Oxidative damage to DNA induced by areca nut extract
Oxidative damage to DNA induced by areca nut extract
复制标题
DOI：
10.1016/s0165-1218(96)90018-x
复制
发表时间：
1996-01-01
期刊：
影响因子：
--
通讯作者：
Chi, CW
中科院分区：
文献类型：
--
作者：
Liu, TY;Chen, CL;Chi, CW
在台湾，人们咀嚼槟榔液，其中含有带壳的槟榔果。在其他国家，人们更喜欢成熟和干燥的无壳槟榔。在本研究中，我们比较了鲜槟榔提取物（ANE）和熟槟榔提取物（ANE）对活性氧诱导的分离DNA和CHO-K1细胞的氧化DNA损伤。这两种ANE制剂与分离的DNA在碱性环境中以剂量依赖的方式产生8-羟基-2'-脱氧鸟苷（8-OH-dG）。成熟ANE比嫩ANE产生更高水平的8-OH-dG。添加铁（II） （100 μ M）时，与1 mg/ml嫩和成熟的ANE孵育，8-OH-dG分别增加1.4倍和3.1倍。在测试ANE对细胞DNA的影响时，CHO-KI细胞被用于其对活性氧的敏感性。在CHO-K1细胞中，经过18小时的孵育，成熟的ANE比嫩的ANE更具有细胞毒性。在萌芽（r = 0.97）和成熟（r = 0.91）的ANE处理下，CHO-K1细胞的细胞毒性与8-OH-dG的形成呈正相关。在成熟ANE暴露前，细胞中添加铁螯合剂邻菲罗啉（10和20 μ M）显著提高CHO-K1细胞的存活率（p < 0.05）。此外，成熟的AWE诱导了二氯荧光素介导的荧光，表明CHO-K1细胞中过氧化氢的形成。综上所述，本研究表明，awe诱导的分离DNA和细胞DNA的氧化损伤可能是由过氧化氢的生成引起的，铁可能在这一过程中起到催化剂的作用。此外，成熟的ANE比嫩的ANE产生更高的氧化DNA损伤水平。
In Taiwan, people chew betel quid which contains tender areca nut with husk. In other countries, people prefer ripe and dried areca nut without husk. In this study, we compared the reactive oxygen species-induced oxidative DNA damage in isolated DNA and CHO-K1 cells between treatments with tender areca nut extract (ANE) and ripe ANE. Incubation of these two ANE preparations with isolated DNA generated 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in an alkaline environment in a dose-dependent manner. Ripe ANE generated higher levels of 8-OH-dG compared to tender ANE. The addition of iron(II) (100 mu M) resulted in 1.4- and 3.1-fold increases of 8-OH-dG when incubated with 1 mg/ml each of tender and ripe ANE. In testing the effect of ANE to cellular DNA, CHO-KI cells were used for its documented sensitivity to reactive oxygen species. In CHO-K1 cells, ripe ANE was more cytotoxic than tender ANE following an 18-h incubation. The cytotoxicity to CHO-K1 cells was positively correlated with the formation of 8-OH-dG following tender (r = 0.97) and ripe (r = 0.91) ANE treatment. Addition of the iron chelating agent o-phenanthroline (10 and 20 mu M) to cells prior to ripe ANE exposure significantly increased (p < 0.05) the survival of CHO-K1 cells. In addition, ripe AWE induced dichlorofluorescein-mediated fluorescence which indicated the formation of hydrogen peroxide in CHO-K1 cells. In conclusion, this study demonstrated that AWE-induced oxidative damage to isolated and cellular DNA which may result from the generation of hydrogen peroxide, and iron may serve as a catalyst in this process. Furthermore, ripe ANE generated higher oxidative DNA damage levels compared to tender ANE.