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(50 ng/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。
Objective To 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(50 ng/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.Results R406 inhibited the mRNA expression of TNF-α,IL-1βand 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.Conclusion R406 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.
作者
林杰
张波
LIN Jie;ZHANG Bo(Dept.of Medical Ultrasound,Shanghai East Hospital,School of Medicine,Tongji University,Shanghai 200120,China)
出处
《同济大学学报（医学版）》
2021年第4期467-471,共5页
Journal of Tongji University(Medical Science)
基金
国家自然科学基金(81871361)。
