硒拮抗氟致大鼠肝细胞氧化应激和DNA损伤作用  

Effects of selenium against fluoride-induced oxidative stress and DNA damage of rat hepatocytes

作　　者：段鹏[1] 陈晋[1] 杨波[2] 杨锐[2] 马丹[3] 李满[3] 李功波[3] 王建洲[1] 

机构地区：[1]湖北医药学院基础医学院,湖北十堰442000 [2]湖北医药学院第一临床医学院 [3]湖北医药学院第三临床医学院

出　　处：《中国公共卫生》2013年第11期1626-1629,共4页Chinese Journal of Public Health

基　　金：湖北医药学院研究生启动基金(2010QDJ16)

摘　　要：目的探讨硒对氟诱导的原代大鼠肝细胞氧化应激和DNA损伤的拮抗作用。方法采用改良原位2步非循环灌流法及多次过滤低速离心法分离原代大鼠肝细胞,实验设空白对照组、氟染毒组(氟化钠250、500、1 000、2 000μmol/L,染毒24 h)和硒干预组(亚硒酸钠10或100 nmol/L,干预4 h);测定细胞内活性氧(ROS)、丙二醛(MDA)、谷胱甘肽(GSH)含量及超氧化物歧化酶(SOD)活力,采用单细胞凝胶电泳技术和微核实验检测DNA损伤情况。结果与空白对照组比较,2 000μmol/L氟染毒组ROS(16.75±8.32)和MDA[(11.54±3.83)nmol/mg·prot]含量明显增加,SOD[(19.53±5.28)NU/mg·prot]和GSH[(21.73±15.32)μg/mg·prot]明显下降,彗星率[(46.25±10.60)%]、彗星尾长[(31.74±9.25)μm]、尾矩[(15.57±7.92)]和微核率[(42.80±24.61)‰]明显增加(均P<0.01);与等剂量氟染毒组比较,10 nmol/L硒干预组ROS(12.36±5.15)和MDA[(7.33±4.01)nmol/mg·prot]含量明显下降,SOD[(23.72±7.15)NU/mg·prot]与GSH[(26.36±14.24)μg/mg·prot]明显增加,彗星率[(39.27±15.0)%]、彗星尾长[(24.06±8.77)μm]、尾矩[(9.64±4.80)]和微核率[(31.28±12.65)‰]分别降低(P<0.05)。结论适量的硒可拮抗氟对原代大鼠肝细胞的氧化应激和DNA损伤作用。Objective To investigate the antagonism of selenium on oxidative stress and DNA damage induced by fluoride in primary hepatocytes of rats.Methods Primary hepatocytes of rats were isolated by using a modified two-step in-situ non-circulating perfusion method followed by multiple filtration and low-speed centrifugation.The blank control group,fluoride poisoning groups（250,500,1 000,2 000 μmol/L for 24 hours） and selenium intervening groups（10 or 100 nmol/L for 4 hours） were established.The contents of intracellular reactive oxygen species（ROS）,malondialdehyde（MDA）,glutathione（GSH）,and activities of superoxide dismutase（SOD） were measured.Meanwhile,the single cell gel electrophoresis（SCGE） and micronucleus test were adopted to detect DNA damage.Results Compared with the blank control group,at fluoride concentration of 2 000 μmol/L,the contents of ROS and MDA were increased to 16.78±8.32 and 11.54±3.83 nmol/ml·prot,whereas the activity of SOD and the content of GSH were decreased（19.53±5.28 NU/mg·port,21.73±15.32 μg/mg·prot）;the comet rate（46.25±10.60%）,comet tail length（31.74±9.25 μm）,tail moment（15.57±7.60）,and micronucleus rate（42.80±24.61‰） were also increased significantly（P〈0.01 for all）.In comparison with the fluoride poisoning group（2 000 μmol/L）,the contents of ROS and MDA in the 10 nmol/L selenium intervening group were decreased to 12.36±5.15 and 7.33±0.41 nmol/mg·prot,and the activity of SOD and the content of GSH were increased to 23.72±7.15 NU/mg·prot and 26.36±14.24 μg/mg·prot）.Moreover,the comet rate（39.27±15.09%）,comet tail length（24.06±8.77 μm）,tail moment（9.64±4.80）,and micronucleus rate（31.28±2.65‰） were signifiicantly increased（P〈0.05 for all）.Conclusion Selenium at proper dosage could antagonize oxidative stress and DNA damage induced by fluoride in primary hepatocytes of rats.

关 键 词：硒 氟 肝细胞 氧化应激 DNA损伤 

分 类 号：R995[医药卫生—毒理学]