krikun Graciela, Koopman Louise A, Masch Rachael, Kuczynski Edward, Kliman Harvey, Baergen Rebecca N, Schatz Frederick
Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University School of Medicine, 333 Cedar Street, Room 335 FMB P.O. Box 208063, New Haven, Connecticut 06510, USA.
J Clin Endocrinol Metab. 2005 Aug;90(8):4710-5. doi: 10.1210/jc.2004-2528. Epub 2005 May 24.
Decidual neutrophil infiltration complicates spontaneous abortions associated with inflammation and hemorrhage. Transendothelial neutrophil migration into inflamed tissues involves IL-8-mediated chemoattraction, then neutrophil attachment to endothelial cell-expressed intercellular adhesion molecule-1 (ICAM-1).
The aim of this study was to assess IL-8 and ICAM-1 regulation in decidual inflammation and hemorrhage; the effects of the proinflammatory cytokines, TNFalpha, IL-1beta and the hemostatic, proinflammatory cytokine thrombin were measured on IL-8 expression in first trimester decidual cells (DCs), and ICAM-1 immunostaining was compared in normal, inflamed, and hemorrhagic first trimester decidua.
Immunohistochemistry of human decidua and in vitro treatment of human decidual cells were performed.
The study was conducted at the Academic Medical Center.
. DCs were passaged until they were leukocyte-free (CD45 free by FACS), then were incubated with estradiol or medroxyprogesterone acetate alone or with TNFalpha or IL-1beta or thrombin. Normal, inflamed, and hemorrhagic decidua were immunostained for ICAM-1 and the neutrophil marker CD14.
ICAM-1 immunostaining was performed in decidua. IL-8 levels in DC-conditioned media were assessed by ELISA and immunoblotting; IL-8 mRNA levels were measured by quantitative RT-PCR.
Endothelial cell ICAM-1 immunostaining was similar in normal and inflamed or hemorrhagic decidua. In cultured DCs, optimal concentrations of IL-1beta, TNFalpha, and thrombin elevated secreted IL-8 levels by 1083 +/- 261-, 370 +/- 77-, and 45 +/- 15-fold, respectively (mean +/- SEM; P < 0.05; n = 8). The effects were dose dependent and were unaffected by medroxyprogesterone acetate. Western blotting confirmed the ELISA results, and corresponding effects on IL-8 mRNA levels were observed.
Cytokine/thrombin-enhanced DC IL-8 expression interacts with constitutively expressed ICAM-1 in decidual endothelium to modulate neutrophil trafficking into hemorrhagic and inflamed first trimester decidua.
蜕膜中性粒细胞浸润会使与炎症和出血相关的自然流产情况复杂化。中性粒细胞经内皮迁移至炎症组织涉及白细胞介素-8（IL-8）介导的化学趋化作用，随后中性粒细胞黏附于内皮细胞表达的细胞间黏附分子-1（ICAM-1）。
本研究旨在评估蜕膜炎症和出血中IL-8和ICAM-1的调节情况；检测促炎细胞因子肿瘤坏死因子α（TNFα）、白细胞介素-1β（IL-1β）以及具有止血和促炎作用的细胞因子凝血酶对孕早期蜕膜细胞（DCs）中IL-8表达的影响，并比较正常、炎症和出血的孕早期蜕膜中ICAM-1的免疫染色情况。
进行了人蜕膜的免疫组织化学和人蜕膜细胞的体外处理。
该研究在学术医疗中心进行。
DCs传代直至无白细胞（通过流式细胞术检测无CD45），然后单独用雌二醇或醋酸甲羟孕酮或与TNFα、IL-1β或凝血酶一起孵育。对正常、炎症和出血的蜕膜进行ICAM-1和中性粒细胞标志物CD14的免疫染色。
对蜕膜进行ICAM-1免疫染色。通过酶联免疫吸附测定（ELISA）和免疫印迹法评估DC条件培养基中的IL-8水平；通过定量逆转录聚合酶链反应（RT-PCR）测量IL-8 mRNA水平。
正常、炎症或出血的蜕膜中内皮细胞ICAM-1免疫染色相似。在培养的DCs中，IL-1β、TNFα和凝血酶的最佳浓度分别使分泌的IL-8水平升高1083±261倍、370±77倍和45±15倍（平均值±标准误；P<0.05；n = 8）。这些作用呈剂量依赖性，且不受醋酸甲羟孕酮影响。蛋白质免疫印迹法证实了ELISA结果，并观察到对IL-8 mRNA水平的相应影响。
细胞因子/凝血酶增强的DC IL-8表达与蜕膜内皮中组成性表达的ICAM-1相互作用，以调节中性粒细胞向出血和炎症的孕早期蜕膜中的转运。
