Effects of high-glucose load on energy metabolism and mitochondrial function of hepatocytes from spotted seabass(Lateolabrax maculatus)
This study aimed to investigate the metabolic characteristics of hepatocytes under a high-glucose load in spotted sea bass(Lateolabrax maculatus).The liver cell line of L.maculatus was cultured,and two experimental groups were established.The cells from the control group were cultured in a medium containing 5 mmol/L glucose,whereas a medium containing 40 mmol/L glucose was used for the high-glucose group.After 48 h of culture,the cells and supernatants were collected and analyzed.Results showed that the glucose and glycogen contents in the high-glucose group were significantly increased(P<0.05).The activities of hexokinase(HK),citrate synthase(CS),α-ketoglutarate dehydrogenase(α-KGDHC),isocitrate dehydrogenase(IDH),and succinate dehydrogenase(SDH)increased significantly(P<0.05).These results indicate that a high-glucose load accelerates glycolysis and the tricarboxylic acid cycle in hepatocytes.In addition,the activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the supernatant increased with high-glucose culture(P<0.05),indicating that high-glucose can cause hepatocyte damage.In the high-glucose group,the ATP content of hepatocytes significantly decreased(P<0.05).The content of reactive oxygen species(ROS)and MDA was significantly increased(P<0.05).Furthermore,the high-glucose culture upregulated the expressions of mitochondrial autophagy-related genes(pink,atg5,and mull)and down-regulated the expressions of mitochondrial biogenesis-related genes(nrf-1,pgc-1α,and pgc-1β)(P<0.05).Sequencing of the mitochondrial genome showed that mitochondrial D-loop genes were mutated under a high-glucose load treatment.In conclusion,in vitro hepatocytes of sea bass showed similar physiological responses to those in vivo when they responded to high-glucose loads.Thus,in vitro hepatocytes could be used as a research platform to study glucose metabolism in fish.High-glucose cultures can lead to increased metabolic enzyme activity in hepatocytes,damaging hepatocytes and mitochondria.This is related to the oxidative stress caused by ROS accumulation.The high-glucose load upregulated the expression of mitochondrial autophagy-related genes and downregulated the expression of mitochondrial generation-related genes,decreasing the number of mitochondria in the cell.Mutations in the mitochondrial D-loop gene occur under a high-glucose load,affecting the structural stability of the mitochondria.
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