中国水产科学2024，Vol.31Issue(5)：513-523,11.DOI:10.12264/JFSC2023-0359
高糖培养对花鲈肝细胞能量代谢与线粒体功能的影响
Effects of high-glucose load on energy metabolism and mitochondrial function of hepatocytes from spotted seabass(Lateolabrax maculatus)
摘要
Abstract
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.关键词
花鲈/离体肝细胞/高糖负荷/线粒体功能/糖代谢Key words
Lateolabrax maculatus/in vitro hepatocyte/high-glucose load/mitochondrial function/glycometabolism分类
农业科技引用本文复制引用
任惠娟,柯轶雄,鲁康乐,李学山,王玲,宋凯,张春晓..高糖培养对花鲈肝细胞能量代谢与线粒体功能的影响[J].中国水产科学,2024,31(5):513-523,11.基金项目
福建省杰出青年科学基金项目(2023J06035). （2023J06035）
