Molecular Nutrition

Establishment of Oxidative Damage Model of Primary Hepatocytes of Grouper (Epinephelus coioides) Induced by Hydrogen Peroxide

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  • Xiamen Key Laboratory for Feed Quality Testing and Safety Evaluation, Fisheries College, Jimei University, Xiamen 361021, China

Received date: 2016-10-25

  Online published: 2017-04-14

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 peroxide (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 1 000 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 800 μmol/L H2O2 at 8 h was reduced to 61.98%. The experiment also revealed that the 800 and 1 000 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 1 000 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.

Cite this article

ZHANG Runwei, WANG Ling, ZHANG Chunxiao, SONG Kai . Establishment of Oxidative Damage Model of Primary Hepatocytes of Grouper (Epinephelus coioides) Induced by Hydrogen Peroxide[J]. Chinese Journal of Animal Nutrition, 2017 , 29(4) : 1227 -1232 . DOI: 10.3969/j.issn.1006-267x.2017.04.018

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