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Objectives The purpose of this study is to investigate anti-inflammatory effects of toluhydroquinone in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods Toluhydroquinone was purified from a fungal strain, Aspergillus sp. We investigated that levels of nitric oxide (NO) using Griess reagent, production of prostaglandin E2 (PGE2) and pro-inflammatory cytokines using ELISA assay. We conducted Western blot analysis to investigate regulatory effects of toluhydroquinone on expression of inducible nitric oxide synthase (iNOS), cyclooxyganse-2 (COX-2), nuclear factor-κB (NF-κB), Akt and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW264.7 cells. The translocation of NF-κB was detected by immunofluorescence staining. Key findings Toluhydroquinone inhibited production of NO and PGE2 via suppressing protein expression of iNOS and COX-2, respectively. Secretion and expression of inflammatory cytokines were down-regulated by toluhydroquinone as well. Toluhydroquinone reduced phosphorylation of Akt, NF-κB and MAPKs. Moreover, toluhydroquinone inhibited translocation of NF-κB from the cytosol into the nucleus. Conclusions We revealed that inhibitory effects of toluhydroquinone on expression of inflammatory mediators are induced through inactivation of Akt, NF-κB and MAPKs. Thus, our results suggest that toluhydroquinone may be used for a potential anti-inflammatory reagent. 翻译.PNG |
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目的 本文旨在研究甲基氢醌在脂多糖(LPS)激活的RAW264.7细胞中的抗炎效果。 方法 甲基氢醌是由真菌菌株,曲霉菌属纯化而得到的。我们使用格里斯试剂检测了一氧化氮(NO)水平,采用酶联免疫吸附测定法检测了前列腺素E2(PGE2)和促炎细胞因子的产生。我们进行了免疫印迹分析以检测甲基氢醌对脂多糖激活RAW264.7细胞中的诱导型一氧化氮合酶(iNOS)、环氧合酶-2 (COX - 2)、核因子κB(NF - κB)、蛋白激酶和丝裂原活化蛋白激酶(MAPKs)表达的调节作用。通过免疫荧光染色法检测核因子κB的易位。 主要发现 甲基氢醌分别通过抑制诱导型一氧化氮合酶和环氧合酶-2的蛋白表达阻碍了NO和前列腺素E2的产生。同时甲基氢醌也下调了炎性细胞因子的分泌和表达。甲基氢醌减少了蛋白激酶、核因子κB和原活化蛋白激酶的磷酰化。而且，甲基氢醌阻止了核因子κB从细胞液向细胞核的易位。 结论 我们发现甲基氢醌对炎性介质表达的抑制作用是通过蛋白激酶、核因子κB和原活化蛋白激酶的失活而激发的。综上所述,我们的研究结果表明甲基氢醌可以用于潜在的抗炎试剂。 |
2楼2017-05-24 10:16:07