Temporal Expression Pattern of Lipid Metabolism-related Genes and Reactive Oxygen Species (ROS) Level in Grass Carp (Ctenopharyngodon idellus) L8824 Cell Line during Hepatic Steatosis

Abstract

Objective: To investigate the mechanism of grass carp hepatic steatosis and the role of ROS in this process, grass carp liver cell line L8824 is used to establish the hepatic steatosis model by induction with oleic acid. Methods: The CCK8 kit and alanine transaminase (ALT) and aspartate transaminase (AST) were used to evaluate the cell variability and cell damage. Real time-PCR was used to detect the  key genes of lipid metabolism (PPAR, SREBP-1c, ACC, SCD-1, CPT-1 and MTTP). DCFH-DA probe was used to detect ROS. The results showed that treatment with oleic acid can continuously reduce the cell viability and exacerbate cell damage. The fatty acid de novo synthesis, β-oxidation a temporal expression pattern of lipid metabolism-related genes and Reactive Oxygen Species (ROS) level in grass carp (Ctenopharyngodon idellus) L8824 cell line during hepatic steatosisnd VLDL assemble were enhanced during the treatment from 0-24h, and then all the aspects were impaired by continuous induction. The ROS was maintained at a low level during 0-24h, then significantly increased at 36 h, then decreased to a second high level. Conclusion: The lipogenesis, β-oxidation, and triglyceride (TG) transportation are all heightened during nutritional steatosis until the hepatocytes fully loaded with fat, and the ROS levels perform in an opposite trend.