Induction of cytotoxicity and apoptosis and inhibition of cyclooxygenase-2 gene expression by eugenol-related compounds.
Induction of cytotoxicity and apoptosis and inhibition of cyclooxygenase-2 gene expression by eugenol-related compounds.
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发表时间：
2005-09
影响因子：
2
通讯作者：
N. Okada;A. Hirata;Y. Murakami;Masao Shoji;H. Sakagami;S. Fujisawa
中科院分区：
文献类型：
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作者：
N. Okada;A. Hirata;Y. Murakami;Masao Shoji;H. Sakagami;S. Fujisawa
研究了4-烯丙基-2-甲氧基苯酚（EUG）、2-甲氧基-4-甲基苯酚（MMP）、3,3'-二甲氧基-5,5'-二-2-丙基-1,1'-联苯-2,2'-二醇（双-EUG）和3,3'-二-甲氧基-5,5'-二甲基-1,1'-联苯-2,2'-二醇（双-MMP）对HL-60白血病细胞的细胞毒性和核体间DNA断裂的诱导作用。EUG、MMP、bis-EUG和bis-MMP的50%细胞毒浓度（CC50）分别为0.38 mM、0.38 mM、0.18 mM和0.20 mM。bis-EUG诱导DNA断裂最强烈，其次是EUG、MMP和bis-MMP。经丙烯酰胺凝胶电泳检测，MnSOD的表达和Cu/ZnSOD活性均被EUG抑制，提示线粒体功能障碍。RT-PCR检测发现，1 mM EUG作用1小时后，HL-60细胞中MnSOD和Cu/ZnSOD mrna的表达明显受到抑制。此外，添加5 mM n -乙酰半胱氨酸（NAC）或谷胱甘肽（GSH）可以增强EUG对SOD mrna表达的抑制作用，而单独添加NAC或GSH则不会影响SOD mrna的表达。高浓度NAC或GSH显著增强EUG的细胞毒性，这可能与EUG与GSH或NAC协同作用抑制SOD mrna表达有关。采用Northern blot方法研究了丁香酚相关化合物对脂多糖（LPS）诱导的环氧化酶-2 （COX-2）基因表达的调控作用。Bis-EUG、MMP和bis-MMP分别在300、500和500 μ m浓度下抑制COX-2基因表达。相比之下，EUG在10-500微米的大浓度范围内没有发现抑制作用，这可能是由于该化合物具有强大的促氧化活性，导致线粒体功能障碍。丁香酚相关化合物，特别是双丁香酚，可以作为非甾体抗炎药（NSAID）样化合物。
Induction of cytotoxicity and internucleosomal DNA fragmentation by 4-allyl-2-methoxyphenol (eugenol, EUG), 2-methoxy-4-methylphenol (MMP), 3,3'-dimethoxy-5,5'-di-2-propenyl-1,1'-biphenyl-2,2'-diol (bis-EUG) and 3,3'-di-methoxy-5,5'-dimethyl-1,1'-biphenyl-2,2'-diol (bis-MMP) were investigated in HL-60 leukemia cells. The 50% cytotoxic concentrations (CC50) for EUG, MMP, bis-EUG and bis-MMP were 0.38 mM, 0.38 mM, 0.18 mM and 0.20 mM, respectively. DNA fragmentation was induced most strongly by bis-EUG, followed by EUG, MMP and bis-MMP. The expression of MnSOD and, less strongly, Cu/ZnSOD activity, as assessed by acrylamide gel electrophoresis, was inhibited by EUG, suggesting mitochondrial dysfunction. The expression of the mRNAs for MnSOD and Cu/ZnSOD in HL-60 cells, as assessed by RT-PCR, was significantly inhibited by treatment with 1 mM EUG for 1 hour. Furthermore, inhibition of SOD mRNAs expression by EUG was strongly potentiated by the addition of 5 mM N-acetyl cysteine (NAC) or glutathione (GSH), whereas NAC or GSH alone did not affect the expression of SOD mRNAs. The cytotoxicity of EUG was significantly enhanced by high concentrations of NAC or GSH, which may be attributed to the inhibition of SOD mRNAs expression by the synergistic action of EUG and GSH or NAC. The regulatory effects of eugenol-related compounds on lipopolysaccharide (LPS)-induced cyclooxygenase-2 (COX-2) gene expression in RAW 264.7 cells were investigated by Northern blot analysis. Bis-EUG, MMP and bis-MMP inhibited COX-2 gene expression at concentrations of 300 microM, 500 microM and 500 microM, respectively. In contrast, no inhibitory effect of EUG was found over the wide concentration range of 10-500 microM, possibly as a result of the extensive mitochondrial dysfunction induced by this compound, which possesses potent pro-oxidative activity. Eugenol-related compounds, particularly bis-EUG, may act as nonsteroidal anti-inflammatory drug (NSAID)-like compounds.