癌变·畸变·突变 ›› 2010, Vol. 22 ›› Issue (5): 361-365.doi: 10.3969/j.issn.1004-616x.2010.05.008

• 论著 • 上一篇    下一篇

喹乙醇对HepG2细胞抗氧化系统的影响

张婷,陈倩,汤树生,靳溪,邹家杰,刘凤英,张燊,肖希龙   

  1. 中国农业大学动物医学院药理和毒理教研室, 北京 100193
  • 收稿日期:2010-03-25 修回日期:2010-05-13 出版日期:2010-09-30 发布日期:2010-09-30
  • 通讯作者: 肖希龙

Olaquindox weakened the antioxidant system in human hepatoma G2 cells

ZHANG Ting, CHEN Qian, TANG Shu-sheng, JIN Xi, ZOU Jia-jie, LIU Feng-ying, ZHANG Shen, XIAO Xi-long   

  1. Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agriculture University, Beijing 100193, China
  • Received:2010-03-25 Revised:2010-05-13 Online:2010-09-30 Published:2010-09-30
  • Contact: XIAO Xi-long

摘要: 目的: 观察喹乙醇对人肝癌细胞(HepG2)抗氧化系统的影响,探讨喹乙醇诱导HepG2细胞氧化性损伤的机制。 方法: 分别将0、200、400、800 μg/ml的喹乙醇作用于HepG2细胞24 h后,检测各组细胞内抗氧化酶/物的变化;另设800 μg/ml的喹乙醇作用于HepG2细胞3 h后,再用超氧化物歧化酶(SOD)(100 μg/ml)、过氧化氢酶(CAT)(100 μg/ml)及 SOD(100 μg/ml)+CAT(100 μg/ml)分别作用细胞3 h的各实验组,采用分子探针2’,7’_二氯荧光黄双乙酸盐(2,7_dichlorodihydrofluorescein diacetate, DCFDA)检测各组处理后细胞内活性氧含量的变化。 结果: 不同浓度的喹乙醇作用HepG2细胞24 h后,细胞总ATPase、Na+K+-ATPase 及 Ca2+Mg2+-ATPase 和细胞内SOD、谷胱甘肽(GSH)及谷胱甘肽过氧化物酶(GSH_PX)活性随喹乙醇浓度的升高而下降(P<0.05或P<0.01),且具有一定的剂量_效应关系,细胞内CAT活性较对照组下降(P<0.01),但无剂量_效应关系,而细胞内丙二醛(MDA)水平随喹乙醇浓度的升高而增加(P<0.05或P<0.01),且呈一定的剂量_效应关系。经喹乙醇(800 μg/ml)处理细胞后,再分别用CAT、SOD 及 SOD+CAT处理的各组,其ROS含量与单纯喹乙醇(800 μg/ml)组相比分别减少80%(P<0.01)、40%及95%(P<0.01)。 结论: 喹乙醇可使体外培养HepG2细胞的抗氧化系统受到一定的破坏,且产生的ROS形式以H2O2为主。

关键词: 喹乙醇, 超氧化物歧化酶, 过氧化氢酶, 谷胱甘肽, 三磷酸腺苷酶, 活性氧

Abstract: OBJECTIVE: To investigate the effect of olaquindox on the antioxidant system in human hepatoma G2(HepG2) cells. METHODS: HepG2 cells were treated with 0, 200, 400 and 800 μg/ml olaquindox in 24 h. The activities of superoxide dismutase(SOD) and catalase(CAT), the level of malondialdehyde(MDA), the activities of glutathione (GSH) and glutathione peroxidase (GSH_PX), Na+K+_ATPase and Ca2+Mg2+-ATPase were measured. SOD or CAT, and the combination of SOD+CAT were incubated for 3 h with olaquindox_treated HepG2 cells. The levels of reactive oxygen species (ROS) were evaluated by 2, 7_dichlorodihydrofluorescein diacetate(DCFDA). RESULTS: Olaquindox decreased the levels of T_ATPase, Na+K+-ATPase and Ca2+Mg2+-ATPase, SOD,GSH ,GSH_PX, in HepG2 cells(P<0.05 or P<0.01), with a dose_response relationship. After HepG2 cells were treated with olaquindox, the activity of CAT was decreased when compared with the control(P<0.01). Olaquindox led to a dose_dependent increase in the level of MDA(P<0.05 or P<0.01). While HepG2 cells were treated with olaquindox (800 μg/ml), CAT,SOD and SOD+CAT reduced ROS about 80% ,40%and 95%, respectively(P<0.01). CONCLUSION: Olaquindox disrupted the antioxidant system in HepG2 cells. Our results indicated that olaquindox_induced HepG2 cells predominantly generated H2O2, which could be scavenged by CAT.

Key words: olaquindox, superoxide dismutase, catalase, glutathione, ATPase, reactive oxygen species