Ammonium induces oxidative stress, endoplasmic reticulum stress, and apoptosis of hepatocytes in the liver cell line of grass carp (Ctenopharyngodon idella).

Abstract

Under unfavorable agricultural conditions, ammonia toxicity has become a major problem, resulting in a large number of deaths. Ammonia has been shown to be hepatotoxic. Research has also shown that ammonia can damage the livers of carp, but the mechanism is unclear. In this study, normal grass carp hepatocytes (L8824) were exposed to ammonia water to investigate the effect of ammonia on hepatocyte injury and apoptosis and its mechanism. The results showed that ammonia (50mM) reduced the viability of L8824 cells and increased glutamic pyruvic transaminase (ALT, up 144.95%, P < 0.01) and glutamic oxalacetic transaminase (AST, up 65.27%, P < 0.01). Furthermore, exposure to ammonia induced oxidative stress and endoplasmic reticulum (ER) stress in L8824 cells. Elevated levels of reactive oxygen species (ROS) and malondialdehyde (MDA) and decreased mitochondrial membrane potential indicated that L8824 cells suffered oxidative damage. Endoplasmic reticulum stress manifests as increased expression degrees of PERK, ATF4, and IRE-1α. These results confirmed the toxicity of ammonia to hepatocytes. In addition, the rate of apoptosis in L8824 cells was increased 69.66% after exposure to ammonia (50mM, P < 0.01). However, pretreatment of L8824 cells with ER stress inhibitor 2-APB reduced ammonia-induced calcium release (26.50%, P < 0.01) in endoplasmic reticulum. These results indicate that ammonia can exert toxic effects on L8824 cells through inducing endoplasmic reticulum stress and oxidative stress, resulting in apoptosis in L8824 cells.