Department of General & Experimental Pathology, Medical University of Warsaw, ul.Krakowskie Przedmiescie 26/28, Warsaw 00-928, Poland.
Department of Neurology, Second Faculty of Medicine, Medical University of Warsaw, Warsaw, Poland.
J Inflamm (Lond). 2014 May 13;11:12. doi: 10.1186/1476-9255-11-12. eCollection 2014.
Chemokine CX3CL1 possesses unique properties, including combined adhesive and chemotactic functions. Human amniotic epithelial cells (HAEC) show expression of CX3CL1 receptor (CX3CR1) and produce CX3CL1 in response to both physiologic and pathologic stimuli. Chorioamnionitis (ChA) is a common complication of pregnancy and labour. ChA is often accompanied by local hypoxia because of the high oxygen consumption at the site of inflammation. We examined comparatively (ChA-complicated vs. normal pregnancy) CX3CR1 expression and the effects of hypoxia, lipopolysaccharide (LPS), and CX3CR1 blockade on CX3CL1 production in HAEC cultured in vitro.
HAEC have been isolated using trypsinization, and cultured under normoxia (20% O2) vs. hypoxia (5% O2). According to the experimental design, LPS (1 μg/ml) and neutralizing anti-CX3CR1 antibodies were added at respective time points. Mean CX3CL1 concentration in the supernatant samples were determined by ELISA. Expression of immunostained CX3CR1 was analyzed using quantitative morphometry.
We have found that the mean levels of CX3CL1 and CX3CR1 expression were remarkably (p < 0.05) higher in ChA, compared to normal pregnancy. Significantly increased expression of CX3CR1 was observed in ChA during both normoxia and hypoxia. Hypoxia exposure produced decrease in the mean concentration of CX3CL1 in both groups, however this reduction was stronger in normal pregnancy. In normoxia, LPS-evoked rise in the mean concentration of CX3CL1 was higher (p < 0.05) in normal pregnancy. This response was positively correlated with CX3CR1 expression. Blockade of CX3CR1 canceled the secretory response to LPS in all groups.
ChA-complicated pregnancy up-regulates CX3CR1 in HAEC cultured in vitro with simultaneous increase in CX3CL1 production. Hypoxia-resistant production of CX3CL1 may be responsible for ChA-related complications of pregnancy and labor.
趋化因子 CX3CL1 具有独特的特性，包括结合的黏附和趋化功能。人羊膜上皮细胞（HAEC）表达 CX3CL1 受体（CX3CR1），并在生理和病理刺激下产生 CX3CL1。绒毛膜羊膜炎（ChA）是妊娠和分娩的常见并发症。ChA 常伴有局部缺氧，因为炎症部位的耗氧量高。我们比较了（ChA 合并妊娠与正常妊娠）CX3CR1 表达以及缺氧、脂多糖（LPS）和 CX3CR1 阻断对体外培养的 HAEC 中 CX3CL1 产生的影响。
使用胰蛋白酶消化法分离 HAEC，并在常氧（20% O2）与低氧（5% O2）下培养。根据实验设计，在相应时间点加入 LPS（1μg/ml）和中和抗 CX3CR1 抗体。通过 ELISA 测定上清液中 CX3CL1 的平均浓度。使用定量形态计量学分析免疫染色的 CX3CR1 表达。
我们发现，与正常妊娠相比，ChA 中 CX3CL1 和 CX3CR1 表达的平均水平显着升高（p <0.05）。在常氧和低氧条件下，ChA 中观察到 CX3CR1 表达显着增加。低氧暴露导致两组 CX3CL1 的平均浓度降低，但在正常妊娠中降低更明显。在常氧下，LPS 诱导的 CX3CL1 平均浓度升高（p <0.05）在正常妊娠中更高。这种反应与 CX3CR1 表达呈正相关。在所有组中，阻断 CX3CR1 均可消除 LPS 的分泌反应。
体外培养的 HAEC 中 ChA 合并妊娠上调 CX3CR1 并同时增加 CX3CL1 产生。对 CX3CL1 的低氧抵抗性产生可能是导致妊娠和分娩相关 ChA 并发症的原因。
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