目的:油菜素(BR)的作用及其分子机制,来自十字花科蔬菜的吲哚植物抗毒素,在这项研究中,研究了单核细胞到巨噬细胞的分化和炎症反应。
方法:脂多糖(1µg/ml)刺激RAW264.7细胞和THP-1的炎症反应,佛波醇肉豆蔻酸盐乙酸盐(50ng/ml)诱导THP-1的单核细胞向巨噬细胞的分化。通过ELISA测定炎症介质的产生,蛋白质印迹或实时PCR。通过DCFH-DA测定法检查活性氧。
结果:50μm的油菜素抑制了脂多糖诱导的一氧化氮合酶的产生,环氧合酶-2,前列腺素E2和活性氧的90%,69%,52%和41%,分别,在RAW264.7细胞。在THP-1细胞中,BR通过抑制分化分子β和CD36簇抑制佛波醇肉豆蔻酸酯诱导的单核细胞向巨噬细胞的分化。此外,BR抑制活化B细胞(NF-κB)核因子κ轻链增强子向细胞核的转运。然而,BR激活了核因子红系衍生的2样2(Nrf2)及其靶分子血红素加氧酶-1(HO-1)和NAD(P)H:醌氧化还原酶1(NQO1),随着Nrf2核易位的增加。
结论:Brassinin通过差异调节Nrf2和NF-κB信号传导抑制单核细胞-巨噬细胞分化和炎症反应。
OBJECTIVE: The effects and molecular mechanisms of brassinin (BR), an indole phytoalexin from cruciferous vegetables, on monocyte-to-macrophage differentiation and inflammatory responses were investigated in this study.
METHODS: Inflammatory responses from RAW264.7 cells and THP-1 were stimulated by lipopolysaccharide (1 µg/ml), and monocyte-to-macrophage differentiation of THP-1 was induced by phorbol myristate acetate (50 ng/ml). The production of inflammatory mediators was determined by ELISA, Western blot or real-time PCR. Reactive oxygen species were examined by DCFH-DA assay.
RESULTS: Brassinin at 50 µm suppressed lipopolysaccharide-induced production of nitric oxide synthase, cyclooxygenase-2, prostaglandin E2 and reactive oxygen species by 90%, 69%, 52% and 41%, respectively, in RAW264.7 cells. In THP-1 cells, BR inhibited phorbol myristate acetate-induced monocyte-to-macrophage differentiation by suppressing cluster of differentiation molecule β and CD36. In addition, BR suppressed translocation of nuclear factor \'kappa-light-chain-enhancer\' of activated B cells (NF-κB) into the nucleus. However, BR activated the nuclear factor erythroid-derived 2-like 2 (Nrf2) and its target molecules hemoxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1), with an increase in nuclear translocation of Nrf2.
CONCLUSIONS: Brassinin suppressed monocyte-to-macrophage differentiation and inflammatory responses by differentially regulating Nrf2 and NF-κB signallings.