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  • 林杰,张波.R406抑制脂多糖诱导的RAW264.7细胞炎症及其机制研究[J].同济大学学报(医学版),2021,42(4):467-471.    [点击复制]
  • LIN Jie,ZHANG Bo.R406 inhibits LPS-induced inflammation in RAW264.7 cells and its mechanism[J].同济大学学报(医学版),2021,42(4):467-471.   [点击复制]
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R406抑制脂多糖诱导的RAW264.7细胞炎症及其机制研究
林杰,张波
0
(同济大学附属东方医院医学超声科,上海200120)
摘要:
目的研究脾酪氨酸激酶抑制剂R406对脂多糖(lipopolysaccharide, LPS)诱导的RAW264.7细胞炎症抑制作用以及其可能的机制。方法使用LPS(50ng/mL)刺激RAW264.7建立体外细胞炎症模型后与R406共孵育。通过酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)检测细胞上清液中肿瘤坏死因子(tumor necrosis factor, TNF-α)、单核细胞趋化蛋白1(C-C motif chemokine 2/human macrophage chemoattractant protein-1, MCP-1)、一氧化氮(nitric oxide, NO)的水平;通过PCR检测细胞中TNF-α、白细胞介素-1β(interleukin-1β, IL-1β)、诱导型一氧化氮合酶(inductible nitric oxide synthase, iNOs)的mRNA水平;通过Western印迹法检测核转录因子κB(NF-κB)的磷酸化水平。结果R406能抑制LPS诱导的巨噬细胞TNF-α、IL-1β、iNOs的mRNA表达,同时能抑制细胞炎症因子的释放且呈剂量相关性(P<0.05)。除此之外,R406对NF-κB的磷酸化起抑制作用。结论R406能抑制LPS诱导的巨噬细胞炎症因子的释放,其作用可能与抑制NF-κB的磷酸化信号通路相关。
关键词:  R406  脾酪氨酸激酶  单核巨噬细胞  脂多糖  炎症
DOI:10.12289/j.issn.1008-0392.21116
投稿时间:2021-03-25
基金项目:国家自然科学基金(81871361)
R406 inhibits LPS-induced inflammation in RAW264.7 cells and its mechanism
LIN Jie,ZHANG Bo
(Dept. of Medical Ultrasound, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China)
Abstract:
ObjectiveTo investigate the effect of tyrosine kinase inhibitor R406 on the inflammation of mouse macrophage RAW264.7 cells induced by lipopolysaccharide(LPS) and its mechanism. Methods Inflammation was induced by LPS(50ng/mL) in RAW246.7 cells, the cells were cocultrued with R406(0.5, 1.0, 1.5μmol/L). The levels of TNF-α, CCL-2/MCP-1, NO in the cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA); the mRNA expression levels of TNF-α, IL-1β, iNOs in the cells were detected by RT-PCR. The phosphorylation level of nuclear transcription factor NF-κB was detected by Western blot. ResultsR406 inhibited the mRNA expression of TNF-α, IL-1β and iNOs in macrophages induced by LPS, and at the same time inhibited the release of cellular inflammatory factors(P<0.05) in a dose-dependent manner. In addition, R406 inhibited the phosphorylation of NF-κB. ConclusionR406 can inhibit the release of inflammatory factors in macrophages induced by LPS and its effect may be related to the inhibition of NF-κB phosphorylation signaling pathway.
Key words:  R406  spleen tyrosine kinase  RAW264.7  lipopolysaccharide  inflammation

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