Comparative inhibitory effects of magnolol, honokiol, eugenol and bis-eugenol on cyclooxygenase-2 expression and nuclear factor-kappa B activation in RAW264.7 macrophage-like cells stimulated with fimbriae of Porphyromonas gingivalis.
Comparative inhibitory effects of magnolol, honokiol, eugenol and bis-eugenol on cyclooxygenase-2 expression and nuclear factor-kappa B activation in RAW264.7 macrophage-like cells stimulated with fimbriae of Porphyromonas gingivalis.
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发表时间：
2012-11
期刊：
影响因子：
2.3
通讯作者：
Y. Murakami;Akifumi Kawata;Yuya Seki;Teho Koh;Kenji Yuhara;Takehisa Maruyama;M. Machino;S. Ito;Y. Kadoma;S. Fujisawa
中科院分区：
文献类型：
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作者：
Y. Murakami;Akifumi Kawata;Yuya Seki;Teho Koh;Kenji Yuhara;Takehisa Maruyama;M. Machino;S. Ito;Y. Kadoma;S. Fujisawa
背景厚朴酚及其相关化合物的抗炎活性是目前关注的焦点。在本研究中，这些化合物对环氧化酶（考克斯-2）表达和核因子-κ B（NF-κB B）激活的抑制作用在口腔厌氧菌牙龈卟啉单胞菌菌毛刺激的RAW 264. 7巨噬细胞样细胞中进行了研究。材料与方法采用CCK-8细胞计数试剂盒测定厚朴酚、和厚朴酚、丁香酚和双丁香酚对RAW264.7细胞的细胞毒性。通过实时聚合酶链反应（PCR）研究了这些化合物对暴露于菌毛刺激的考克斯-2 mRNA表达的调节作用。采用酶联免疫吸附试验（ELISA）-样微孔比色转录因子活性测定（Trans-AM）和western blot分析检测NF-κB活化。在近厌氧条件下，采用诱导期法测定了甲基丙烯酸甲酯-偶氮二异丁腈（AIBN）聚合体系的自由基清除活性。在密度泛函理论（DFT）B3 LYP/6- 31 G * 水平上计算了酚键离解焓（BDE）和轨道能.结果：对RAW264.7细胞的细胞毒作用顺序为双丁香酚>丁香酚>和厚朴酚>厚朴酚，对自由基的清除作用顺序为和厚朴酚>双丁香酚>厚朴酚>丁香酚。厚朴酚和和厚朴酚在非细胞毒性浓度下显著抑制菌毛诱导的考克斯-2表达。木兰酚和和厚朴酚显著抑制菌毛刺激的NF-κB与其共有序列的结合以及抑制剂κB-α的磷酸化依赖性蛋白水解，而丁香酚和双丁香酚不抑制考克斯-2表达和NF-κB活化。厚朴酚和和厚朴酚具有较高的电负性（χ）值。结论厚朴酚、和厚朴酚具有抗氧化、低细胞毒性和抗炎活性。这些化合物可能能够预防由口腔细菌引起的慢性炎性疾病。
BACKGROUND The anti-inflammatory activity of magnolol and related compounds is currently a focus of interest. In the present study, the inhibitory effects of these compounds on cyclooxygenase (COX-2) expression and nuclear factor-kappa B (NF-κB) activation were investigated in RAW264.7 macrophage-like cells stimulated with the fimbriae of Porphyromonas gingivalis, an oral anaerobe. MATERIALS AND METHODS The cytotoxicity of magnolol, honokiol, eugenol and bis-eugenol against RAW264.7 cells was determined using a cell counting kit (CCK-8). The regulatory effect of these compounds on the expression of COX-2 mRNA, stimulated by exposure to the fimbriae was investigated by real-time polymerase chain reaction (PCR). NF-κB activation was evaluated by enzyme-linked immunosorbent assay (ELISA)-like microwell colorimetric transcription factor activity assay (Trans-AM) and western blot analysis. The radical-scavenging activity was determined using the induction period method in the methyl methacrylate-azobisisobutyronitrile (AIBN) polymerization system under nearly anaerobic conditions. The phenolic bond dissociation enthalpy (BDE) and orbital energy were calculated at the density functional theory (DFT) B3LYP/6-31G* level. RESULTS The cytotoxicity against RAW264.7 cells declined in the order bis-eugenol>eugenol> honokiol>magnolol, whereas the radical-scavenging activity declined in the order honokiol, bis-eugenol>magnolol> eugenol. Magnolol and honokiol significantly inhibited the fimbria-induced expression of COX-2 at non-cytotoxic concentrations. Both the fimbria-stimulated binding of NF-κB to its consensus sequence and phosphorylation-dependent proteolysis of inhibitor κB-α were markedly inhibited by magnilol and honokiol, whereas eugenol and bis-eugenol did not inhibit COX-2 expression and NF-κB activation. Magnolol and honokiol possessed a high electronegativity (χ) value. CONCLUSION Magnolol and honokiol exhibit antioxidative activity, low cytotoxicity, and anti-inflammatory activity. These compounds may be capable of preventing chronic inflammatory diseases induced by oral bacteria.