摘要
目的探讨羟基酪醇对苏丹红Ⅰ号(SudanⅠ)所致的人肝癌细胞(HepG2)DNA氧化损伤的抑制作用以及作用机制。方法采用单细胞凝胶电泳(SCGE)检测细胞DNA氧化损伤;以21,71-二氢二氯荧光素二乙酸酯(DCFH-DA)为荧光探针检测细胞内活性氧(ROS)水平;用硫代巴比妥酸反应物(TBARS)法检测细胞内脂质过氧化水平;以免疫组化法检测细胞内8-羟基脱氧鸟苷(8-OHdG)的表达水平。结果100μmol/L苏丹红Ⅰ号引起HepG2细胞的DNA链断裂程度[尾长(46.49±2.06)μm,尾距(19.99±1.44)μm]、ROS水平〔荧光强度(9.03±1.05)〕、细胞内TBARS产物(吸光度0.41±0.15)及8-OHdG表达水平(相对染色密度90.4±1.40)比对照组(二甲基亚砜)均明显增加(P<0.01);不同浓度的羟基酪醇(0,25,50,100μmol/L)分别预处理HepG2细胞30 min后,再加入100μmol/L苏丹红Ⅰ号后,羟其酪醇预处理组各项指标较单独接触苏丹红Ⅰ号组明显降低(P<0.01或P<0.05),并且呈剂量依赖关系。结论羟基酪醇能够通过调控细胞氧化应激状态从而减轻苏丹红Ⅰ号所致的DNA氧化损伤。
Objective To explore the inhibitory effect of hydroxytyrosol （HT）on oxidative injury induced by Sudan Ⅰ in HepG2 cells. Methods The single cell gel electrophoresis assay （SCGE） was performed to study the DNA damage. The intracellular reactive oxygen species （ ROS ） formation was measured using 2,7-dichlorofluorescein diacetate （ DCFHDA） as a fluorescent probe. The levels of oxidative DNA damage and lipid peroxidation were estimated by immunocytochemistry analysis of 8-hydrocyde-oxyguanosine （8-OHdG） and by measuring the level of thtiobarbituric acid-reactive substances （TBARS）. Results Compared with the control group,HepG2 cells treated with 100 μM Sudan Ⅰ showed significantly different DNA strand breaks （ tail length of comet ：46.49 ± 2. 061μm, the tail moment： 19.99 ± 1. 44μm）, and the increases of ROS （9.03 ± 1. 05μm measured by fluorescent intensity） ,intracallular TBARS level（0.41 ±0. 15μm）measured by absorbency,and the increase of 8-OHdG expression（20. 5 ± 0. 09μm measured by rleative staining intensity）. Pretreatment with HT could inhibit the DNA damage and decrease the level of ROS , TBARS and 8-OHdG in a concentration-dependent manner. Conclusion HT could modulate the redox state and prevent the oxidative damage induced by Sudan Ⅰ in HepG2 cells.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2009年第9期1116-1117,共2页
Chinese Journal of Public Health
基金
大连市卫生局基金项目
