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Effects of hypertonic saline on macrophage migration inhibitory factor in traumatic conditions

机译:高渗盐水对创伤条件下巨噬细胞迁移抑制因子的影响

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摘要

Trauma-induced suppression of cellular immune function contributes to sepsis, multiple organ dysfunction syndrome (MODS) and mortality. Macrophage migration inhibitory factor (MIF) has been revealed to be central to several immune responses. However, the role of MIF in trauma-like conditions is unknown. Therefore, the present study evaluated MIF in macrophages and polymorphonuclear neutrophils (PMNs). The effects of hypertonic saline (HTS) on lipopolysaccharide (LPS)-induced MIF levels were evaluated in macrophages. MIF concentrations were determined by an enzyme-linked immnosorbent assay (ELISA) and cell lysates were used for western blot analysis. The effects of HTS on N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced MIF were evaluated in PMNs. MIF concentrations were determined by ELISA, western blotting and real time-polymerase chain reaction (RT-PCR) to determine MIF expression. MIF levels, which were measured by the ELISA, increased by 1.24 +/- 0.38 ng/ml in the supernatants of LPS-stimulated macrophages compared with the controls (0.79 +/- 0.07 ng/ml) at 2 h. HTS 10 (150 mmol/l) partially restored MIF levels (0.84 +/- 0.22 ng/ml; P<0.05). Also, western blotting was performed and MIF protein levels were higher in the LPS-stimulated macrphages (20% increase in band density) compared with the controls (P<0.05). The addition of HTS decreased MIF protein expression. MIF levels in fMLP-stimulated PMN cells were unchanged compared with the controls according to the ELISA, western blotting and RT-PCR. No effects were observed following treatment with HTS. MIF concentrations and MIF expression were higher in LPS-stimulated macrophages than controls and HTS restored M IF levels to those of the controls. M IF levels were unchanged in PMNs stimulated by fMLP.
机译:创伤引起的细胞免疫功能抑制导致败血症,多器官功能障碍综合症(MODS)和死亡率。巨噬细胞迁移抑制因子(MIF)已被证明是几种免疫反应的核心。但是,MIF在类似创伤的疾病中的作用尚不清楚。因此,本研究评估了巨噬细胞和多形核中性粒细胞(PMNs)中的MIF。在巨噬细胞中评估了高渗盐水(HTS)对脂多糖(LPS)诱导的MIF水平的影响。 MIF浓度通过酶联免疫吸附测定(ELISA)确定,细胞裂解液用于Western印迹分析。在PMN中评估了HTS对N-甲酰基-甲硫基-亮氨酰-苯丙氨酸(fMLP)诱导的MIF的影响。 MIF浓度通过ELISA,蛋白质印迹和实时聚合酶链反应(RT-PCR)确定MIF表达。通过ELISA测定的MIF水平在2 h时与LPS刺激的巨噬细胞上清液相比,在对照中(0.79 +/- 0.07 ng / ml)增加了1.24 +/- 0.38 ng / ml。 HTS 10(150 mmol / l)部分恢复了MIF水平(0.84 +/- 0.22 ng / ml; P <0.05)。另外,与对照相比,在LPS刺激的巨噬细胞中进行了蛋白质印迹并且MIF蛋白水平更高(条带密度增加了20%)(P <0.05)。 HTS的添加降低了MIF蛋白的表达。根据ELISA,蛋白质印迹和RT-PCR,与对照相比,fMLP刺激的PMN细胞中的MIF水平没有变化。用HTS治疗后未观察到效果。 LPS刺激的巨噬细胞中的MIF浓度和MIF表达高于对照组,而HTS恢复的M IF水平高于对照组。由fMLP刺激的PMN中的M IF水平没有变化。

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