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Differences in propionate-induced inhibition of cholesterol and triacylglycerol synthesis between human and rat hepatocytes in primary culture

Published online by Cambridge University Press:  09 March 2007

Yuguang Lin ,
Roel J. Vonk ,
Maarten J. H. Slooff ,
Folkert Kuipers  and
Martin J. Smit
Yuguang Lin
Affiliation:
Groningen Institute for Drug Studies (GIDS), Department of PediatricsUniversity Hospital and State University of Groningen, The Netherlands
Roel J. Vonk
Affiliation:
Groningen Institute for Drug Studies (GIDS), Department of PediatricsUniversity Hospital and State University of Groningen, The Netherlands
Maarten J. H. Slooff
Affiliation:
Department of SurgeryUniversity Hospital and State University of Groningen, The Netherlands
Folkert Kuipers
Affiliation:
Groningen Institute for Drug Studies (GIDS), Department of PediatricsUniversity Hospital and State University of Groningen, The Netherlands
Martin J. Smit
Affiliation:
Groningen Institute for Drug Studies (GIDS), Department of PediatricsUniversity Hospital and State University of Groningen, The Netherlands
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Abstract

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Propionate is a short-chain fatty acid formed in the colon and supposedly involved in the cholesterol-lowering effect of soluble fibre. To explore the underlying mechanism(s) of this fibre action, we have used human hepatocytes in primary culture to study the effects of propionate on hepatic lipid synthesis. Initial experiments with mevalonate and mevinolin, a competitive inhibitor of hydroxymethylglutaryl (HMG)-CoA reductase (EC 1·1·1·88) were performed to evaluate basic regulatory mechanisms in these cells; results were compared with those obtained with rat hepatocytes. Incubation for 24 h with mevalonate caused a similar, concentration-dependent inhibition of [14C]acetate incorporation. into cholesterol in human and rat hepatocytes. Likewise, mevinolin (100 μmol/l) inhibited the formation of cholesterol from radiolabelled acetate by about 80% in cells from both species. Propionate inhibited cholesterol as well as triacylglycerol synthesis from [14C]acetate with a similar concentration-dependency in rat hepatocytes. Fifty percent inhibition was obtained at a propionate concentration of only 0·1 mmol/l· This propionate-induced inhibition was not affected by a 100-fold excess of unlabelled acetate. Human hepatocytes were much less susceptible in this respect: propionate concentrations of 10–20 mmol/l were required to obtain similar inhibitory effects in these cells, i.e. values greatly exceeding reported portal propionate concentrations in humans. The results suggest the existence of differences in the regulation of hepatic cholesterol (and triacylglycerol) synthesis between human and rat liver cells. These results do not support the hypothesis that the fibre-induced decrease in plasma cholesterol concentration in man is mediated by a direct effect of propionate on hepatic cholesterol synthesis.

Keywords

LiverCholesterolTriacylglycerolShort-chain fatty acids
Type
Effects of propionate in isolated hepatocytes
Information
British Journal of Nutrition , Volume 74 , Issue 2 , August 1995 , pp. 197 - 207
Copyright
Copyright © The Nutrition Society 1995

References

REFERENCES

Anderson, J. W., Story, L., Sieling, B., Chen, W., Petro, M. S. & Story, J. (1984) Hypocholesterolemic effects of oat-bran or bean intake for hypercholesterolemic men. American Journal of Clinical Nutrition 40, 1146–1155.Google Scholar
Berry, M. N. & Friend, D. S. (1969) High-yield preparation of isolated rat liver parenchymal cells. A biochemical and fine structural study. Journal of Cell Biology 43, 506–520.Google Scholar
Bligh, E. G. & Dyer, W. J. (1959) A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Biophysiology 37, 911–917.Google Scholar
Brass, E. P. & Ruff, L. J. (1992) Rat heptic coenzyme A is redistributed in response to mitochondrial acyl-coenzyme A accumulation. Journal of Nutrition 122, 2094–2100.Google Scholar
Chen, W. J. & Anderson, J. W. (1984) Propionate may mediate the hypocholesterolemic effects of certain soluble plant fibres in cholesterol-fed rats. Proceedings of the Society for Experimental Biology and Medicine 175, 215–218.Google Scholar
Cummings, J. H., Pomare, E. W., Branch, W. J., Naylor, C. P. E. & MacFarlane, G. T. (1987) Short chain fatty acids in human large intestine, portal, hepatic and venous blood. Gut 28, 1221–1227.Google Scholar
Dankert, J., Zijlstra, J. B. & Wolthers, B. G. (1981) Volatile fatty acids in human peripheral and portal blood: quantitative determination by vacuum distillation and gas chromatography. Clinica Chimica Acta 110, 301–307.Google Scholar
Davis, R. A., McNeal, M. M. & Moses, R. L. (1982) Intrahepatic assembly of very low density lipoprotein. Competition by cholesterol esters for the hydrophobic core. Journal of Biological Chemistry 257, 2634–2640.Google Scholar
De Water, R., Kamps, J. A. A., M., Van, Dijk, M. C. M., Hessels, E. M. A. J., Kuiper, J., Kruijt, J. K. & Van Berkel, T. J. C. (1992) Characterization of the low-density-lipoprotein-receptor-independent interaction of β-migrating very-low-density-lipoprotein with rat and human parenchymal liver cells in vitro. Biochemical Journal 282, 41–48.Google Scholar
Dietschy, J. (1986) Regulation of cholesterol metabolism in man and in other species. Klinische Wochenschrift 62, 338–345.Google Scholar
Drevon, C. A., Weinstein, D. B. & Steinberg, D. (1980) Regulation of cholesterol esterification and biosynthesis in monolayer cultures of normal adult rat hepatocytes. Journal of Biological Chemistry 225, 9128–9137.Google Scholar
Edge, S. B., Hoeg, J. M., Triche, T., Schneider, P. D. & Brewer, H. B. (1986) Cultured human hepatocytes. Evidence for metabolism of low density lipoproteins by a pathway independent of the classical low density lipoprotein receptor. Journal of Biological Chemistry 261, 3800–3806.Google Scholar
Forte, T. M., Nordhausen, R. W. & Princen, H. M. G. (1989) Structural properties of lipoproteins isolated from human primary hepatocyte cultures. Arteriosclerosis 9, 693 a.Google Scholar
Gamble, W., Vaughan, M., Kruth, M. S. & Avigan, J. (1978) Procedure for determination of free and total cholesterol in micro- or nanogram amounts suitable for studies with cultured cells. Journal of Lipid Research 19, 1068–1071.CrossRefGoogle Scholar
Goldstein, J. L. & Brown, M. S. (1980) Multivalent feedback regulation of HMG CoA reductase, a control mechanism coordinating isoprenoid synthesis and cell growth. Journal of Lipid Research 21, 505–517.Google Scholar
Gordon, M. J. & Crabtree, G. (1992) The effects of propionate and butyrate on acetate metabolism in rat hepatocytes. International Journal of Biochemistry 24, 1029–1032.Google Scholar
Havekes, L. M., Verboom, H., De Wit, E., Yap, S. H. & Princen, H. M. G. (1986) Regulation of low density lipoprotein receptor activity in primary cultures of human hepatocytes by serum lipoproteins. Hepatology 6, 1356–1360.Google Scholar
Hoeg, J. M., Edge, S. B., Demosky, S. J., Starzl, T. E., Triche, T., Gregg, R. E. & Brewer, H. B. (1986) Metabolism of low-density lipoproteins by cultured hepatocytes from normal and homozygous familial hypercholesterolemic subjects. Biochimica et Biophysica Acta 876, 646–657.Google Scholar
Illman, R. J., Topping, D. L., Mcintosh, G. H., Trimble, R. P., Storer, G. B., Taylor, M. N. & Cheng, B. Q. (1988) Hypocholesterolemic effects of dietary propionate; studies in whole animals and perfused rat liver. Annals of Nutrition and Metabolism 32, 97–107.Google Scholar
Kalayoglu, M., Sollinger, H. W., Stratta, R. J., D'Allessandro, A. M., Hoffman, R. M., Pirsch, J. D. & Belzer, F. O. (1988) Extended preservation of the liver for clinical transplantation. Lancet i, 617–619.Google Scholar
Kamps, J. A. A. M., Kruijt, J. K., Kuiper, J. & Van Berkel, T. J. C. (1991) Uptake and degradation of human low-density lipoprotein by human liver parenchymal and Kupffer cells in culture. Biochemical Journal 276, 135–140.Google Scholar
Kirby, R. W., Anderson, J. W. & Sieling, B. (1981) Oat-bran selectively lowers serum low-density lipoprotein concentrations of hypercholesterolemic men. American Journal of Clinical Nutrition 34, 824–829.CrossRefGoogle Scholar
Kritchevsky, D. (1986) Dietary fiber and atherosclerosis. In Dietary Fiber, Basic and Clinical Aspects, pp. 265–274, [Vahouny, G.V. and Kritchevsky, D., editors]. New York: Plenum Press.Google Scholar
Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. (1959) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193, 265–275.Google Scholar
Nakanishi, M., Goldstein, J. L. & Brown, M. S. (1988) Multivalent control of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Mevalonate-derived product inhibits translation of mRNA and accelerates degradation of enzyme. Journal of Biological Chemistry 263, 8929–8937.Google Scholar
Neese, R., Faix, D., Kletke, C, Wu, K., Wang, A. C, Shackleton, C. H. L. & Hellerstein, M. K. (1993) Measurements of endogenous synthesis of serum cholesterol in rats and humans using Mass Isotopomer Distribution Analysis (MIDA). American Journal of Physiology 264, E136–E147.Google Scholar
Nishina, P. M. & Freedland, R. A. (1990) Effects of propionate on lipid biosynthesis in isolated rat hepatocytes. Journal of Nutrition 120, 668–673.Google Scholar
Princen, H. M. G., Huijsmans, C. M. G., Kuipers, F., Vonk, R. J. & Kempen, H. J. M. (1986) Ketoconazole blocks bile acid synthesis in hepatocyte monolayer cultures and in vivo in rat by inhibiting cholesterol 7α-hydroxylase. Journal of Clinical Investigation 78, 1064–1071.Google Scholar
Salhanick, A. I., Schwartz, S. I. & Amatruda, J. M. (1991) Insulin inhibits apolipoprotein B secretion in isolated human hepatocytes. Metabolism 40, 275–279.Google Scholar
Sandker, G. W., Weert, B., Olinga, P., Wolters, H., Slooff, M. J. H., Meijer, D. K. F. & Groothuis, G. M. M. (1994) Characterization of transport in isolated human hepatocytes. A study with the bile acid taurocholic acid, the uncharged ouabain and the organic cations vecuronium and rocuronium. Biochemical Pharmacology 40, 2193–2200.Google Scholar
Schouten, D., Kleinherenbrink-Stins, M. F., Brouwer, A., Knook, D. L., Kamps, J. A. A. M., Kuiper, J. & Van Berkel, T. J. C. (1990) Characterization in vitro of interaction of human apolipoprotein E-free high density lipoprotein with human hepatocytes. Arteriosclerosis 10, 1127–1135.Google Scholar
Smit, M. J., Beekhuis, H., Duursma, A. M., Bouma, J. M. W. & Gruber, M. (1988) Catabolism of circulating enzymes. Plasma clearance, endocytosis, and breakdown of lactate dehydrogenase-1 in rabbits. Clinical Chemistry 34, 2475–2480.Google Scholar
Todesco, T., Rao, A. V., Bosello, O. & Jenkins, D. J. A. (1991) Propionate lowers blood glucose and alters lipid metabolism in healthy subjects. American Journal of Clinical Nutrition 54, 860–865.Google Scholar
Turley, S. D., Andersen, J. M. & Dietschy, J. M. (1981) Rates of sterol synthesis and uptake in the major organs of the rat in vivo. Journal of Lipid Research 22, 551–569.Google Scholar
Turley, S. D. & Dietschy, J. M. (1982) Cholesterol metabolism and excretion. In The Liver: Biology and Pathology, pp. 467–492 [Arias, I., Popper, H., Schachter, D. and Shafritz, D. A., editors]. New York: Raven Press.Google Scholar
Wright, R. S., Anderson, J. W. & Bridges, S. R. (1990) Propionate inhibits hepatocyte lipid synthesis. Proceedings of the Society for Experimental Biology and Medicine 195, 26–29.Google Scholar