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. 1996 Feb 15;314(Pt 1):103–108. doi: 10.1042/bj3140103

Chronic exogenous hyperinsulinaemia does not modify the acute inhibitory effect of insulin on the secretion of very-low-density lipoprotein triacylglycerol and apolipoprotein B in primary cultures of rat hepatocytes.

C S Bourgeois 1, D Wiggins 1, G F Gibbons 1
PMCID: PMC1217011  PMID: 8660269

Abstract

Male Wistar rats were fitted with subcutaneous osmotic mini-pumps that delivered insulin at a constant rate of 0.20 i.u./h for 7 days. This treatment raised the plasma insulin concentration from 31 +/- 4 to 201 +/- 64 micro-i.u./ml. Hepatocytes prepared from the hyperinsulinaemic animals secreted very-low-density lipoprotein (VLDL) triacylglycerol (TAG) at a higher rate (172 +/- 21 microgram per 24 h per mg cell protein) than did those from sham-operated controls (109 +/- 12 microgram per 24 h per mg) (P<0.05). However, chronic exogenous hyperinsulinaemia had no stimulatory effect on the secretion of VLDL apolipoprotein B (apoB) in derived hepatocytes compared with those from the sham-operated controls (2.32 +/- 0.38 compared with 3.09 +/- 0.40 microgram per 24 h per mg). Hepatocytes from the hyperinsulinaemic rats thus secreted larger VLDL particles as evidenced by the increased TAG:apoB ratio (78.4 +/- 13.1 compared with 38.4 +/- 7.6; P<0.05). In hepatocytes from the hyperinsulinaemic rats a larger proportion of the newly synthesized TAG was secreted as VLDL. Hepatocytes from the hyperinsulinaemic and the sham-operated control animals were equally sensitive to the inhibitory effect of insulin added in vitro on the secretion of VLDL TAG. Insulin added in vitro to the culture medium of hepatocytes from hyperinsulinaemic animals significantly decreased the TAG:apoB ratio of the secreted VLDL. This change did not occur in hepatocytes from sham-operated rats. These results suggest that, in vivo, chronic hyperinsulinaemia is not in itself sufficient to desensitize the liver to the acute inhibitory effect of insulin on the secretion of VLDL.

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Selected References

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  1. Abbott W. G., Swinburn B., Ruotolo G., Hara H., Patti L., Harper I., Grundy S. M., Howard B. V. Effect of a high-carbohydrate, low-saturated-fat diet on apolipoprotein B and triglyceride metabolism in Pima Indians. J Clin Invest. 1990 Aug;86(2):642–650. doi: 10.1172/JCI114756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Azain M. J., Fukuda N., Chao F. F., Yamamoto M., Ontko J. A. Contributions of fatty acid and sterol synthesis to triglyceride and cholesterol secretion by the perfused rat liver in genetic hyperlipemia and obesity. J Biol Chem. 1985 Jan 10;260(1):174–181. [PubMed] [Google Scholar]
  3. 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]
  4. Björnsson O. G., Duerden J. M., Bartlett S. M., Sparks J. D., Sparks C. E., Gibbons G. F. 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. Biochem J. 1992 Jan 15;281(Pt 2):381–386. doi: 10.1042/bj2810381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boden G., Chen X., Ruiz J., White J. V., Rossetti L. Mechanisms of fatty acid-induced inhibition of glucose uptake. J Clin Invest. 1994 Jun;93(6):2438–2446. doi: 10.1172/JCI117252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bonora E., Bonadonna R. C., Del Prato S., Gulli G., Solini A., Matsuda M., DeFronzo R. A. In vivo glucose metabolism in obese and type II diabetic subjects with or without hypertension. Diabetes. 1993 May;42(5):764–772. doi: 10.2337/diab.42.5.764. [DOI] [PubMed] [Google Scholar]
  7. Bourgeois C. S., Wiggins D., Hems R., Gibbons G. F. VLDL output by hepatocytes from obese Zucker rats is resistant to the inhibitory effect of insulin. Am J Physiol. 1995 Aug;269(2 Pt 1):E208–E215. doi: 10.1152/ajpendo.1995.269.2.E208. [DOI] [PubMed] [Google Scholar]
  8. Chang T. H., Polakis S. E. Differentiation of 3T3-L1 fibroblasts to adipocytes. Effect of insulin and indomethacin on the levels of insulin receptors. J Biol Chem. 1978 Jul 10;253(13):4693–4696. [PubMed] [Google Scholar]
  9. Cusin I., Rohner-Jeanrenaud F., Terrettaz J., Jeanrenaud B. Hyperinsulinemia and its impact on obesity and insulin resistance. Int J Obes Relat Metab Disord. 1992 Dec;16 (Suppl 4):S1–11. [PubMed] [Google Scholar]
  10. Cusin I., Terrettaz J., Rohner-Jeanrenaud F., Jeanrenaud B. Metabolic consequences of hyperinsulinaemia imposed on normal rats on glucose handling by white adipose tissue, muscles and liver. Biochem J. 1990 Apr 1;267(1):99–103. doi: 10.1042/bj2670099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davis R. A., Boogaerts J. R., Borchardt R. A., Malone-McNeal M., Archambault-Schexnayder J. Intrahepatic assembly of very low density lipoproteins. Varied synthetic response of individual apolipoproteins to fasting. J Biol Chem. 1985 Nov 15;260(26):14137–14144. [PubMed] [Google Scholar]
  12. 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]
  13. Duerden J. M., Gibbons G. F. Restoration in vitro of normal rates of very-low-density lipoprotein triacylglycerol and apoprotein B secretion in hepatocyte cultures from diabetic rats. Biochem J. 1993 Aug 15;294(Pt 1):167–171. doi: 10.1042/bj2940167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. Gibbons G. F., Bartlett S. M., Sparks C. E., Sparks J. D. Extracellular fatty acids are not utilized directly for the synthesis of very-low-density lipoprotein in primary cultures of rat hepatocytes. Biochem J. 1992 Nov 1;287(Pt 3):749–753. doi: 10.1042/bj2870749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gibbons G. F., Burnham F. J. Effect of nutritional state on the utilization of fatty acids for hepatitic triacylglycerol synthesis and secretion as very-low-density lipoprotein. Biochem J. 1991 Apr 1;275(Pt 1):87–92. doi: 10.1042/bj2750087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gibbons G. F. Hyperlipidaemia of diabetes. Clin Sci (Lond) 1986 Nov;71(5):477–486. doi: 10.1042/cs0710477. [DOI] [PubMed] [Google Scholar]
  20. Gibbons G. F. Insulin, diabetes and hepatic very-low-density lipoprotein metabolism. Biochem Soc Trans. 1989 Feb;17(1):49–51. doi: 10.1042/bst0170049. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Howard B. V., Abbott W. G., Egusa G., Taskinen M. R. Coordination of very low-density lipoprotein triglyceride and apolipoprotein B metabolism in humans: effects of obesity and non-insulin-dependent diabetes mellitus. Am Heart J. 1987 Feb;113(2 Pt 2):522–526. doi: 10.1016/0002-8703(87)90625-9. [DOI] [PubMed] [Google Scholar]
  23. Howard B. V. Lipoprotein metabolism in diabetes mellitus. J Lipid Res. 1987 Jun;28(6):613–628. [PubMed] [Google Scholar]
  24. Kissebah A. H., Schectman G. Hormones and lipoprotein metabolism. Baillieres Clin Endocrinol Metab. 1987 Aug;1(3):699–725. doi: 10.1016/s0950-351x(87)80029-0. [DOI] [PubMed] [Google Scholar]
  25. Kostner G. M., Karádi I. Lipoprotein alterations in diabetes mellitus. Diabetologia. 1988 Oct;31(10):717–722. doi: 10.1007/BF00274772. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Lewis G. F., Uffelman K. D., Szeto L. W., Steiner G. Effects of acute hyperinsulinemia on VLDL triglyceride and VLDL apoB production in normal weight and obese individuals. Diabetes. 1993 Jun;42(6):833–842. doi: 10.2337/diab.42.6.833. [DOI] [PubMed] [Google Scholar]
  28. Lewis G. F., Zinman B., Uffelman K. D., Szeto L., Weller B., Steiner G. VLDL production is decreased to a similar extent by acute portal vs. peripheral venous insulin. Am J Physiol. 1994 Oct;267(4 Pt 1):E566–E572. doi: 10.1152/ajpendo.1994.267.4.E566. [DOI] [PubMed] [Google Scholar]
  29. Livingston J. N., Purvis B. J., Lockwood D. H. Insulin induced changes in insulin binding and insulin-sensitivity of adipocytes. Metabolism. 1978 Dec;27(12 Suppl 2):2009–2014. doi: 10.1016/s0026-0495(78)80017-1. [DOI] [PubMed] [Google Scholar]
  30. Livingston J. N., Purvis B. J., Lockwood D. H. Insulin-dependent regulation of the insulin-sensitivity of adipocytes. Nature. 1978 Jun 1;273(5661):394–396. doi: 10.1038/273394a0. [DOI] [PubMed] [Google Scholar]
  31. Martin M. S., Pohl S. L. Insulin-induced insulin resistance of alpha-aminoisobutyric acid transport in cultured human skin fibroblasts. J Biol Chem. 1979 Oct 25;254(20):9976–9978. [PubMed] [Google Scholar]
  32. McGarry J. D. What if Minkowski had been ageusic? An alternative angle on diabetes. Science. 1992 Oct 30;258(5083):766–770. doi: 10.1126/science.1439783. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Reaven G. M. Banting lecture 1988. Role of insulin resistance in human disease. Diabetes. 1988 Dec;37(12):1595–1607. doi: 10.2337/diab.37.12.1595. [DOI] [PubMed] [Google Scholar]
  35. Sniderman A. D., Cianflone K. Metabolic disruptions in the adipocyte-hepatocyte fatty acid axis as causes of HyperapoB. Int J Obes Relat Metab Disord. 1995 May;19 (Suppl 1):S27–S33. [PubMed] [Google Scholar]
  36. Sparks J. D., Sparks C. E. Insulin regulation of triacylglycerol-rich lipoprotein synthesis and secretion. Biochim Biophys Acta. 1994 Nov 17;1215(1-2):9–32. doi: 10.1016/0005-2760(94)90088-4. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Sparks J. D., Sparks C. E. Obese Zucker (fa/fa) rats are resistant to insulin's inhibitory effect on hepatic apo B secretion. Biochem Biophys Res Commun. 1994 Nov 30;205(1):417–422. doi: 10.1006/bbrc.1994.2681. [DOI] [PubMed] [Google Scholar]
  39. Thorngate F. E., Raghow R., Wilcox H. G., Werner C. S., Heimberg M., Elam M. B. Insulin promotes the biosynthesis and secretion of apolipoprotein B-48 by altering apolipoprotein B mRNA editing. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5392–5396. doi: 10.1073/pnas.91.12.5392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Trimble E. R., Weir G. C., Gjinovci A., Assimacopoulos-Jeannet F., Benzi R., Renold A. E. Increased insulin responsiveness in vivo and in vitro consequent to induced hyperinsulinemia in the rat. Diabetes. 1984 May;33(5):444–449. doi: 10.2337/diab.33.5.444. [DOI] [PubMed] [Google Scholar]
  41. Vogelberg K. H., Gries F. A., Moschinski D. Hepatic production of VLDL-triglycerides. Dependence of portal substrate and insulin concentration. Horm Metab Res. 1980 Dec;12(12):688–694. doi: 10.1055/s-2007-999233. [DOI] [PubMed] [Google Scholar]
  42. Wardzala L. J., Hirshman M., Pofcher E., Horton E. D., Mead P. M., Cushman S. W., Horton E. S. Regulation of glucose utilization in adipose cells and muscle after long-term experimental hyperinsulinemia in rats. J Clin Invest. 1985 Aug;76(2):460–469. doi: 10.1172/JCI111994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wiggins D., Hems R., Gibbons G. F. Decreased sensitivity to the inhibitory effect of insulin on the secretion of very-low-density lipoprotein in cultured hepatocytes from fructose-fed rats. Metabolism. 1995 Jul;44(7):841–847. doi: 10.1016/0026-0495(95)90235-x. [DOI] [PubMed] [Google Scholar]
  44. Windmueller H. G., Spaeth A. E. Regulated biosynthesis and divergent metabolism of three forms of hepatic apolipoprotein B in the rat. J Lipid Res. 1985 Jan;26(1):70–81. [PubMed] [Google Scholar]
  45. 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]
  46. Yamamoto M., Yamamoto I., Tanaka Y., Ontko J. A. Fatty acid metabolism and lipid secretion by perfused livers from rats fed laboratory stock and sucrose-rich diets. J Lipid Res. 1987 Oct;28(10):1156–1165. [PubMed] [Google Scholar]

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