微生物与感染
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微生物与感染  2016, Vol. 11 Issue (2): 100-105    
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内毒素耐受对核苷酸结合寡聚化结构域2信号通路的影响
张辉军1,王葆青1,瞿介明2
1. 复旦大学附属中山医院肺科,上海 200032;2. 上海交通大学医学院附属瑞金医院呼吸科,上海 200025
Effect of endotoxin tolerance on nucleotide­-binding oligomerization domain containing 2 signaling pathway
ZHANG Huijun1, WANG Baoqing1, QU Jieming2
1. Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, Shanghai 200032, China; 2. Department of Pulmonary Medicine, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
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摘要: 为研究内毒素耐受对核苷酸结合寡聚化结构域2(nucleotide­-binding oligomerization domain containing 2,NOD2)信号通路的影响,将小鼠单核-­巨噬细胞RAW264.7分为两组,分别给予小剂量脂多糖(lipopolysaccharide,LPS)(100 ng/mL)或磷酸盐缓冲液(phosphate buffered saline,PBS)预处理20 h,建立内毒素耐受组和对照组。每组细胞分别给予大剂量LPS(1 000 ng/mL)或热灭活烟曲霉孢子刺激,于刺激后0、2、6、12、24 h采用定量聚合酶链反应(polymerase chain reaction,PCR)检测细胞NOD2、受体相互作用蛋白2(recepto-r­interacting protein 2,RIP2)和肿瘤坏死因子α(tumor necrosis factor α,TNF-­α)mRNA表达;采用酶联免疫吸附试验(enzyme-­linked immunosorbent assay,ELISA)检测细胞上清液中白细胞介素8(interleukin 8,IL­-8)和TNF-­α浓度。结果显示,内毒素耐受组无论是大剂量LPS还是热灭活烟曲霉孢子刺激均不能增加NOD2、RIP2 和TNF­-α mRNA表达及细胞上清液中IL-­8、TNF­-α浓度;而对照组大剂量LPS和热灭活烟曲霉孢子刺激均可提高NOD2、RIP2 和TNF-­α mRNA表达及细胞上清液中IL-­8、TNF­-α浓度,尤以刺激后12 h增加显著,与刺激前(0 h)比较,差异有统计学意义(P<0.05)。结果提示,内毒素耐受可能对NOD2信号通路有抑制作用。

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张辉军1
王葆青1
瞿介明2
关键词 内毒素耐受脂多糖核苷酸结合寡聚化结构域2烟曲霉    
Abstract:To investigate the potential effect of endotoxin tolerance on nucleotide­-binding oligomerization domain containing 2 (NOD2) signaling pathway, murine macrophage RAW264.7 cells were pretreated with lipopolysaccharide (LPS, 100 ng/mL) or phosphate buffered saline (PBS) for 20 h to establish endotoxin tolerance group and control group respectively. Both groups were subjected to the treatment with LPS (1 000 ng/mL) or heat­killed Aspergillus fumigatus (HAf). The mRNA levels of NOD2, receptor-­interacting protein 2 (RIP2) and tumor necrosis factor α (TNF-­α) were measured at 0, 2, 6, 12, 24 h after stimulation by real-­time quantitative polymerase chain reaction (PCR). The concentrations of secreted TNF-­α and interleukin 8 (IL­-8) were also quantified by enzyme-­linked immunosorbent assay (ELISA). The collective results showed that neither LPS nor HAf had any significant effect on NOD2, RIP2, and TNF­-α transcription or TNF­-α and IL­-8 secretion in the endotoxin tolerance group. On the contrary, a clear up-­regulation was detected in the control group upon either LPS or HAf treatment, concomitant with a peak at 12 h after stimulation. These data indicate that the NOD2 signaling pathway might be suppressed under endotoxin tolerance conditions.

Key wordsEndotoxin tolerance    Lipopolysaccharide    Nucleotide­binding oligomerization domain containing 2;Aspergillus fumigatus
基金资助:教育部高等学校博士学科点-新教师基金资助(20090071120029)

通讯作者: 瞿介明   
引用本文:   
张辉军1,王葆青1,瞿介明2 . 内毒素耐受对核苷酸结合寡聚化结构域2信号通路的影响[J]. 微生物与感染, 2016, 11(2): 100-105.
ZHANG Huijun1, WANG Baoqing1, QU Jieming2. Effect of endotoxin tolerance on nucleotide­-binding oligomerization domain containing 2 signaling pathway. JOURNAL OF MICROBES AND INFECTIONS, 2016, 11(2): 100-105.
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