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In Vitro| Volume 24, ISSUE 4, P409-417, April 1983

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beta-sitosterol: esterification by intestinal acylcoenzyme A: cholesterol acyltransferase (ACAT) and its effect on cholesterol esterification

Open AccessPublished:April 01, 1983DOI:https://doi.org/10.1016/S0022-2275(20)37981-5
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      Rabbits were fed either 10% coconut oil, 10% coconut oil and 1% beta-sitosterol, 10% coconut oil and 1% cholesterol, or 10% coconut oil and 1% beta-sitosterol plus 1% cholesterol for 4 weeks. Microsomal membranes from intestines of animals fed the 1% beta-sitosterol diet had 48% less cholesterol and were enriched twofold in beta-sitosterol compared to membranes from animals fed the coconut oil diet alone. Acylcoenzyme A:cholesterol acyltransferase (ACAT) activity in jejunum and ileum was decreased significantly in animals fed the plant sterol alone. In membranes from animals fed 1% beta-sitosterol and 1% cholesterol, beta-sitosterol content increased 50% whereas cholesterol was modestly decreased compared to their controls fed only cholesterol. Intestinal ACAT was unchanged in the animals fed both sterols when compared to their controls. beta-Sitosterol esterification was determined by incubating intestinal microsomal membranes with either [(14)C]beta-sitosterol-albumin emulsion or [(14)C]beta-sitosterol:dipalmitoyl phosphatidylcholine (DPPC) liposomes to radiolabel the endogenous sterol pool. Oleoyl-CoA was then added. The CoA-dependent esterification rate of beta-sitosterol was very slow compared to that of cholesterol using both techniques. An increased amount of endogenous microsomal beta-sitosterol, which occurs in animals fed 1% beta-sitosterol, did not interfere with the stimulation of ACAT activity secondary to cholesterol enrichment of the membranes. Enriching microsomal membranes three- to five-fold with beta-sitosterol did not affect ACAT activity. Freshly isolated intestinal cells were incubated for 1 hour with [(3)H]oleic acid and beta-sitosterol:DPPC or 25-hydroxycholesterol:DPPC. Incorporation of oleic acid into cholesteryl esters did not change in the presence of beta-sitosterol but increased fourfold after the addition of 25-hydroxycholesterol. We conclude that the CoA-dependent esterification rate of cholesterol is at least 60 times greater than that of beta-sitosterol. Membrane beta-sitosterol does not interfere with nor compete with cholesterol esterification. Inadequate esterification of this plant sterol may play a role in the poor absorption of beta-sitosterol by the gut.-Field, F. J., and S. N. Mathur. beta-Sitosterol: esterification by intestinal acylcoenzyme A:cholesterol acyltransferase (ACAT) and its effect on cholesterol esterification.

      REFERENCES

        • Gould R.G.
        Absorbability of ß-sitosterol.
        Trans. NY Acad. Sci. 1955; 18: 129-134
        • Gould R.G.
        • Jones R.J.
        • LeRoy G.V.
        • Wissler R.W.
        • Taylor C.B.
        Absorbability of ß-sitosterol in humans.
        Metabolism. 1969; 18: 652-662
        • Grundy S.M.
        • Ahrens Jr., E.H.
        • Salen G.
        Dietary ß-sitosterol as an internal standard to correct for cholesterol losses in sterol balance studies.
        J. Lipid Res. 1968; 9: 374-387
        • Borgström B
        Quantification of cholesterol absorption in man by fecal analysis after the feeding of a single isotope-labeled meal.
        J. Lipid Res. 1969; 10: 331-337
        • Salen G.
        • Ahrens Jr., E.H.
        • Grundy S.M.
        Metabolism of ß-sitosterol in man.
        J. Clin. Invest. 1970; 49: 952-967
        • Samuel P.
        • Crouse J.R.
        • Ahrens Jr., E.H.
        Evaluation of an isotope ratio method for measurement of cholesterol absorption in man.
        J. Lipid Res. 1978; 19: 82-93
        • McNamara D.J.
        • Davidson N.O.
        • Samuel P.
        • Ahrens Jr., E.H.
        Cholesterol absorption in man: effect of administration of clofibrate and/or cholestyramine.
        J. Lipid Res. 1980; 21: 1058-1064
        • Mok H.Y.I.
        • von Bergmann K.
        • Grundy S.M.
        Effects of continuous and intermittent feeding on biliary lipid outputs in man: application for measurements of intestinal absorption of cholesterol and bile acids.
        J. Lipid Res. 1979; 20: 389-398
        • Grundy S.M.
        • Mok H.Y.I.
        Determination of cholesterol absorption in man by intestinal perfusion.
        J. Lipid Res. 1977; 18: 263-271
        • Connor W.E.
        • Lin D.
        The intestinal absorption of dietary cholesterol by hypercholesterolemic (type II) and normocholesterolemic humans.
        J. Clin. Invest. 1974; 53: 1062-1070
        • Swell L.
        • Trout Jr., E.C.
        • Field Jr., H.
        • Treadwell C.R.
        Absorption of 3H-β-sitosterol in the lymph fistula rat.
        Proc. Soc. Exp. Biol. Med. 1959; 100: 140-142
        • Sylvén C.
        • Borgström B.
        Absorption and lymphatic transport of cholesterol and sitosterol in the rat.
        J. Lipid Res. 1969; 10: 179-182
        • Borgström B
        Quantitative aspects of the intestinal absorption and metabolism of cholesterol and ß-sitosterol in the rat.
        J. Lipid Res. 1968; 9: 473-481
        • Kuksis A.
        • Huang T.C.
        Differential absorption of plant sterols in the dog.
        Can. J. Biochem. Physiol. 1962; 40: 1493-1504
        • Bhattacharyya A.K.
        In vitro esterification of plant sterols by the esterifying enzyme of the small intestine of rat.
        Experientia. 1979; 35: 1614-1615
        • Bhattacharyya A.K.
        • Connor W.E.
        β-Sitosterolemia and xanthomatosis.
        J. Clin. Invest. 1974; 53: 1033-1043
        • Farquhar J.W.
        • Smith R.E.
        • Dempsy M.E.
        The effect of β-sitosterol on the serum lipids of young men with arteriosclerotic heart disease.
        Circulation. 1956; 14: 77-82
        • Grundy S.M.
        • Ahrens Jr., E.H.
        • Davignon J.
        The interaction of cholesterol absorption and cholesterol synthesis in man.
        J. Lipid Res. 1969; 10: 304-315
        • Lees A.M.
        • Mok H.Y.I.
        • Lees R.S.
        • McCluskey M.A.
        • Grundy S.M.
        Plant sterols as cholesterol-lowering agents: clinical trials in patients with hypercholesterolemia and studies of sterol balance.
        Atherosclerosis. 1977; 28: 325-338
        • Grundy S.M.
        • Mok H.Y.I.
        Colestipol, clofibrate, and phytosterols in combined therapy of hyperlipidemia.
        J. Lab. Clin. Med. 1977; 89: 354-366
        • Helgerud P.
        • Saarem K.
        • Norum K.R.
        Acyl-CoAxholesterol acyltransferase in human small intestine: its activity and some properties of the enzymic reaction.
        J. Lipid Res. 1981; 22: 271-277
        • Field F.J.
        • Cooper A.D.
        • Erickson S.K.
        Regulation of rabbit intestinal acyl coenzyme A-cholesterol acyltransferase in vivo and in vitro.
        Gastroenterology. 1982; 83: 873-880
        • Field J.
        • Salome R.
        Effect of dietary fat saturation and cholesterol on intestinal acylcoenzyme Axholesterol acyltransferase (ACAT) activity.
        Biochim. Biophys. Acta. 1982; 712: 557-570
        • Haugen R.
        • Norum K.R.
        Coenzyme-A-dependent esterification of cholesterol in rat intestinal mucosa.
        Scand. J. Gastroenterol. 1976; 11: 615-621
        • Stokke K.T.
        • Norum K.R.
        Determination of lecithin: cholesterol acyltransferase in human blood plasma.
        Scand. J. Clin. Lab. Invest. 1971; 27: 21-27
        • Erickson S.K.
        • Shrewsbury M.A.
        • Brooks C.
        • Meyer D.J.
        Rat liver acyl-coenzyme A:cholesterol acyltransferase: its regulation in vivo and some of its properties in vitro.
        J. Lipid Res. 1980; 21: 930-941
        • Raheja R.K.
        • Kaur C.
        • Singh A.
        • Bhatia I.S.
        New colorimetric method for the quantitative estimation of phospholipids without acid digestion.
        J. Lipid Res. 1973; 14: 695-697
        • Lowry O.H.
        • Rosebrough N.J.
        • Farr A.L.
        • Randall R.J.
        Protein measurement with the Folin phenol reagent.
        J. Biol. Chem. 1951; 193: 265-275
        • Weiser M.M.
        Intestinal epithelial cell surface membrane glycoprotein synthesis.
        J. Biol. Chem. 1973; 248: 2536-2541
        • Drevon C.A.
        • Weinstein D.B.
        • Steinberg D.
        Regulation of cholesterol esterification and biosynthesis in monolayer cultures of normal adult rat hepatocytes.
        J. Biol. Chem. 1980; 255: 9128-9137
        • Tavani D.M.
        • Nes W.R.
        • Billheimer J.T.
        The sterol substrate specificity of acyl CoA:cholesterol acyltransferase from rat liver.
        J. Lipid Res. 1982; 23: 774-781
        • Borgström B
        Partition of lipids between emulsified oil and micellar phases of glyceride-bile salt dispersions.
        J. Lipid Res. 1967; 8: 598-608
        • Brown M.S.
        • Ho Y.K.
        • Goldstein J.L.
        The cholesteryl ester cycle in macrophage foam cells.
        J. Biol. Chem. 1980; 255: 9344-9352
        • Goldstein J.L.
        • Dana S.E.
        • Brown M.S.
        Esterification of low density lipopotein cholesterol in human fibroblasts and its absence in homozygous familial hypercholesterolemia.
        Proc. Natl. Acad. Sci. USA. 1974; 71: 4288-4292
        • Nilsson A
        Increased cholesterol-ester formation during forced cholesterol synthesis in rat hepatocytes.
        Eur. J. Biochem. 1975; 51: 337-342