FEED SCIENCE AND TECHNOLOGY

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

  • SONG Kai ,
  • LUO Yuan ,
  • ZHANG Chunxiao ,
  • WANG Ling ,
  • YOU Wenhuang ,
  • CHEN Xiaohui ,
  • YANG Da
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  • Key Laboratory of Healthy Mariculture for the East China Sea of Ministry of Agriculture, Key Laboratory for Feed Quality Testing and Safety Evaluation, Fish College, Jimei University, Xiamen 361021, China

Received date: 2016-05-11

  Online published: 2016-11-18

Abstract

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 (G6Pase), 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 G6Pase (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 of incubation, 100 and 1 000 nmol/L cortisol significantly increased the activity of hepatocyte PK (P<0.05). With the cortisol incubation time extending, the activity of hepatocyte G6Pase was significantly decreased (P<0.05), the activity of hepatocyte PK was significantly increased (P<0.05), however, there were no significant differences in the activities of hepatocyte MDH and ICD (P>0.05). The present study demonstrates that cortisol can enhance the gluconeogenesis, and has no regulating effect on synthesis of hepatic glycogen, but inhibits the catabolism of glucose of the primary cultured hepatocytes from Epinephelus coioides.

Cite this article

SONG Kai , LUO Yuan , ZHANG Chunxiao , WANG Ling , YOU Wenhuang , CHEN Xiaohui , YANG Da . Effects of Cortisol on Glycometabolism in Primary Cultured Hepatocytes form Epinephelus coioides[J]. Chinese Journal of Animal Nutrition, 2016 , 28(11) : 3520 -3527 . DOI: 10.3969/j.issn.1006-267x.2016.11.019

References

[1] IWAMA G K,VIJAYAN M M,FORSYTH R B,et al.Heat shock proteins and physiological stress in fish[J].American Zoologist,1999,39(6):901-909. 
[2] MOMMSEN T P,VIJAYAN M M,MOON T W.Cortisol in teleosts:dynamics,mechanisms of action,and metabolic regulation[J].Reviews in Fish Biology and Fisheries,1999,9(3):211-268. 
[3] GAAB J,ROHLEDER N,NATER U M,et al.Psychological determinants of the cortisol stress response:the role of anticipatory cognitive appraisal[J].Psychoneuroendocrinology,2005,30(6):599-610. 
[4] TORT L.Stress and immune modulation in fish[J].Developmental & Comparative Immunology,2011,35(12):1366-1375. 
[5] BARTON B A,IWAMA G K.Physiological changes in fish from stress in aquaculture with emphasis on the response and effects of corticosteroids[J].Annual Review of Fish Diseases,1991,1:3-26.
[6] WENDELAAR B S E.The stress response in fish[J].Physiological Reviews,1997,77(3):591-625.
[7] WILSON R P.Utilization of dietary carbohydrate by fish[J].Aquaculture,1994,124(1/2/3/4):67-80.
[8] LEUNG L Y,WOO N Y S.Effects of growth hormone,insulin-like growth factor I,triiodothyronine,thyroxine,and cortisol on gene expression of carbohydrate metabolic enzymes in sea bream hepatocytes[J].Comparative Biochemistry and Physiology Part A:Molecular & Integrative Physiology,2010,157(3):272-282. 
[9] PUVIANI A C,OTIOLENGHI C,GAVIOLI M E,et al.Action of glucagon and glucagon-like peptide on glycogen metabolism of trout isolated hepatocytes[J].Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,1990,96(2):387-391. 
[10] MOMMSEN T P,PLISETSKAYA E M.Insulin in fishes and agnathans:history,structure,and metabolic regulation[J].Reviews in Aquatic Sciences,1991,4(2/3):225-259.
[11] REINECKE M.Influences of the environment on the endocrine and paracrine fish growth hormone-insulin-like growth factor-Ⅰ system[J].Journal of Fish Biology,2010,76(6):1233-1254. 
[12] WOOD A W,DUAN C M,BERN H A.Insulin-like growth factor signaling in fish[J].International Review of Cytology,2005,243:215-285.
[13] BATTLE T,STACEY G.Cell culture models for hepatotoxicology[J].Cell Biology and Toxicology,2001,17(4/5):287-299.
[14] SEGNER H,BÖHM R,KLOAS W.Binding and bioactivity of insulin in primary cultures of carp (Cyprinus carpio) hepatocytes[J].Fish Physiology and Biochemistry,1993,11(1/2/3/4/5/6):411-420.
[15] QIN Q W,WU T H,JIA T L,et al.Development and characterization of a new tropical marine fish cell line from grouper,Epinephelus coioides susceptible to iridovirus and nodavirus[J].Journal of Virological Methods,2006,131(1):58-64. 
[16] KIKUCHI K,FURUTA T,HONDA H.Utilization of soybean meal as a protein source in the diet of juvenile japanese flounder,Paralichthys olivaceus[J].Aquaculture Science,1994,42(4):601-604.
[17] DE FRUTOS P G,BONAMUSA L,FERNANDEZ F,et al.Fructose 2,6-bisphosphate in liver of Sparus aurata:influence of nutritional state[J].Comparative Biochemistry and Physiology Part B:Comparative Biochemistry,1990,96(1):63-65. 
[18] VIJAYAN M M,MAULE A G,SCHRECK C B,et al.Hormonal control of hepatic glycogen metabolism in food-deprived,continuously swimming coho salmon (Oncorhynchus kisutch)[J].Canadian Journal of Fisheries and Aquatic Sciences,1993,50(8):1676-1682. 
[19] SUNNY F,LAKSHMY P S,OOMMEN O V.Rapid action of cortisol and testosterone on lipogenic enzymes in a fresh water fish Oreochromis mossambicus:short-term in vivo and in vitro study[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2002,131(3):297-304. 
[20] CHAN D K,WOO N Y S.Effect of hypophysectomy on the chemical composition and intermediary metabolism of the Japanese eel,Anguilla japonica[J].General and Comparative Endocrinology,1978,35(2):169-178. 
[21] SULLIVAN K M,SOMERO G N.Enzyme activities of fish skeletal muscle and brain as influenced by depth of occurrence and habits of feeding and locomotion[J].Marine Biology,1980,60(2/3):91-99.
[22] POLAKOF S,MÍGUEZ J M,SOENGAS J L.A hepatic protein modulates glucokinase activity in fish and avian liver:a comparative study[J].Journal of Comparative Physiology B:Biochemical,Systemic,and Environmental Physiology,2009,179(5):643-652. 
[23] BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry,1976,72(1/2):248-254.
[24] CHAN D K O,WOO N Y S.Effect of glucagon on the metabolism of the eal,Anguilla japonica[J].General and Comparative Endocrinology,1978,35(3):216-225. 
[25] FOSTER G D,MOON T W.Cortisol and liver metabolism of immature American eels,Anguilla rostrata (LeSueur)[J].Fish Physiology and Biochemistry,1986,1(2):113-124. 
[26] VIJAYAN M M,BALLANTYNE J S,LEATHERLAND J F.Cortisol-induced changes in some aspects of the intermediary metabolism of Salvelinus fontinalis[J].General and Comparative Endocrinology,1991,82(3):476-486. 
[27] VIJAYAN M M,LEATHERLAND J F.In vivo effects of the steroid analogue RU486 on some aspects of intermediary and thyroid metabolism of brook charr,Salvelinus fontinalis[J].Journal of Experimental Zoology,1992,263(3):265-271. 
[28] DAVE G,JOHANSSON-SJÖBECK M L,LARSSON Å,et al.Effects of insulin on the fatty acid composition of the total blood plasma lipids in the european EEL,Anguilla anguilla L[J].Comparative Biochemistry and Physiology Part A:Physiology,1979,62(3):649-653. 
[29] SHERIDAN M A.Effects of thyroxin,cortisol,growth hormone,and prolactin on lipid metabolism of coho salmon,Oncorhynchus kisutch,during smoltification[J].General and Comparative Endocrinology,1986,64(2):220-238. 
[30] VIJAYAN M M,FOSTER G D,MOON T W.Effects of cortisol on hepatic carbohydrate metabolism and responsiveness to hormones in the sea raven,Hemitripterus americanus[J].Fish Physiology and Biochemistry,1993,12(4):327-335. 
[31] MOMMSEN T P,MOON T W.Metabolic response of teleost hepatocytes to glucagon-like peptide and glucagon[J].Journal of Endocrinology,1990,126(1):109-118. 
[32] RADZIUK J,PYE S.Hepatic glucose uptake,gluconeogenesis and the regulation of glycogen synthesis[J].Diabetes/Metabolism Research and Reviews,2001,17(4):250-272. 
[33] VIJAYAN M M,MOON T W.The stress response and the plasma disappearance of corticosteroid and glucose in a marine teleost,the sea raven[J].Canadian Journal of Zoology,2011,72(3):379-386.
[34] VIJAYAN M M,REDDY P K,LEATHERLAND J F,et al.The effects of cortisol on hepatocyte metabolism in rainbow trout:a study using the steroid analogue RU486[J].General and Comparative Endocrinology,1994,96(1):75-84. 
[35] JONES C G,HOTHI S K,TITHERADGE M A.Effect of dexamethasone on gluconeogenesis,pyruvate kinase,pyruvate carboxylase and pyruvate dehydrogenase flux in isolated hepatocytes[J].The Biochemical Journal,1993,289(Pt 3):821-828.
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