Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5757253 | Marine Pollution Bulletin | 2017 | 10 Pages |
Abstract
The present study aimed to use primary liver cell culture derived from the orange-spotted grouper, Epinephelus coioides, to assess the toxic effects of nonylphenol (NP) on the hepatocyte viability and the liver antioxidant system. E. coioides was selected due to its commercial importance. NP was used in this study because of its high potential of producing oxidative stress due to increased reactive oxygen species (ROS). A liver of E. coioides was digested with PBS containing 0.1% collagenase IV. The digested cells were moved to Leibovitz L-15 culture medium with 20% fetal bovine serum (FBS), 100 IU mLâ 1 penicillin, 100 μg mLâ 1 streptomycin. Aliquots of cell suspension were seeded as a monolayer into sterile 25 cm2 tissue culture flasks and incubated at 30 °C for 14 days. The medium, containing non-attached cells, was removed after 24 to 48 h and a new medium was added. The IC50 of 10â 4 mol Lâ 1 was determined for nonylphenol using MTT assay. Cells were then incubated with L-15 medium containing 10â 5, 2 Ã 10â 5, 3 Ã 10â 5 mol Lâ 1 of NP and samples were taken after 6, 12 and 24 h of incubation for analysis of LPO, SOD, CAT, GPx, LDH, AST, ALT, and ALP. Based on the results, the lowest concentration of NP was not markedly cytotoxic to primary hepatocytes and the cell sensitivity to NP increased dose-dependently. The activities of SOD, CAT and GPx decreased significantly, while activities of LPO, LDH, AST, ALT and ALP, increased significantly in a dose-related pattern in NP-treated cells. In conclusion, this study revealed that NP could induce the oxidative stress in cultivated hepatocytes of E. coioides during a short-term exposure. NP toxicity is mainly due to the induction of the reactive oxygen species (ROS), which lead to cell membrane disruption, damage of cellular metabolism, and interference with cellular macromolecules.
Keywords
FBSLiver cell cultureITSALTCATLPOGPXEpinephelus coioidesAPsDMSOASTAspartate aminotransferaseAlanine aminotransferaseALPAlkaline phosphataseAlkylphenolsOxidative stressDimethyl sulfoxideSODfetal bovine serumSuperoxide dismutaselactate dehydrogenaseLDHApesNonylphenolLipid peroxidationCatalaseglutathione peroxidase
Related Topics
Physical Sciences and Engineering
Earth and Planetary Sciences
Oceanography
Authors
Negin Derakhshesh, AbdolAli Movahedinia, Negin Salamat, Mahmoud Hashemitabar, Vahid Bayati,