Unfractionated Heparin Modulates Lipopolysaccharide-Induced Cytokine Production by Different Signaling Pathways in THP-1 Cells
Unfractionated Heparin Modulates Lipopolysaccharide-Induced Cytokine Production by Different Signaling Pathways in THP-1 Cells
复制标题
普通肝素通过不同信号通路调节 THP-1 细胞中脂多糖诱导的细胞因子产生
DOI：
10.1089/jir.2018.0042
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发表时间：
2018-07-01
影响因子：
2.3
通讯作者：
Ma, Xiaochun
中科院分区：
文献类型：
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作者：
Li, Xu;Zhao, Enfang;Ma, Xiaochun
脓毒症是由宿主对感染的先天性反应引起的复杂综合征。除了众所周知的肝素抗凝作用外，它还具有各种免疫调节特性。因此，肝素似乎是脓毒症的治疗药物。我们已经证明，普通肝素（UFH）可以抑制脂多糖（LPS）诱导的内皮细胞炎症反应。单核细胞/巨噬细胞系统是宿主免疫和抗感染免疫监视的主要贡献者。本研究的目的是确定UFH对LPS诱导的THP-1单核细胞产生细胞因子的抑制作用，并确定可能的信号通路。用UFH（0.1- 10 U/mL）处理THP-1细胞15 min，然后暴露于LPS（100 ng/mL）。免疫印迹法检测核因子B（NF-B）、磷酸化抑制因子B-（IB-）、c-Jun、c-fos、ERK 1/2、JNK和p38丝裂原活化蛋白激酶（MAPK）的表达水平。6 h后，采用酶联免疫吸附法测定IL-1、TNF-α、IL-6、IL-8和IL-18蛋白浓度。通过甲基噻唑基四唑（MTT）测定法测定细胞活力。UFH抑制LPS诱导的IB-、c-Jun、ERK 1/2、JNK和p38 MAPK的磷酸化，但不抑制c-fos。UFH还抑制LPS诱导的NF-B核转位。正如预期的那样，UFH降低LPS诱导的IL-1，TNF-α，IL-6，IL-8和IL-18蛋白水平，表明UFH通过阻断MAPK，NF-B和c-Jun信号通路对THP-1细胞具有抗炎作用。UFH可能有助于脓毒症的治疗。
Sepsis is a complex syndrome resulting from the innate host response to infection. Apart from the well-known anticoagulant effects of heparin, it also possesses various immunomodulatory properties. Thus heparin seems to be a therapeutic drug in sepsis. We have demonstrated that unfractionated heparin (UFH) can inhibit lipopolysaccharide (LPS)-induced inflammatory responses in endothelial cells. The monocyte/macrophage system is a major contributor to host immunity and immune surveillance against infection. The aim of the study is to determine the inhibitory effect of UFH on cytokine production in THP-1 monocytes induced by LPS and to define the possible signaling pathways. The THP-1 cells were treated with UFH (0.1-10U/mL) for 15min before exposure to LPS (100ng/mL). After 1h, nuclear factor-B (NF-B) and phosphorylated inhibitor B- (IB-), c-Jun, c-fos, ERK1/2, JNK, and p38 mitogen-activated protein kinase (MAPK) expression levels were evaluated by Western blot. After 6h, interleukin (IL)-1, tumor necrosis factor- (TNF-), IL-6, IL-8, and IL-18 protein concentrations were measured by enzyme-linked immunosorbent assay. Cell viability was determined by methyl thiazoyltetrazolium (MTT) assay. UFH inhibited LPS-induced phosphorylation of IB-, c-Jun, ERK1/2, JNK, and p38 MAPK but not c-fos. UFH also suppressed LPS-induced nuclear translocation of NF-B. As expected, UFH decreased LPS-induced IL-1, TNF-, IL-6, IL-8, and IL-18 protein levels, suggesting that UFH has an anti-inflammatory effect on THP-1 cells by interrupting the MAPK, NF-B, and c-Jun signaling pathways. UFH could potentially contribute to treatments for sepsis.