Copper Nanoparticles and Copper Sulphate Induced Cytotoxicity in Hepatocyte Primary Cultures of Epinephelus coioides

Copper nanoparticles (Cu-NPs) were widely used in various industrial and commercial applications. The aim of this study was to analyze the cytotoxicity of Cu-NPs on primary hepatocytes of E . coioides compared with copper sulphate (CuSO 4 ). Cultured cells were exposed to 0 or 2.4 mg Cu L -1 as CuSO 4 or Cu-NPs for 24-h. Results showed either form of Cu caused a dramatic loss in cell viability, more so in the CuSO 4 than Cu-NPs treatment. Compared to control, either CuSO 4 or Cu-NPs significantly increased reactive oxygen species(ROS) and malondialdehyde(MDA) concentration in hepatocytes by overwhelming total superoxide dismutase (T-SOD) activity, catalase(CAT) activity and glutathione(GSH) concentration. In addition, the antioxidative-related genes [ SOD (Cu/Zn) , SOD (Mn) , CAT , GPx4 ] were also down-regulated. The apoptosis and necrosis percentage was significantly higher after the CuSO 4 or Cu-NPs treatment than the control. The apoptosis was induced by the increased cytochrome c concentration in the cytosol and elevated caspase-3, caspase-8 and caspase-9 activities. Additionally, the apoptosis-related genes ( p53 , p38β and TNF-α ) and protein (p53 protein) were up-regulated after the CuSO 4 or Cu-NPs treatment, with CuSO 4 exposure having a greater effect than Cu-NPs. In conclusion, Cu-NPs had similar types of toxic effects as CuSO 4 on primary hepatocytes of E . coioides , but toxicity of CuSO 4 was more severe than that of Cu-NPs.