皮质醇对斜带石斑鱼原代培养肝细胞糖代谢的影响  

Effects of Cortisol on Glycometabolism in Primary Cultured Hepatocytes form Epinephelus coioides

作　　者：宋凯[1] 骆源[1] 张春晓[1] 王玲[1] 游文煌 陈晓卉 阳达 

机构地区：[1]农业部东海海水健康养殖重点实验室,厦门市集美大学饲料检测与安全评价重点实验室,集美大学水产学院,厦门361021

出　　处：《动物营养学报》2016年第11期3520-3527,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金青年科学基金项目(31302198);国家公益性行业(农业)专项(201303053);集美大学大学生创新性试验计划项目(201610390103)

摘　　要：本试验旨在研究皮质醇对斜带石斑鱼原代培养肝细胞糖代谢的影响。分离斜带石斑鱼肝细胞,选取2个孵育时间(24和36 h)和3个皮质醇浓度[0(对照)、100和1 000 nmol/L],测定培养上清液中葡萄糖含量(即肝细胞葡萄糖释放量),肝细胞糖原和丙酮酸含量以及肝细胞糖代谢关键酶[磷酸烯醇丙酮酸羧激酶(PEPCK)、葡萄糖-6-磷酸酶(G6Pase)、糖原合成酶(GSase)、苹果酸脱氢酶(MDH)、异柠檬酸脱氢酶(ICD)和丙酮酸激酶(PK)]的活性。结果表明:在孵育24和36 h时,与对照组相比,100和1 000 nmol/L皮质醇组肝细胞葡萄糖释放量显著提高(P<0.05),而肝细胞丙酮酸含量显著降低(P<0.05);皮质醇孵育时间对肝细胞葡萄糖释放量、丙酮酸含量均无显著影响(P>0.05)。在孵育24 h时,皮质醇浓度对肝细胞糖原含量无显著影响(P>0.05),而在孵育36 h时,肝细胞糖原含量则随着皮质醇浓度的升高而显著降低(P<0.05);随着皮质醇孵育时间的延长,肝细胞糖原含量显著下降(P<0.05)。在孵育24和36 h时,100和1 000 nmol/L皮质醇显著提高了肝细胞PEPCK和G6Pase的活性(P<0.05),显著降低了肝细胞MDH和ICD的活性(P<0.05),但对肝细胞GSase活性则无显著影响(P>0.05);在孵育24 h时,皮质醇浓度对肝细胞PK活性无显著影响(P>0.05),但在孵育36 h时,100和1 000 nmol/L皮质醇则显著提高了肝细胞PK活性(P<0.05)。随着孵育时间的延长,肝细胞G6Pase活性显著下降(P<0.05),PK活性显著升高(P<0.05),而MDH和ICD活性均无显著变化(P>0.05)。由此可见,皮质醇能够促进斜带石斑鱼原代培养肝细胞糖异生作用,抑制葡萄糖的分解代谢,而对糖原合成无调节作用。This experiment was conducted to study the effects of cortisol on glycometabolism in primary cultured hepatocytes form Epinephelus coioides. Freshly hepatocytes were isolated from Epinephelus coioides,and cultured at different cortisol concentrations [0（ control）,100 and 1 000 nmol / L] and different incubation time（ 24 and 36 h）. Medium glucose content（ hepatocyte glucose production）,hepatocyte glycogen and pyruvate contents,and the activities of glycometabolism related enzymes including phosphoenolpyruvate carboxykinase（ PECK）,glucose-6-phosphatase（ G6 Pase）,glycogen synthase（ GSase）,malate dehydrogenase（ MDH）,isocitrate dehydrogenase（ ICD） and pyruvate kinase（ PK） were assessed at 24 and 36 h of incubation,respectively. The results showed as follows： compared with the control group,the hepatocyte glucose production in 100 and 1 000 nmol / L cortisol groups was significantly increased（ P〈0. 05）,but the hepatocyte pyruvate content was significantly decreased（ P〈0. 05）,both at 24 and 36 h of incubation. The time of incubate to cortisol showed no significant effect on hepatocyte glucose production and pyruvate content（ P〉 0.05）. At 24 h of incubation,cortisol concentration showed no significant effect on hepatocyte glycogen content（ P〉0. 05）,however,at 36 h of incubation,hepatocyte glycogen content was significantly decreased with cortisol concentration increasing（ P〈0. 05）. With the cortisol incubation time extending,hepatocyte glycogen content was significantly decreased（ P〈0. 05）. At 24 and 36 h of incubation,100 and 1 000 nmol / L cortisol significantly increased the activities of hepatocyte PEPCK and G6 Pase（ P〈0. 05）,and significantly decreased the activities of hepatocyte MDH and ICD（ P〈0. 05）,however,there was no significant difference in the activity of hepatocyte GSase（ P〉0. 05）. At 24 h of incubation,cortisol concentration showed no significant effect on the activity of hepatocyte PK（ P〉0. 05）,but at 36 h o

关 键 词：斜带石斑鱼 原代培养 皮质醇 糖代谢 酶活性 

分 类 号：S963[农业科学—水产养殖]