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)

作　　者：任惠娟 柯轶雄 鲁康乐[1] 李学山 王玲[1] 宋凯[1] 张春晓[1] REN Huijuan;KE Yixiong;LU Kangle;LI Xueshan;WANG Ling;SONG Kai;ZHANG Chunxiao(State Key Laboratory of Mariculture Breeding,Fisheries College of Jimei University,Xiamen 361021,China)

机构地区：[1]集美大学水产学院,海水养殖生物育种全国重点实验室,福建厦门361021

出　　处：《中国水产科学》2024年第5期513-523,共11页Journal of Fishery Sciences of China

基　　金：福建省杰出青年科学基金项目(2023J06035).

摘　　要：为探究花鲈(Lateolabrax maculatus)肝细胞在高糖负荷下的代谢特征,本研究在正常糖(5 mmol/L,对照组)与高糖(40 mmol/L,高糖组)条件下培养花鲈肝脏细胞系,在培养48 h后,收集细胞及细胞培养基上清液测定相关指标。结果表明,与对照组相比,高糖组细胞糖原含量显著升高(P<0.05),且细胞己糖激酶、柠檬酸合酶、α-酮戊二酸脱氢酶、异柠檬酸脱氢酶、琥珀酸脱氢酶、苹果酸脱氢酶的活性显著提高(P<0.05),表明高糖负荷增强了肝细胞糖酵解和三羧酸循环。此外,高糖培养提高了上清液中谷丙转氨酶与谷草转氨酶的活性(P<0.05),表明高糖培养诱导肝细胞损伤。高糖负荷下,细胞活性氧(ROS)含量显著升高,ATP含量显著降低(P<0.05),并且在透射电镜下可观察到线粒体肿胀与嵴损伤,表明高糖诱导线粒体损伤。对线粒体自噬和合成相关基因表达分析发现,高糖培养显著上调了线粒体自噬相关基因pink、atg5、mul1的表达,并下调线粒体生成相关基因nrf-1、pgc-1α、pgc-1β的表达(P<0.05)。线粒体基因组测序表明,高糖培养肝细胞的线粒体D-Loop区基因发生了突变。综上所述,花鲈肝细胞通过增强糖酵解与三羧酸循环,促进线粒体自噬,抑制线粒体生成,诱导线粒体DNA突变以应对高糖带来的不良影响。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 t

关 键 词：花鲈 离体肝细胞 高糖负荷 线粒体功能 糖代谢 

分 类 号：S917[农业科学—水产科学]