癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (5): 372-376.doi: 10.3969/j.issn.1004-616x.2009.05.010

• 论著 • 上一篇    下一篇

喹乙醇诱导HepG2细胞线粒体的氧化损伤

邹家杰;张 婷;汤树生;陈 倩;靳 溪;陈开跑;肖希龙   

  1. 中国农业大学动物医学院,北京 100193
  • 收稿日期:2009-01-05 修回日期:2009-04-23 出版日期:2009-09-30 发布日期:2009-09-30
  • 通讯作者: 肖希龙

Oxidative Damage of Mitochondria by Olaquindox in HepG2 Cells

ZOU Jia-jie; ZHANG Ting; TANG Shu-sheng; CHEN Qian; JIN Xi;CHEN Kai-pao; XIAO Xi-long   

  1. Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agriculture University, Beijing 100193, China
  • Received:2009-01-05 Revised:2009-04-23 Online:2009-09-30 Published:2009-09-30
  • Contact: XIAO Xi-long

摘要: 背景与目的: 探讨喹乙醇(olaquindox)对人肝癌细胞(HepG2)线粒体的氧化损伤作用。 材料与方法: 用0、200、400和800 μg/ml喹乙醇处理HepG2细胞24 h后,采用噻唑蓝(MTT)法确定喹乙醇对HepG2细胞的IC50;分子探针2',7'-二氯荧光黄双乙酸盐(2',7'-dichlorodihydrofluorescein diacetate, DCFDA)和双氢溴乙啶(dihydroethidium, DHE)检测细胞内活性氧(reactive oxygen species, ROS)含量;罗丹明123检测线粒体膜电位(△Ψm)变化;钙离子荧光探针Fluo-3AM检测胞浆游离钙离子浓度并用定量PCR方法检测线粒体DNA(mtDNA)和核DNA(nDNA)的损伤情况。 结果: 随着喹乙醇浓度的增加,HepG2细胞的存活率逐渐降低(P均<0.01),呈时间-剂量-反应关系;不同浓度的喹乙醇作用细胞24 h后,随着喹乙醇浓度的增加细胞内ROS和胞浆游离钙离子浓度显著增加(P<0.05或P<0.01),△Ψm明显降低且呈剂量-反应关系;喹乙醇能引起HepG2细胞mtDNA和nDNA损伤(P均<0.01),且mtDNA损伤程度显著高于nDNA,并呈剂量-反应关系。 结论: 喹乙醇在体外实验中,可诱导HepG2细胞线粒体氧化性损伤。

关键词: 喹乙醇, 线粒体, 活性氧, 线粒体膜电位, 钙离子

Abstract: BACKGROUND AND AIM: To investigate oxidative damage of mitochondria induced by olaquindox in human hepatoma G2 (HepG2) cells. MATERIALS AND METHODS: HepG2 cells were treated with 0,200,400 and 800 μg/ml olaquindox for 24 h. The value of IC50 of olaquindox in HepG2 cells was determined by MTT assay. The levels of reactive oxygen species (ROS) were detected by 2, 7-dichlorodihydrofluorescein diacetate (DCFDA) and Dihydroethidium (DHE). The level of mitochondrial membrane potential(△Ψm) and calcium concentration were measured by Rhodamine123(Rh-123) and Fluo-3AM, respectively. Finally, the damage ratio of mtDNA and nDNA was assessed by quantitative PCR. RESULTS: MTT assay revealed that the HepG2 cells viabilities were significantly inhibited by olaquindox in dose- and time- dependent manners. Olaquindox induced increased levels of ROS and calcium in HepG2 cells, and reduced the level of △Ψm . Quantitative PCR assay showed that olaquindox led to a dose-dependent decrease in the amplification of both mtDNA and nDNA. These data also suggested that the olaquindox-induced damage to mtDNA was more extensive than its damage to nDNA. CONCLUSION: Olaquindox could cause oxidative damage of mitochondria in HepG2 cells.

Key words: olaquindox, mitochondria, reactive oxygen species, mitochondrial membrane potential, calcium

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