Using a liver cell culture from Epinephelus coioides as a model to evaluate the nonylphenol-induced oxidative stress. Issue 1 (15th September 2017)

• Record Type: Journal Article
• Authors: Derakhshesh, Negin; Movahedinia, AbdolAli; Salamat, Negin; Hashemitabar, Mahmoud; Bayati, Vahid
• Is Part Of: Marine pollution bulletin. Volume 122:Issue 1/2(2017)
• Page Start: 243
• Page End: 252
• Publication Date: 2017-09-15
• Subjects: L-15 culture medium Leibovitz L-15 culture medium -- NP nonylphenol -- DMSO dimethyl sulfoxide -- ITS insulin, transferrin, and selenium -- FBS fetal bovine serum -- SOD superoxide dismutase -- CAT catalase -- GPx glutathione peroxidase -- LPO lipid peroxidation -- ALT alanine aminotransferase -- AST aspartate aminotransferase -- ALP alkaline phosphatase -- LDH lactate dehydrogenase -- Aps alkylphenols -- APEs alkylphenols polythoxylates; Liver cell culture -- Epinephelus coioides -- Nonylphenol -- Oxidative stress; Marine pollution -- Periodicals; Marine Biology -- Periodicals; Water Pollution -- Periodicals; Mer -- Pollution -- Périodiques; Publications périodiques; Pollution des mers; Lutte antipollution; Electronic journals; 363.7394
• DOI: 10.1016/j.marpolbul.2017.06.049
• Languages: English
• ISSNs: 0025-326X

Abstract

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 mL − 1 penicillin, 100 μg mL − 1 streptomycin. Aliquots of cell suspension were seeded as a monolayer into sterile 25 cm 2 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 10 − 4 mol L − 1 was determined for nonylphenol using MTT assay. Cells were then incubated with L-15 medium containing 10 − 5, 2 × 10 − 5, 3 × 10 − 5 mol L − 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. Highlights: Epinephelus coioides primary hepatocyte culture was a suitable method to assess toxic effects of nonylphenol (NP). The IC50 of 10 − 4 M was determined for NP using MTT assay. The activities of SOD, CAT and GPx significantly decreased in hepatocytes treated with different concentrations of NP. The activities of LPO, LDH, AST, ALT and ALP significantly increased in a dose-related pattern in NP-treated cells. NP disrupts the antioxidant balance, which lead to oxidative stress in fish liver cell culture.