动物营养学报2017，Vol.29Issue(4)：1227-1232,6.DOI:10.3969/j.issn.1006-267x.2017.04.018
过氧化氢诱导斜带石斑鱼原代肝细胞氧化损伤模型的构建
Establishment of Oxidative Damage Model of Primary Hepatocytes of Grouper ( Epinephelus coioides) Induced by Hydrogen Peroxide
摘要
Abstract
This experiment was aimed to establish the stable oxidative damage model of primary hepatocytes of grouper using primary cultured hepatocytes of grouper ( Epinephelus coioides) as study object, hydrogen per-oxide (H2O2) as stress source and the changes of hepatocytes survival rate and antioxidant indexes as judgment indexes. The concentrations of 0 (control), 100, 200, 400, 600, 800 and 1000 μmol/L H2O2 were added in the cultured fluid of primary hepatocytes, and were cultured by 2, 4, 6, 8, 12 and 24 h, respectively. There were 42 groups and each group had 10 replicates. After culturing, the survival rate of hepatocytes was detected. Based on the appropriate action time was obtained, each concentration of H2O2(each concentration of H2O2 had 6 replicates) was used to culture hepatocytes for appropriate time. After culturing, the hepatocytes and cultured fluid were collected to determined antioxidant indexes, in order to select the appropriate action concentration of H2O2 ensuring oxidative damage to hepatocytes. The results showed that the survival rate of hepatocytes induced by 800μmol/L H2O2 at 8 h was reduced to 61.98%. The experiment also revealed that the 800 and 1000 μmol/L group was significantly decreased the activities of superoxide dismutase, glutathione peroxidase (except 600 μmol/L group) and catalase (P<0.05), and significantly increased the contents of malondialdehyde and lipid peroxidation compared with other groups ( P<0. 05 ) . However, the 800 and 1000 μmol/L groups had no significant differences (P>0.05). The results manifest that 800 μmol/L H2O2 incubated for 8 h can be used as a suitable method to establish oxidative damage model of primary hepatocytes of grouper.关键词
斜带石斑鱼/原代肝细胞/过氧化氢/氧化损伤模型Key words
grouper ( Epinephelus coioides )/primary hepatocytes/hydrogen peroxide/oxidative damage model分类
生物科学引用本文复制引用
张润蔚,王玲,张春晓,宋凯..过氧化氢诱导斜带石斑鱼原代肝细胞氧化损伤模型的构建[J].动物营养学报,2017,29(4):1227-1232,6.基金项目
福建省科技厅(重点)项目"石斑鱼保肝型饲料添加剂的研发与应用"(2016N0023) （重点）
国家自然科学基金青年基金项目"应用蛋白质组学技术研究皮质醇对斜带石斑鱼肝细胞代谢的影响机制"(31302198) （31302198）
福建省科技重大专项专题"石斑鱼健康养殖技术及高效配合饲料的研发和推广应用"(2016NZ0001-3) （2016NZ0001-3）
