Radical Production and Apoptosis of Eugenol-Related Compounds

丁子香酚相关化合物的自由基产生和细胞凋亡

基本信息

批准号：
13672119
负责人：
OKADA Norihisa
金额：
$ 1.6万
依托单位：
Meikai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672119/
关键词：
Methoxyphenol Eugenol Radical-induced cytotoxicity Apoptosis McSOD Cu ZnSOD Catalytic effect Caspase Mn-SOD CU Zn-SOD

项目摘要

We investigated synthesized eugenol related compound-induced cytotoxicity and apoptosis against human cancer cells (HSC-2, HSC-3, HSG and HL-60 cells). 2-methoxy-4-allylphenol (eugenol, EUG), 2-methoxy-4-methylphenol (MMP) and their dimers (bis-EUG, bis-MMP) showed the increasing cytotoxicity with increasing hydrophobicity (log P). Whereas the cytotoxicity of 2-t-butyl-4-metylphenol and its dimers (bis-BHA) and 2, 6-t-butyl-4-methoxyphenol were decreased by increasing hydrophobicity. These findings might be connected with the chemical structure of bulky functional substitutes of 2-t-butyl group. HL-60 cells alone underwent DNA fragmentation and the degree of fragmentation for BHA was greater than that for EUG. In dimers, bis-EUG caused DNA fragmentation at a lower concentration and its Cu/ZnSOD activity in HL-60 cells was significantly larger than MnSOD counterpart. Next, we examined the appearance of mRNA in SOD elements during an apoptosis process using RT-PCR method. MnSOD and Cu/ZnSOD mRNA were found when the concentration of EUG was two-fold greater than that of CC50, possibly due to apoptosis. Also, we examined the cytotoxicity of EUG in the presence of antioxidants with SH-groups such as glutathione, glutathione ethyl ester and cysteine. Antioxidant with SH-groups enhanced the cytotoxicity of EUG, suggesting that EUG acts as a prooxidant. Caspase activations of HL-60 cells induced by EUG with the antioxidants were investigated, resulting in that it activated both exogenous and endogenous paths, and consequently activated caspase-3. However, during activation, EUG having antioxidants with SH-groups may induce not only apoptosis but also necrosis.

我们研究了对人类癌细胞的合成相关化合物诱导的细胞毒性和凋亡（HSC-2，HSC-3，HSG和HL-60细胞）。 2-甲氧基-4-甲苯酚（Eugenol，EUG），2-甲氧基-4-甲基苯酚（MMP）及其二聚体（BIS-EUG，BIS-MMP）显示出随着疏水性增加而增加的细胞毒性率（log p）。而2-T-叔丁基-4-甲醇及其二聚体（BIS-BHA）和2，6-T-叔丁基-4-甲氧基苯酚的细胞毒性通过增加而降低。这些发现可能与2-T-丁基基团的笨重功能替代物的化学结构有关。单独的HL-60细胞经历了DNA片段化，而BHA的片段化程度大于EUG的片段化程度。在二聚体中，BIS-EUG在较低的浓度下引起DNA片段化，其HL-60细胞中其Cu/ZnSOD活性明显大于MNSOD对应物。接下来，我们使用RT-PCR方法在凋亡过程中检查了SOD元素中mRNA的出现。当EUG的浓度大于CC50时，发现MNSOD和Cu/Znsod mRNA，可能是由于凋亡所致。此外，我们在存在抗氧化剂的情况下检查了EUG的细胞毒性，例如谷胱甘肽，谷胱甘肽乙基酯和半胱氨酸。用SH组的抗氧化剂增强了EUG的细胞毒性，表明EUG是一种促氧化剂。研究了用抗氧化剂诱导的HL-60细胞的caspase激活，从而使其激活了外源性和内源性路径，因此激活了Caspase-3。但是，在激活过程中，具有抗sh组的抗氧化剂的EUG不仅会诱导凋亡，而且会诱发坏死。