粉防己碱对脂多糖诱导下RAW264.7细胞COX-2/PGE_2、iNOS/NO表达的影响

Effects of tetrandrine on COX-2/PGE_2,iNOS/NO expression in LPS-stimulated RAW264.7 cells

本实验旨在观察粉防己碱(Tet)对脂多糖(LPS)诱导下RAW264.7细胞炎症模型的抗炎作用。采用LPS1μg/mL刺激生长良好的RAW264.7巨噬细胞建立细胞炎症模型。在不同浓度Tet(1×10-8,1×10-7,1×10-6 mol/L)作用下,用Western blotting检测各组细胞中环加氧酶-2(COX-2)和诱导性一氧化氮激酶(iNOS)的表达,用NO检测试剂盒检测细胞培养液中NO的含量,酶免疫分析法检测培养液中前列腺素E2(PGE2)含量。结果显示Tet剂量依赖性地抑制LPS诱导的RAW264.7细胞PGE2、NO的表达,同时抑制其合成酶COX-2和iNOS的表达。表明Tet具有抑制LPS诱导的RAW264.7细胞炎症反应的作用,其抗炎机制可能通过抑制COX-2、iNOS的表达,从而抑制其下游炎性介质NO、PGE2的表达有关。

The effects of tetrandrine（Tet） on the expression of cyclooxygenase-2（COX-2）,prostaglandin E2（PGE2）,inducible isoforms of nitric oxide synthetase（iNOS） and nitric oxide（NO） in LPS-stimulated RAW264.7 cells were investigated.RAW 264.7 cells were pretreated with LPS 1 μg/mL to set up the cell inflammatory model;various concentrations（1×10-8,1×10-7,1×10-6 mol/L） of Tet were administered to explore its effect on the expression of COX-2/PGE2,iNOS/NO in these LPS-stimulated cells.Western blotting was applied to detect the expression of intracellular COX-2 and iNOS;special NO kit was used to detect the level of NO,and enzyme immunoassay（EIA） kit to that of PGE2.The results showed that Tet significantly decreased the expression of PGE2 and NO as well as COX-2 and iNOS synthetase in LPS-stimulated RAW264.7 cells in a dose-dependent manner.So it suggests that Tet can inhibit the inflammation response in LPS-stimulated RAW264.7 cell,which might be mediated by down-regulating the expression of PGE2 and NO through the inhibition of COX-2 and iNOS synthetase.