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. 1988 Dec 1;256(2):641–647. doi: 10.1042/bj2560641

Selective channelling of arachidonic and linoleic acids into glycerolipids of rat hepatocytes in primary culture.

G Thomas 1, C Loriette 1, D Pepin 1, J Chambaz 1, G Bereziat 1
PMCID: PMC1135457  PMID: 3223937

Abstract

Rat hepatocytes in primary culture were incubated with a mixture of linoleic and arachidonic acid at various total fatty acid/serum albumin molar ratios. Mixed fatty acids were taken up at the same rate and distributed with the same pattern as fatty acids added separately. The rates of total uptake, incorporation into hepatocyte and secreted triacylglycerols and beta-oxidation were linearly related to the fatty acid/albumin ratios, whereas the rate of incorporation into phospholipids was saturable. Neither the uptake rate nor the distribution of both fatty acids considered together varied with the arachidonic acid/linoleic acid molar ratio. Changes in this ratio and in the uptake rate led to significant variations in the respective fate of the fatty acids. The preferential channelling of arachidonic acid versus linoleic acid into beta-oxidation and phosphatidylinositol was greatest at a low uptake rate and then decreased as the uptake rose. Conversely, the preferential channelling of arachidonic acid versus linoleic acid into phosphatidylcholine, but not phosphatidylethanolamine, increased with the uptake rate. Moreover, both arachidonic acid and linoleic acid were preferentially incorporated into the 1-palmitoyl molecular species of phosphatidylcholine and phosphatidylethanolamine at a low uptake rate, and of phosphatidylcholine at a high uptake rate. This could be related to the synthesis of biliary phosphatidylcholine, of which 1-palmitoyl-2-linoleoyl and 1-palmitoyl-2-arachidonoyl are the main molecular species. Linoleic and arachidonic acid were selectively distributed into distinct metabolic pools of triacylglycerol, the intrahepatocyte pool which preferentially incorporated linoleic acid at a low uptake rate and the secreted pool in which the relative enrichment of arachidonic acid increased with the uptake rate. This strengthens the central role of hepatic secretion in the supply of arachidonic acid to peripheral tissues.

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Selected References

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

  1. Abumrad N. A., Stearns S. B., Tepperman H. M., Tepperman J. Studies on serum lipids, insulin, and glucagon and on muscle triglyceride in rats adapted to high-fat and high-carbohydrate diets. J Lipid Res. 1978 May;19(4):423–432. [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Balint J. A., Beeler D. A., Treble D. H., Spitzer H. L. Studies in the biosynthesis of hepatic and biliary lecithins. J Lipid Res. 1967 Sep;8(5):486–493. [PubMed] [Google Scholar]
  4. Beynen A. C., Vaartjes W. J., Geelen M. J. Acute effects of insulin on fatty acid metabolism in isolated rat hepatocytes. Horm Metab Res. 1980 Sep;12(9):425–430. doi: 10.1055/s-2007-999166. [DOI] [PubMed] [Google Scholar]
  5. Blank M. L., Robinson M., Fitzgerald V., Snyder F. Novel quantitative method for determination of molecular species of phospholipids and diglycerides. J Chromatogr. 1984 Aug 31;298(3):473–482. doi: 10.1016/s0021-9673(01)92744-x. [DOI] [PubMed] [Google Scholar]
  6. Chambaz J., Guillouzo A., Cardot P., Pepin D., Bereziat G. Essential fatty acid uptake and esterification in primary culture of rat hepatocytes. Biochim Biophys Acta. 1986 Oct 3;878(3):310–319. doi: 10.1016/0005-2760(86)90238-9. [DOI] [PubMed] [Google Scholar]
  7. Colard O., Breton M., Bereziat G. Hydrolysis of endogenous phospholipids by rat platelet phospholipase A2: ether or acyl bond and polar head group selectivity. Biochim Biophys Acta. 1987 Sep 25;921(2):333–340. doi: 10.1016/0005-2760(87)90034-8. [DOI] [PubMed] [Google Scholar]
  8. Groener J. E., van Golde L. M. Utilization of exogenously added and endogenously synthesized fatty acids for glycerolipids synthesis in isolated rat hepatocytes. Biochim Biophys Acta. 1978 Apr 28;529(1):88–95. doi: 10.1016/0005-2760(78)90106-6. [DOI] [PubMed] [Google Scholar]
  9. Guguen C., Guillouzo A., Boisnard M., Le Cam A., Bourel M. Etude ultrastructurale de monocouches d'hépatocytes de rat adulte cultivés en présence d'hémisuccinate d'hydrocortisone. Biol Gastroenterol (Paris) 1975 Jul-Aug;8(3):223–231. [PubMed] [Google Scholar]
  10. Hammarström S. Leukotrienes. Annu Rev Biochem. 1983;52:355–377. doi: 10.1146/annurev.bi.52.070183.002035. [DOI] [PubMed] [Google Scholar]
  11. Hasegawa-Sasaki H., Ohno K. Extraction and partial purification of acyl-CoA:1-acyl-sn-glycero-3-phosphocholine acyltransferase from rat liver microsomes. Biochim Biophys Acta. 1980 Feb 22;617(2):205–217. [PubMed] [Google Scholar]
  12. Holub B. J., MacNaughton J. A., Piekarski J. Synthesis of 1-palmitoyl and 1-stearoyl phosphatidylcholines from mixtures of acyl acceptors via acyl-CoA:1-acyl-sn-glycero-3-phosphorylcholine acyltransferase in liver microsomes. Biochim Biophys Acta. 1979 Mar 29;572(3):413–422. doi: 10.1016/0005-2760(79)90148-6. [DOI] [PubMed] [Google Scholar]
  13. Holub B. J. Specific formation of arachidonoyl phosphatidylinositol from 1-acyl-sn-glycero-3-phosphorylinositol in rat liver. Lipids. 1976 Jan;11(1):1–5. doi: 10.1007/BF02532576. [DOI] [PubMed] [Google Scholar]
  14. Holub B. J. The biosynthesis of phosphatidylserines by acylation of 1-acyl-sn-glycero-3-phosphoserine in rat liver. Biochim Biophys Acta. 1980 May 28;618(2):255–262. doi: 10.1016/0005-2760(80)90031-4. [DOI] [PubMed] [Google Scholar]
  15. Johnson S. B., Gordon E., McClain C., Low G., Holman R. T. Abnormal polyunsaturated fatty acid patterns of serum lipids in alcoholism and cirrhosis: arachidonic acid deficiency in cirrhosis. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1815–1818. doi: 10.1073/pnas.82.6.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kohout M., Kohoutova B., Heimberg M. The regulation of hepatic triglyceride metabolism by free fatty acids. J Biol Chem. 1971 Aug 25;246(16):5067–5074. [PubMed] [Google Scholar]
  17. 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]
  18. Lamb R. G., Wood C. K., Landa B. M., Guzelian P. S., Fallon H. J. Studies of the formation and relase of glycerolipids by primary monolayer cultures of adult rat hepatocytes. Biochim Biophys Acta. 1977 Nov 24;489(2):318–329. doi: 10.1016/0005-2760(77)90151-5. [DOI] [PubMed] [Google Scholar]
  19. Lands W. E., Inoue M., Sugiura Y., Okuyama H. Selective incorporation of polyunsaturated fatty acids into phosphatidylcholine by rat liver microsomes. J Biol Chem. 1982 Dec 25;257(24):14968–14972. [PubMed] [Google Scholar]
  20. Laposata M., Reich E. L., Majerus P. W. Arachidonoyl-CoA synthetase. Separation from nonspecific acyl-CoA synthetase and distribution in various cells and tissues. J Biol Chem. 1985 Sep 15;260(20):11016–11020. [PubMed] [Google Scholar]
  21. Lefkowith J. B., Flippo V., Sprecher H., Needleman P. Paradoxical conservation of cardiac and renal arachidonate content in essential fatty acid deficiency. J Biol Chem. 1985 Dec 15;260(29):15736–15744. [PubMed] [Google Scholar]
  22. Miki Y., Hosaka K., Yamashita S., Handa H., Numa S. Acyl-acceptor specificities of 1-acylglycerolphosphate acyltransferase and 1-acylglycerophosphorylcholine acyltransferase resolved from rat liver microsomes. Eur J Biochem. 1977 Dec;81(3):433–441. doi: 10.1111/j.1432-1033.1977.tb11968.x. [DOI] [PubMed] [Google Scholar]
  23. Neumüller W., Fleer E. A., Unger C., Eibl H. Enzymatic acylation of ether and ester lysophospholipids in rat liver microsomes. Lipids. 1987 Nov;22(11):808–812. doi: 10.1007/BF02535535. [DOI] [PubMed] [Google Scholar]
  24. Okuyama H., Yamada K., Ikezawa H. Accepton concentration effect in the selectivity of acyl coenzyme A: U aclglycerylphosphorylcholine acyltransferase system in rat liver. J Biol Chem. 1975 Mar 10;250(5):1710–1713. [PubMed] [Google Scholar]
  25. Ontko J. A. Metabolism of free fatty acids in isolated liver cells. Factors affecting the partition between esterification and oxidation. J Biol Chem. 1972 Mar 25;247(6):1788–1800. [PubMed] [Google Scholar]
  26. Owen J. S., Bruckdorfer K. R., Day R. C., McIntyre N. Decreased erythrocyte membrane fluidity and altered lipid composition in human liver disease. J Lipid Res. 1982 Jan;23(1):124–132. [PubMed] [Google Scholar]
  27. Parkes J. G., Chan P., Goldberg D. M. Secretion of triglyceride lipase from rat hepatocytes in culture: modulation by insulin and phenobarbital. Biochem Cell Biol. 1986 Nov;64(11):1147–1152. doi: 10.1139/o86-151. [DOI] [PubMed] [Google Scholar]
  28. Picard J., Veissiere D., Voyer F., Bereziat G. Composition en acids gras des phospholipdes dans les lipoproteines seriques anormales de la cholostase. Clin Chim Acta. 1972 Jan;36(1):247–250. doi: 10.1016/0009-8981(72)90184-2. [DOI] [PubMed] [Google Scholar]
  29. Saggerson E. D., Carpenter C. A. Effects of fasting, adrenalectomy and streptozotocin-diabetes on sensitivity of hepatic carnitine acyltransferase to malonyl CoA. FEBS Lett. 1981 Jul 6;129(2):225–228. doi: 10.1016/0014-5793(81)80170-6. [DOI] [PubMed] [Google Scholar]
  30. Skipski V. P., Smolowe A. F., Sullivan R. C., Barclay M. Separation of lipid classes by thin-layer chromatography. Biochim Biophys Acta. 1965 Oct 4;106(2):386–396. doi: 10.1016/0005-2760(65)90047-0. [DOI] [PubMed] [Google Scholar]
  31. Strum-Odin R., Adkins-Finke B., Blake W. L., Phinney S. D., Clarke S. D. Modification of fatty acid composition of membrane phospholipid in hepatocyte monolayer with n-3, n-6 and n-9 fatty acids and its relationship to triacylglycerol production. Biochim Biophys Acta. 1987 Sep 25;921(2):378–391. doi: 10.1016/0005-2760(87)90040-3. [DOI] [PubMed] [Google Scholar]
  32. Sundler R., Akesson B., Nilsson A. Effect of different fatty acids on glycerolipid synthesis in isolated rat hepatocytes. J Biol Chem. 1974 Aug 25;249(16):5102–5107. [PubMed] [Google Scholar]
  33. Weisiger R., Gollan J., Ockner R. Receptor for albumin on the liver cell surface may mediate uptake of fatty acids and other albumin-bound substances. Science. 1981 Mar 6;211(4486):1048–1051. doi: 10.1126/science.6258226. [DOI] [PubMed] [Google Scholar]

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