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. 1992 Jan 15;281(Pt 2):381–386. doi: 10.1042/bj2810381

The role of pancreatic hormones in the regulation of lipid storage, oxidation and secretion in primary cultures of rat hepatocytes. Short- and long-term effects.

O G Björnsson 1, J M Duerden 1, S M Bartlett 1, J D Sparks 1, C E Sparks 1, G F Gibbons 1
PMCID: PMC1130695  PMID: 1310593

Abstract

Exposure of cultured rat hepatocytes to a high concentration of insulin (78 nM) for 24 h in the presence of extracellular oleate (0.75 mM) resulted in a decrease in the secretion of apoprotein B (apoB) and triacylglycerol associated with very-low-density lipoprotein (VLDL). However, continuous exposure of the cells to insulin for longer periods (72 h) stimulated the secretion of apoB and triacylglycerol. Treatment of hepatocytes with glucagon (0.1 microM) for 24 h also suppressed the secretion of VLDL apoB, cholesterol and triacylglycerol. The cells remained responsive to the inhibitory effect of glucagon for at least 3 days. In contrast with insulin, however, exposure of the cells to glucagon for a continuous period of 72 h did not lead to a reversal of the initial inhibition. Glucagon also stimulated ketogenesis, and in this regard the cells were responsive for at least 3 days in culture. These changes were accompanied by a transient increase in intracellular cyclic AMP (cAMP) concentration, which reached a peak 10 min after addition of glucagon. Between 12 h and 24 h after glucagon addition, cAMP levels had returned almost to normal, but the secretion of VLDL remained suppressed during this period.

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Selected References

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  1. Bartlett S. M., Gibbons G. F. Short- and longer-term regulation of very-low-density lipoprotein secretion by insulin, dexamethasone and lipogenic substrates in cultured hepatocytes. A biphasic effect of insulin. Biochem J. 1988 Jan 1;249(1):37–43. doi: 10.1042/bj2490037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry E. M., Ziv E., Bar-On H. Lipoprotein secretion by isolated perfused livers from streptozotocin-diabetic rats. Diabetologia. 1981 Oct;21(4):402–408. doi: 10.1007/BF00252689. [DOI] [PubMed] [Google Scholar]
  3. Brindle N. P., Ontko J. A. Alpha-adrenergic suppression of very-low-density-lipoprotein triacylglycerol secretion by isolated rat hepatocytes. Biochem J. 1988 Mar 1;250(2):363–368. doi: 10.1042/bj2500363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carling D., Zammit V. A., Hardie D. G. A common bicyclic protein kinase cascade inactivates the regulatory enzymes of fatty acid and cholesterol biosynthesis. FEBS Lett. 1987 Nov 2;223(2):217–222. doi: 10.1016/0014-5793(87)80292-2. [DOI] [PubMed] [Google Scholar]
  5. Dashti N., Williams D. L., Alaupovic P. Effects of oleate and insulin on the production rates and cellular mRNA concentrations of apolipoproteins in HepG2 cells. J Lipid Res. 1989 Sep;30(9):1365–1373. [PubMed] [Google Scholar]
  6. Davis R. A., Borchardt R. A. Phosphorylation-dephosphorylation of apolipoprotein B. Methods Enzymol. 1986;129:536–542. doi: 10.1016/0076-6879(86)29090-4. [DOI] [PubMed] [Google Scholar]
  7. Davis R. A., Clinton G. M., Borchardt R. A., Malone-McNeal M., Tan T., Lattier G. R. Intrahepatic assembly of very low density lipoproteins. Phosphorylation of small molecular weight apolipoprotein B. J Biol Chem. 1984 Mar 25;259(6):3383–3386. [PubMed] [Google Scholar]
  8. Duerden J. M., Bartlett S. M., Gibbons G. F. Long-term maintenance of high rates of very-low-density-lipoprotein secretion in hepatocyte cultures. A model for studying the direct effects of insulin and insulin deficiency in vitro. Biochem J. 1989 Nov 1;263(3):937–943. doi: 10.1042/bj2630937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duerden J. M., Bartlett S. M., Gibbons G. F. Regulation of very-low-density-lipoprotein lipid secretion in hepatocyte cultures derived from diabetic animals. Biochem J. 1989 Aug 15;262(1):313–319. doi: 10.1042/bj2620313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Duerden J. M., Gibbons G. F. Storage, mobilization and secretion of cytosolic triacylglycerol in hepatocyte cultures. The role of insulin. Biochem J. 1990 Dec 15;272(3):583–587. doi: 10.1042/bj2720583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Durrington P. N., Newton R. S., Weinstein D. B., Steinberg D. Effects of insulin and glucose on very low density lipoprotein triglyceride secretion by cultured rat hepatocytes. J Clin Invest. 1982 Jul;70(1):63–73. doi: 10.1172/JCI110604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Elovson J., Chatterton J. E., Bell G. T., Schumaker V. N., Reuben M. A., Puppione D. L., Reeve J. R., Jr, Young N. L. Plasma very low density lipoproteins contain a single molecule of apolipoprotein B. J Lipid Res. 1988 Nov;29(11):1461–1473. [PubMed] [Google Scholar]
  13. Fukuda N., Azain M. J., Ontko J. A. Altered hepatic metabolism of free fatty acids underlying hypersecretion of very low density lipoproteins in the genetically obese Zucker rats. J Biol Chem. 1982 Dec 10;257(23):14066–14072. [PubMed] [Google Scholar]
  14. Gibbons G. F. Assembly and secretion of hepatic very-low-density lipoprotein. Biochem J. 1990 May 15;268(1):1–13. doi: 10.1042/bj2680001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goldfarb S., Barber T. A., Pariza M. W., Pugh T. D. Lipid synthesis and ultrastructure of adult rat hepatocytes during their first twenty-four hours in culture. Exp Cell Res. 1978 Nov;117(1):39–46. doi: 10.1016/0014-4827(78)90425-1. [DOI] [PubMed] [Google Scholar]
  16. Hahn P. Development of lipid metabolism. Annu Rev Nutr. 1982;2:91–111. doi: 10.1146/annurev.nu.02.070182.000515. [DOI] [PubMed] [Google Scholar]
  17. Hahn P., Girard J., Assan R., Frohlich J., Kervran A. Control of blood cholesterol levels in suckling and weanling rats. J Nutr. 1977 Nov;107(11):2062–2066. doi: 10.1093/jn/107.11.2062. [DOI] [PubMed] [Google Scholar]
  18. Heimberg M., Weinstein I., Kohout M. The effects of glucagon, dibutyryl cyclic adenosine 3',5'-monophosphate, and concentration of free fatty acid on hepatic lipid metabolism. J Biol Chem. 1969 Oct 10;244(19):5131–5139. [PubMed] [Google Scholar]
  19. Heyworth C. M., Hanski E., Houslay M. D. Islet-activating protein blocks glucagon desensitization in intact hepatocytes. Biochem J. 1984 Aug 15;222(1):189–194. doi: 10.1042/bj2220189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoeg J. M., Meng M. S., Ronan R., Demosky S. J., Jr, Fairwell T., Brewer H. B., Jr Apolipoprotein B synthesized by Hep G2 cells undergoes fatty acid acylation. J Lipid Res. 1988 Sep;29(9):1215–1220. [PubMed] [Google Scholar]
  21. Huang G., Lee D. M., Singh S. Identification of the thiol ester linked lipids in apolipoprotein B. Biochemistry. 1988 Mar 8;27(5):1395–1400. doi: 10.1021/bi00405a001. [DOI] [PubMed] [Google Scholar]
  22. Jackson T. K., Salhanick A. I., Elovson J., Deichman M. L., Amatruda J. M. Insulin regulates apolipoprotein B turnover and phosphorylation in rat hepatocytes. J Clin Invest. 1990 Nov;86(5):1746–1751. doi: 10.1172/JCI114900. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  24. Mallinson C. N., Bloom S. R., Warin A. P., Salmon P. R., Cox B. A glucagonoma syndrome. Lancet. 1974 Jul 6;2(7871):1–5. doi: 10.1016/s0140-6736(74)91343-9. [DOI] [PubMed] [Google Scholar]
  25. Mangiapane E. H., Brindley D. N. Effects of dexamethasone and insulin on the synthesis of triacylglycerols and phosphatidylcholine and the secretion of very-low-density lipoproteins and lysophosphatidylcholine by monolayer cultures of rat hepatocytes. Biochem J. 1986 Jan 1;233(1):151–160. doi: 10.1042/bj2330151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mauger J. P., Claret M. Mobilization of intracellular calcium by glucagon and cyclic AMP analogues in isolated rat hepatocytes. FEBS Lett. 1986 Jan 20;195(1-2):106–110. doi: 10.1016/0014-5793(86)80140-5. [DOI] [PubMed] [Google Scholar]
  27. McGarry J. D., Meier J. M., Foster D. W. The effects of starvation and refeeding on carbohydrate and lipid metabolism in vivo and in the perfused rat liver. The relationship between fatty acid oxidation and esterification in the regulation of ketogenesis. J Biol Chem. 1973 Jan 10;248(1):270–278. [PubMed] [Google Scholar]
  28. Noda C., Shinjyo F., Tomomura A., Kato S., Nakamura T., Ichihara A. Mechanism of heterologous desensitization of the adenylate cyclase system by glucagon in primary cultures of adult rat hepatocytes. J Biol Chem. 1984 Jun 25;259(12):7747–7754. [PubMed] [Google Scholar]
  29. Patsch W., Gotto A. M., Jr, Patsch J. R. Effects of insulin on lipoprotein secretion in rat hepatocyte cultures. The role of the insulin receptor. J Biol Chem. 1986 Jul 25;261(21):9603–9606. [PubMed] [Google Scholar]
  30. Pullinger C. R., Gibbons G. F. Effects of hormones and pyruvate on the rates of secretion of very-low-density lipoprotein triacylglycerol and cholesterol by rat hepatocytes. Biochim Biophys Acta. 1985 Jan 9;833(1):44–51. doi: 10.1016/0005-2760(85)90251-6. [DOI] [PubMed] [Google Scholar]
  31. Pullinger C. R., North J. D., Teng B. B., Rifici V. A., Ronhild de Brito A. E., Scott J. The apolipoprotein B gene is constitutively expressed in HepG2 cells: regulation of secretion by oleic acid, albumin, and insulin, and measurement of the mRNA half-life. J Lipid Res. 1989 Jul;30(7):1065–1077. [PubMed] [Google Scholar]
  32. Reaven G. M., Greenfield M. S. Diabetic hypertriglyceridemia: evidence for three clinical syndromes. Diabetes. 1981;30(Suppl 2):66–75. doi: 10.2337/diab.30.2.s66. [DOI] [PubMed] [Google Scholar]
  33. Reaven G. M., Mondon C. E. Effect of in vivo plasma insulin levels on the relationship between perfusate free fatty acid concentration and triglyceride secretion by perfused rat livers. Horm Metab Res. 1984 May;16(5):230–232. doi: 10.1055/s-2007-1014753. [DOI] [PubMed] [Google Scholar]
  34. Shaheen O., Morgan D. W., Wilcox H. G., Keyes W. G., Heimberg M. Modulation by thyroid status of the actions of glucagon and dibutyryl adenosine 3',5'-monophosphate on metabolism of free fatty acids by the isolated perfused rat liver. Endocrinology. 1982 May;110(5):1740–1748. doi: 10.1210/endo-110-5-1740. [DOI] [PubMed] [Google Scholar]
  35. Sim A. T., Hardie D. G. The low activity of acetyl-CoA carboxylase in basal and glucagon-stimulated hepatocytes is due to phosphorylation by the AMP-activated protein kinase and not cyclic AMP-dependent protein kinase. FEBS Lett. 1988 Jun 20;233(2):294–298. doi: 10.1016/0014-5793(88)80445-9. [DOI] [PubMed] [Google Scholar]
  36. Sparks C. E., Sparks J. D., Bolognino M., Salhanick A., Strumph P. S., Amatruda J. M. Insulin effects on apolipoprotein B lipoprotein synthesis and secretion by primary cultures of rat hepatocytes. Metabolism. 1986 Dec;35(12):1128–1136. doi: 10.1016/0026-0495(86)90026-0. [DOI] [PubMed] [Google Scholar]
  37. Sparks J. D., Bolognino M., Trax P. A., Sparks C. E. The production and utility of monoclonal antibodies to rat apolipoprotein B lipoproteins. Atherosclerosis. 1986 Sep;61(3):205–211. doi: 10.1016/0021-9150(86)90139-5. [DOI] [PubMed] [Google Scholar]
  38. Sparks J. D., Sparks C. E., Bolognino M., Roncone A. M., Jackson T. K., Amatruda J. M. Effects of nonketotic streptozotocin diabetes on apolipoprotein B synthesis and secretion by primary cultures of rat hepatocytes. J Clin Invest. 1988 Jul;82(1):37–43. doi: 10.1172/JCI113597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sparks J. D., Sparks C. E. Insulin modulation of hepatic synthesis and secretion of apolipoprotein B by rat hepatocytes. J Biol Chem. 1990 May 25;265(15):8854–8862. [PubMed] [Google Scholar]
  40. Sparks J. D., Sparks C. E., Miller L. L. Insulin effects on apolipoprotein B production by normal, diabetic and treated-diabetic rat liver and cultured rat hepatocytes. Biochem J. 1989 Jul 1;261(1):83–88. doi: 10.1042/bj2610083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sparks J. D., Sparks C. E., Roncone A. M., Amatruda J. M. Secretion of high and low molecular weight phosphorylated apolipoprotein B by hepatocytes from control and diabetic rats. Phosphorylation of APO BH and APO BL. J Biol Chem. 1988 Apr 15;263(11):5001–5004. [PubMed] [Google Scholar]
  42. Steiner G., Haynes F. J., Yoshino G., Vranic M. Hyperinsulinemia and in vivo very-low-density lipoprotein-triglyceride kinetics. Am J Physiol. 1984 Feb;246(2 Pt 1):E187–E192. doi: 10.1152/ajpendo.1984.246.2.E187. [DOI] [PubMed] [Google Scholar]
  43. Unger R. H., Orci L. Physiology and pathophysiology of glucagon. Physiol Rev. 1976 Oct;56(4):778–826. doi: 10.1152/physrev.1976.56.4.778. [DOI] [PubMed] [Google Scholar]
  44. Van Harken D. R., Dixon C. W., Heimberg M. Hepatic lipid metabolism in experimental diabetes. V. The effect of concentration of oleate on metabolism of triglycerides and on ketogenesis. J Biol Chem. 1969 May 10;244(9):2278–2285. [PubMed] [Google Scholar]
  45. WILLIAMSON D. H., MELLANBY J., KREBS H. A. Enzymic determination of D(-)-beta-hydroxybutyric acid and acetoacetic acid in blood. Biochem J. 1962 Jan;82:90–96. doi: 10.1042/bj0820090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Weiland D., Mondon C. E., Reaven G. M. Evidence for multiple causality in the development of diabetic hypertriglyceridaemia. Diabetologia. 1980 Apr;18(4):335–340. doi: 10.1007/BF00251016. [DOI] [PubMed] [Google Scholar]
  47. Williamson D. H., Ilic V., Tordoff A. F., Ellington E. V. Interactions between vasopressin and glucagon on ketogenesis and oleate metabolism in isolated hepatocytes from fed rats. Biochem J. 1980 Feb 15;186(2):621–624. doi: 10.1042/bj1860621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Woodside W. F., Heimberg M. Effects of anti-insulin serum, insulin, and glucose on output of triglycerides and on ketogenesis by the perfused rat liver. J Biol Chem. 1976 Jan 10;251(1):13–23. [PubMed] [Google Scholar]

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