Regulation of chemokine production in response to pro-inflammatory cytokines in first trimester decidual cells
Regulation of chemokine production in response to pro-inflammatory cytokines in first trimester decidual cells
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DOI：
10.1016/j.jri.2006.03.002
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发表时间：
2006-12-01
影响因子：
3.4
通讯作者：
Lockwood, C. J.
中科院分区：
文献类型：
--
作者：
Huang, S. J.;Schatz, F.;Lockwood, C. J.
目的：趋化因子通过控制白细胞迁移和淋巴细胞发育来启动免疫反应。蜕膜巨噬细胞的浸润与复发性流产和先兆子痫的发生有关。因此，我们测定了体外培养的人蜕膜细胞是否产生单核/巨噬细胞募集趋化因子，以响应一种强有力的促炎细胞因子--白介素1-β(IL-1β)，以及蜕膜细胞条件培养液中是否含有单核/巨噬细胞趋化活性。方法：无白细胞的早孕蜕膜细胞用雌二醇(E)和醋酸甲羟孕酮(MPA)处理6h，以模拟孕期的类固醇环境，或用E-2和M PA和IL-1β(1 ng/ml)来模拟炎症的蜕膜。以总RNA为模板进行基因合成。制备生物素化的CRNAs，在Affymetrix HG-U133 Plus 2.0芯片上进行化学碎片化杂交，然后进行荧光标记和光学扫描。用GeneSpring7.2软件对Affymetrix GCOS 1.2(基因芯片操作软件)产生的原始数据进行分析。随后，通过实时定量RT-PCR和Western blotting对芯片结果进行验证。结果：IL-1β作用于体外培养的早孕蜕膜细胞后，C-C基序配体2(CCL2)、CCL5、C-X-C基序配体2(CXCL2)、CXCL3和CXCL8等5种单核/巨噬细胞趋化因子的活性从29倍增加到975倍。实时RT-PCR(上调43倍至3069倍)和Western blotting(上调15倍至300倍)的结果证实了核阵列的发现。结论：IL-1β处理早孕蜕膜细胞可诱导单核/巨噬细胞募集趋化因子分泌，促进单核细胞迁移。将这些体外结果外推到植入部位的环境中，表明了IL-1β可能介导蜕膜巨噬细胞过度渗透的机制。(C)2006爱思唯尔爱尔兰有限公司。保留所有权利。
Objective: Chemokines initiate the immune response by controlling leukocyte migration and lymphocyte development. Macrophage infiltration of the decidua has been implicated in the genesis of recurrent miscarriage and preeclampsia. Therefore, we determined whether cultured human decidual cells produce monocyte/macrophage-recruiting chemokines in response to a potent pro-inflammatory cytokine, interleukin-1 beta (IL-1 beta), and whether decidual cell-conditioned medium contains monocyte and macrophage-chemoattractant activity.Methods: Leukocyte-free first trimester decidual cells were treated for 6 h with estradiol (E,) and medroxyprogesterone acetate (MPA) to mimic the steroidal milieu of pregnancy, or E-2 and MPA and IL-1 beta (1 ng/ml) to mimic inflamed decidua. Total RNA was used for cDNA synthesis. Biotinylated cRNAs were generated and chemically fragmented for hybridization on Affymetrix HG-U133 Plus 2.0 chips followed by fluorescence labeling and optical scanning. Raw data generated from Affymetrix GCOS 1.2 (GeneChip Operating Software) were analyzed by GeneSpring 7.2 software. Subsequently microarray results were validated by real time RT-PCR and Western blotting. A functional study of monocyte migration was carried out also using conditioned media from culture.Results: Five chemokines responsible for monocyte/macrophage chemoattraction and activation, including C-C motif ligand 2 (CCL2), CCL5, C-X-C motif ligand 2 (CXCL2), CXCL3 and CXCL8, were markedly elevated from 29- to 975-fold after exposure to IL-1 beta in cultured first trimester decidual cells. The results of real-time RT-PCR (up-regulation from 43- to 3069-fold) and Western blotting (up-regulation from 15- to 300-fold) confirmed the nucroarray findings. Monocyte migration was significantly induced by the conditioned medium from IL-1 beta-treated decidual cells.Conclusions: Treatment of first trimester decidual cells with IL-1 beta induces secretion of monocyte/macrophage recruiting-chemokines and promotes monocyte migration. Extrapolation of these in vitro results to the milieu of implantation site suggests a mechanism whereby IL-1 beta could mediate excessive macrophage infiltration of the decidua. (c) 2006 Elsevier Ireland Ltd. All rights reserved.