LAUSR

Copper (Cu) is an essential trace element that is required for the catalysis of several cellular enzymes. Excessive Cu could induce hepatotoxicity in humans and multiple animals. The purpose of this study was to investigate the effects of autophagy machinery on Cu-induced hepatotoxicity. Chicken hepatocytes were cultured in medium in the absence and presence of Cu sulfate (CuSO4) (0, 10, 50, and 100 μM) for 0, 6, 12, and 24 h, and in the combination of CuSO4 and N-acetyl-l-cysteine (NAC) (1 mM), rapamycin (10 nM), and 3-methyladenine (3-MA) (5 mM) for 24 h. Results showed that Cu could markedly increase the number of autophagosomes and LC3 puncta, induce autophagy-related genes (Beclin1, ATG5, LC3Ⅰ, LC3Ⅱ, mTOR, and Dynein) mRNA expression and proteins (BECN1, LC3Ⅱ/LC3Ⅰ) expression. NAC could relieve Cu-induced the changes of above genes and proteins. Additionally, rapamycin attenuated Cu-induced the increased lactic dehydrogenase (LDH), aspartate amino transferase (AST), and alanine aminotransferase (ALT) activities, and SOD-1 mRNA expression as well as the decreased cell viability, reactive oxygen species (ROS), hydrogen peroxide, total superoxide dismutase (T-SOD), malonaldehyde (MDA), catalase (CAT), HO-1 mRNA expression, adenosine triphosphate (ATP) levels, mitochondrial mass, and mitochondria membrane potential (MMP). But 3-MA had the opposite effects on above factors. Collectively, these findings provide strong evidence that Cu could induce autophagy by generating excessive ROS in hepatocytes, and autophagy might attenuate Cu-induced mitochondrial dysfunction by regulating oxidative stress.