圆齿野鸦椿果实总黄酮对LPS诱导的RAW264.7细胞炎症反应的抑制作用

The anti-inflammatory effect of total flavonoids from Euscaphis konishii Hayata pericarp (TFEP)on the LPS-induced RAW264.7 macrophages

探讨圆齿野鸦椿果实总黄酮(TFEP)对脂多糖(LPS)诱导的巨噬细胞RAW264.7细胞炎症反应的抑制效果.利用一氧化氮(NO)试剂盒检测不同浓度TFEP对LPS诱导RAW264.7细胞释放NO含量的变化;酶联免疫吸附法(ELISA)测定PGE2、白介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)分泌物的表达;实时定量逆转录聚合酶链反应法(qRT-PCR)检测诱导型一氧化氮合酶(iNOS)、TNF-α和IL-6 mRNA的表达;蛋白质免疫印迹法(western blot)检测炎症NF-κβ/Stat3信号通路相关蛋白的表达.结果表明,TFEP对细胞产生NO的半数抑制浓度(IC50)为78.47μg·mL^-1,且各试验浓度下对RAW264.7细胞增殖均无明显抑制作用,表现出低毒性和强抗炎活性.与LPS模型组相比,TFEP能显著降低LPS诱导的NO的分泌;抑制iNOS、TNF-α和IL-6 mRNA的表达,抑制炎症NF-κβ/Stat3信号通路的激活.因此,圆齿野鸦椿果实总黄酮对LPS诱导的RAW264.7细胞炎症反应有良好的抑制作用.

To investigate whether total flavonoids from Euscaphis konishii Hayata pericarp(TFEP)exerted anti-inflammatory function on LPS-induced RAW264.7 macrophages,NO released by macrophages was quantified using NO kit assay,and PGE2,IL-6 and TNF-αexcreted by inflammatory cells were detected by enzyme linked immunosorbent assay(ELISA).The mRNA expression of iNOS,TNF-αand IL-6 were evaluated using qRT-PCR,and levels of signaling proteins of NF-κβand Stat3 were quantified by Western blot.The half inhibitory concentration(IC50)of TFEP on NO production was 78.47μg􀅰mL-1,without obvious inhibitory effect on each test concentration,indicating a low toxicity and strong anti-inflammatory activity.Compared with the LPS model group,TFEP significantly reduced the NO production of LPS-induced macrophages.mRNA expressions of iNOS,TNF-αand IL-6 were significantly inhibited,so as the activation of NF-κβand Stat3 signaling pathways.In summary,TFEP can inhibit the inflammation of RAW264.7 cells induced by LPS.