番茄红素抑制香烟烟雾提取物刺激巨噬细胞所致的炎症反应

Lycopene Inhibits Inflammatory Response of Macrophages Activated by Cigarette Smoke Extract

【目的】探讨番茄红素对香烟烟雾提取物(CSE)所诱导的人单核细胞株(THP-1)巨噬细胞炎症因子生成的影响及其作用的分子机制。【方法】用100μg/L佛波酯(PMA)诱导THP-1细胞48 h,使其分化为巨噬细胞后,分为空白组、CSE组和CSE+不同浓度番茄红素组(0.5、1和2μmol/L)。采用ELISA法检测THP-1巨噬细胞培养基中白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的含量,荧光探针DCFH-DA测定细胞内活性氧(ROS)生成水平,Western-blotting检测核p65的蛋白表达量。【结果】CSE诱导THP-1巨噬细胞后,IL-6和TNF-α合成释放水平升高,番茄红素可降低CSE诱导引起IL-6和TNF-α生成水平升高。CSE刺激后细胞内ROS生成及核p65蛋白表达增加,核因子-κB(NF-κB)活化,番茄红素可拮抗以上作用,阻止NF-κB活化。【结论】番茄红素能够有效抑制CSE诱导THP-1巨噬细胞炎症反应,该作用主要通过阻断CSE诱导细胞内ROS生成进而抑制NF-κB依赖的炎症因子IL-6和TNF-α分泌发挥的,这可能是番茄红素抑制吸烟导致的炎症反应的作用机制之一。

【Objective】To investigate the effects of lycopene on the production of proinflammatory cytokines in macro?phages of human monocyte line(THP-1)induced by cigarette smoke extract(CSE)and its possible molecular mecha?nisms.【Methods】100μg/L phorbol ester(PMA)was used to induce THP-1 cells for 48 hours,after the differentiation of THP-1 cells,which became macrophage,and cells were divided into control group,CSE group and CSE combined with different concentration(0.5,1,2μmol/L)of lycopene.The levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)were assayed by ELISA.The level of reactive oxygen species(ROS)was detected by the methods of DCFH-DA.The level of nuclear factor-κB(NF-κB)P65 protein was analyzed by Western blotting.【Results】The levels of IL-6 and TNF-αwere increased in the THP-1 macrophages after stimulated by CSE,lycopene could reduce the elevated levels of IL-6 and TNF-α.The production of ROS and the expression of nuclear P65 significantly increased after stimulated by CSE and NF-κB was activated.Lycopene could reverse the above changes and prevent the activation of NF-κB.【Conclusion】Lycopene could inhibit CSE-induced inflammatory response in THP-1 macrophages through reducing CSE-stimulated ROS production and then inhibiting the NF-κB activation and the related proinflammatory cytokines production,which is one of the mechanisms responsible for preventing smoking induced-inflammation by lycopene.