Oxidative Stress Induces Expression of the Toll-Like Receptors (TLRs) 2 and 4 in the Human Peripheral Blood Mononuclear Cells: Implications for Metabolic Inflammation.
Oxidative Stress Induces Expression of the Toll-Like Receptors (TLRs) 2 and 4 in the Human Peripheral Blood Mononuclear Cells: Implications for Metabolic Inflammation.
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DOI：
10.33594/000000117
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发表时间：
2019-01-01
期刊：
影响因子：
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通讯作者：
Sindhu, Sardar
中科院分区：
文献类型：
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作者：
Akhter, Nadeem;Madhoun, Ashraf;Sindhu, Sardar
背景/目的：先天免疫toll样受体（TLRs）正逐渐成为营养传感器。肥胖中脂肪组织的氧化应激是病理生理改变的关键早期触发因素。TLR2/TLR4脂肪上调与人类胰岛素抵抗有关；然而，氧化应激是否可以调节免疫细胞中TLR2/4及相关免疫代谢调节因子（IRF3/5）的表达尚不清楚。因此，我们在氧化应激诱导下评估了它们与促炎细胞因子在人PBMC中的表达。方法：采用Ficoll-Paque法从健康献血者血液中分离PBMC，用H2O2诱导细胞氧化应激。DCFH-DA法测定ROS。目的基因和蛋白表达分别采用实时RT-PCR和流式细胞术/共聚焦显微镜检测。在ros抑制剂或瘦素/脂多糖/脂肪酸存在的情况下，H2O2对TLR2/4的表达也进行了评估。western blotting检测磷酸化/总ERK1/2、c-Jun、p38和NF-kappaB的表达。数据（均数±SEM）采用未配对学生t检验或方差分析进行比较；所有的假定值
BACKGROUND/AIMS: Innate immune toll-like receptors (TLRs) are emerging as nutrient sensors. Oxidative stress in the adipose tissue in obesity acts as a critical early trigger of altered pathophysiology. TLR2/TLR4 adipose upregulation has been associated with insulin resistance in humans; however, it remains unclear whether oxidative stress can modulate expression of TLR2/4 and related immune-metabolic regulators (IRF3/5) in immune cells. We, therefore, assessed their expression along with proinflammatory cytokines in the human PBMC following induction of oxidative stress.METHODS: PBMC were isolated from blood of healthy donors using Ficoll-Paque method and cells were treated with H2O2 to induce oxidative stress. ROS was measured by DCFH-DA assay. Target gene and protein expression was determined using real-time RT-PCR and flow cytometry/confocal microscopy, respectively. TLR2/4 expression by H2O2 in presence of ROS-inhibitors or leptin/LPS/fatty acids was also assessed. Expression of phosphorylated/total ERK1/2, c-Jun, p38, and NF-kappaB was determined by western blotting. The data (mean±SEM) were compared using unpaired student's t-test or ANOVA; all P-values