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. 1986 Apr 1;235(1):87–90. doi: 10.1042/bj2350087

Effect of prior nutritional status on the activity of lipogenic enzymes in primary monolayer cultures of rat hepatocytes.

D S Kelley, G J Nelson, J E Hunt
PMCID: PMC1146652  PMID: 2874793

Abstract

Effect of prior nutritional status of the animal on the activity of lipogenic enzymes and the fatty acid content of cultured hepatocytes was investigated. Hepatocytes were isolated from rats that were starved for 24 h ('starved') or continuously fed ('fed'), or starved for 48 h and then re-fed for 48 h ('re-fed') with a carbohydrate-rich fat-free diet, and maintained as monolayer cultures for 96 h in a serum-free glucose-rich medium (Waymouth's MB752/1) supplemented with insulin, dexamethasone and tri-iodothyronine. The fatty acid content and the activities of acetyl-CoA carboxylase, fatty acid synthase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were determined initially at 3 h after plating and then every 24 h. Initially the activities of all the four enzymes were highest in hepatocytes isolated from the re-fed rats and lowest in those from the starved rats. With time in culture, the activity of all these enzymes increased severalfold (2-5, depending on the enzyme under consideration) in hepatocytes isolated from fed and starved rats, whereas there was a severalfold (2-5) decrease in the activity of these enzymes in hepatocytes isolated from re-fed rats. The initial fatty acid content of the hepatocytes from re-fed rats was 2-3 times that in the other two groups of hepatocytes. The fatty acid content seemed to increase in all three groups of hepatocytes during the 96 h in culture, but these apparent increases were not statistically significant.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALLMANN D. W., HUBBARD D. D., GIBSON D. M. FATTY ACID SYNTHESIS DURING FAT-FREE REFEEDING OF STARVED RATS. J Lipid Res. 1965 Jan;6:63–74. [PubMed] [Google Scholar]
  2. Berke B. M., Kaplan M. L. Effects of high fat and high carbohydrate diets on development of hepatic and adipose lipogenesis in fa/fa and non-fa/fa rats. J Nutr. 1983 Apr;113(4):820–834. doi: 10.1093/jn/113.4.820. [DOI] [PubMed] [Google Scholar]
  3. Berry M. N., Friend D. S. High-yield preparation of isolated rat liver parenchymal cells: a biochemical and fine structural study. J Cell Biol. 1969 Dec;43(3):506–520. doi: 10.1083/jcb.43.3.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonney R. J., Becker J. E., Walker P. R., Potter V. R. Primary monolayer cultures of adult rat liver parenchymal cells suitable for study of the regulation of enzyme synthesis. In Vitro. 1974 May-Jun;9(6):399–413. doi: 10.1007/BF02615992. [DOI] [PubMed] [Google Scholar]
  5. Burton D. N., Collins J. M., Kennan A. L., Porter J. W. The effects of nutritional and hormonal factors on the fatty acid synthetase level of rat liver. J Biol Chem. 1969 Aug 25;244(16):4510–4516. [PubMed] [Google Scholar]
  6. Caro J. F., Amatruda J. M. The regulation of lipid synthesis in freshly isolated and primary cultures of hepatocytes from fasted rats: the primary role of insulin. Metabolism. 1982 Jan;31(1):14–18. [PubMed] [Google Scholar]
  7. Geelen M. J., Harris R. A., Beynen A. C., McCune S. A. Short-term hormonal control of hepatic lipogenesis. Diabetes. 1980 Dec;29(12):1006–1022. doi: 10.2337/diab.29.12.1006. [DOI] [PubMed] [Google Scholar]
  8. Gibbons G. F., Pullinger C. R., Björnsson O. G. Changes in the sensitivity of lipogenesis in rat hepatocytes to hormones and precursors over the diurnal cycle and during longer-term starvation of donor animals. J Lipid Res. 1984 Dec 1;25(12):1358–1367. [PubMed] [Google Scholar]
  9. Herzberg G. R. The influence of dietary fatty acid composition on lipogenesis. Adv Nutr Res. 1983;5:221–253. doi: 10.1007/978-1-4613-9937-7_10. [DOI] [PubMed] [Google Scholar]
  10. Inoue H., Lowenstein J. M. Acetyl coenzyme A carboxylase from rat liver. EC 6.4.1.2 acetyl-CoA: carbon dioxide ligase (ADP). Methods Enzymol. 1975;35:3–11. doi: 10.1016/0076-6879(75)35131-8. [DOI] [PubMed] [Google Scholar]
  11. Kelley D. S., Kletzien R. F. Ethanol modulation of the hormonal and nutritional regulation of glucose 6-phosphate dehydrogenase activity in primary cultures of rat hepatocytes. Biochem J. 1984 Jan 15;217(2):543–549. doi: 10.1042/bj2170543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kelley D. S., Potter V. R. Regulation of amino acid transport systems by amino acid depletion and supplementation in monolayer cultures of rat hepatocytes. J Biol Chem. 1978 Dec 25;253(24):9009–9017. [PubMed] [Google Scholar]
  13. Kelley D. S., Potter V. R. Repression, derepression, transinhibition, and trans-stimulation of amino acid transport in rat hepatocytes and four rat hepatoma cell lines in culture. J Biol Chem. 1979 Jul 25;254(14):6691–6697. [PubMed] [Google Scholar]
  14. Kim M., Elson C. E. Comparative studies of adaptive responses of fatty acid synthetase activities in rat liver and adipose tissue. J Nutr. 1981 Nov;111(11):1985–1995. doi: 10.1093/jn/111.11.1985. [DOI] [PubMed] [Google Scholar]
  15. Kurtz J. W., Wells W. W. Induction of glucose-6-phosphate dehydrogenase in primary cultures of adult rat hepatocytes. Requirement for insulin and dexamethasone. J Biol Chem. 1981 Nov 10;256(21):10870–10875. [PubMed] [Google Scholar]
  16. 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]
  17. Nakamura T., Yoshimoto K., Aoyama K., Ichihara A. Hormonal regulations of glucose-6-phosphate dehydrogenase and lipogenesis in primary cultures of rat hepatocytes. J Biochem. 1982 Feb;91(2):681–693. doi: 10.1093/oxfordjournals.jbchem.a133741. [DOI] [PubMed] [Google Scholar]
  18. Nepokroeff C. M., Lakshmanan M. R., Porter J. W. Fatty-acid synthase from rat liver. Methods Enzymol. 1975;35:37–44. doi: 10.1016/0076-6879(75)35136-7. [DOI] [PubMed] [Google Scholar]
  19. POTTER V. R., ONO T. Enzyme patterns in rat liver and Morris hepatoma 5123 during metabolic transitions. Cold Spring Harb Symp Quant Biol. 1961;26:355–362. doi: 10.1101/sqb.1961.026.01.043. [DOI] [PubMed] [Google Scholar]
  20. Sassoon H. F., Watson J., Johnson B. C. Diet-dependence of rat liver glucose 6-phosphate dehydrogenase levels. J Nutr. 1968 Jan;94(1):52–56. doi: 10.1093/jn/94.1.52. [DOI] [PubMed] [Google Scholar]
  21. TEPPERMAN H. M., TEPPERMAN J. The hexosemonophosphate shunt and adaptive hyperlipogenesis. Diabetes. 1958 Nov-Dec;7(6):478–485. doi: 10.2337/diab.7.6.478. [DOI] [PubMed] [Google Scholar]
  22. Taketa K., Pogell B. M. The effect of palmityl coenzyme A on glucose 6-phosphate dehydrogenase and other enzymes. J Biol Chem. 1966 Feb 10;241(3):720–726. [PubMed] [Google Scholar]
  23. Wakil S. J., Stoops J. K., Joshi V. C. Fatty acid synthesis and its regulation. Annu Rev Biochem. 1983;52:537–579. doi: 10.1146/annurev.bi.52.070183.002541. [DOI] [PubMed] [Google Scholar]
  24. Yugari Y., Matsuda T. [Glucose-6-phosphate dehydrogenase from rat liver. II. Effect of diet on enzyme activity in vivo, and inhibition by long chain fatty acids in vitro]. J Biochem. 1967 May;61(5):541–549. doi: 10.1093/oxfordjournals.jbchem.a128584. [DOI] [PubMed] [Google Scholar]

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