The effect of conditioned medium derived from human placental multipotent mesenchymal stromal cells on neutrophils: possible implications for placental infection
The effect of conditioned medium derived from human placental multipotent mesenchymal stromal cells on neutrophils: possible implications for placental infection
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DOI：
10.1093/molehr/gau062
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发表时间：
2014-11-01
影响因子：
4
通讯作者：
Wu, Yi-Hsin
中科院分区：
文献类型：
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作者：
Chen, Chie-Pein;Chen, Yi-Yung;Wu, Yi-Hsin
人胎盘多能间充质基质细胞（hPMSCs）在胎盘炎症中的作用尚不清楚。我们假设，人PMSCs参与胎盘感染的早期阶段。从足月胎盘中分离hPMSC，从外周血中分离嗜中性粒细胞。采用RT-PCR、流式细胞术和酶联免疫吸附试验检测hPMSCs中Toll样受体（TLR）和细胞因子的表达。通过流式细胞术和western blot检测脂多糖（LPS）预处理和未预处理的hPMSCs条件培养液对中性粒细胞迁移、凋亡和活性氧（ROS）产生的影响。hPMSC表达TLR 1、TLR 3、TLR 4、TLR 6、TLR 7和TLR 9。LPS刺激增加hPMSC TLR 4的表达以及IL-6和IL-8的产生。中性粒细胞对hPMSC条件培养基表现出趋化性，其被IL-8耗尽抑制。hPMSC条件培养液可促进神经元CD 11b活化，而LPS预处理的hPMSC培养液可进一步增强神经元CD 11b活化。hPMSC条件培养基减少嗜中性粒细胞ROS产生。LPS单独增加了中性粒细胞的吞噬作用，但不增加有或没有LPS刺激的hPMSC条件培养基。hPMSC条件培养基诱导中性粒细胞中的STAT 3活化，其被针对IL-6的中和抗体抑制。hPMSC条件培养基拯救中性粒细胞免于凋亡，但这种作用在LPS预处理的hPMSC条件培养基中显著降低。从条件培养基中消耗IL-6进一步抑制对中性粒细胞的抗凋亡作用。我们的研究结果表明，hPMSCs可以与外周血中性粒细胞相互作用，以响应胎盘的炎症信号。hPMSC产生的细胞因子可诱导中性粒细胞趋化并减少中性粒细胞凋亡。
The role of human placental multipotent mesenchymal stromal cells (hPMSCs) in placental inflammation is unknown. We hypothesize that hPMSCs are involved in the early phases of placental infection. hPMSCs were isolated from term placentas and neutrophils from peripheral blood. The expression of toll-like receptors (TLRs) and cytokines by hPMSCs was determined by RT-PCR, flow cytometry and enzyme-linked immunosorbent assay. The effect of conditioned medium of hPMSCs with or without lipopolysaccharide (LPS) pretreatment on neutrophil functions: migration, apoptosis and production of reactive oxygen species (ROS) was assessed by flow cytometry and western blot. hPMSCs expressed TLR1, TLR3, TLR4, TLR6, TLR7 and TLR9. LPS stimulation increased the expression of TLR4 and the production of IL-6 and IL-8 by hPMSCs. Neutrophils exhibited chemotaxis to hPMSC-conditioned medium, which was inhibited by IL-8 depletion. Neutrophil CD11b activation was promoted by hPMSC-conditioned medium, which was further enhanced in media from hPMSCs pretreated with LPS. hPMSC-conditioned medium reduced neutrophil ROS production. Neutrophil phagocytosis was increased by LPS alone but not by hPMSC-conditioned medium with or without LPS stimulation. hPMSC-conditioned medium induced STAT3 activation in neutrophils, which was inhibited by neutralizing antibody to IL-6. hPMSC-conditioned medium rescued neutrophils from apoptosis, but this effect was significantly reduced in conditioned medium of hPMSCs with LPS pretreatment. Depletion of IL-6 from the conditioned medium further inhibited the anti-apoptotic effect on neutrophils. Our results demonstrate that hPMSCs can interact with peripheral blood neutrophils in response to inflammatory signals of the placenta. Cytokines produced by hPMSCs can induce neutrophil chemotaxis and reduce neutrophil apoptosis.