, Volume 38, Issue 12, pp 1237–1247 | Cite as

Comparative health effects of margarines fortified with plant sterols and stanols on a rat model for hemorrhagic stroke

  • W. M. N. Ratnayake
  • L. Plouffe
  • M. R. L’Abbé
  • K. Trick
  • R. Mueller
  • S. Hayward


There is increased acceptance of fortifying habitual foods with plant sterols and their saturated derivatives, stanols, at levels that are considered safe. These sterols and stanols are recognized as potentially effective dietary components for lowering plasma total and LDL cholesterol. Our previous studies have shown that daily consumption of plant sterols promotes strokes and shortens the life span of stroke-prone spontaneously hypertensive (SHRSP) rats. These studies question the safety of plant sterol additives. The present study was performed to determine whether a large intake of plant stanols would cause nutritional effects similar to those seen with plant sterols in SHRSP rats. Young SHRSP rats (aged 26–29 d) were fed semipurified diets containing commercial margarines fortified with either plant stanols (1.1 g/100 g diet) or plant sterols (1.4 g/100 g diet). A reference group of SHRSP rats was fed a soybean oil diet (0.02 g plant sterols/100 g diet and no plant stanols). Compared to soybean oil, both plant stanol and plant sterol margarines significantly (P<0.05) reduced the life span of SHRSP rats. At the initial stages of feeding, there was no difference in the survival rates between the two margarine groups, but after approximately 50 d of feeding, the plant stanol group had a slightly, but significantly (P<0.05), lower survival rate. Blood and tissue (plasma, red blood cells, liver, and kidney) concentrations of plant sterols in the plant sterol margarine group were three to four times higher than the corresponding tissue concentrations of plant stanols in the plant stanol group. The deformability of red blood cells and the platelet count of SHRSP rats fed, the plant sterol margarine were significantly (P<0.05) lower than those of the plant stanol margarine and soybean oil groups at the end of the study. These parameters did not differ between the soybean oil and plant stanol margarine groups. These results suggest that, at the levels tested in the present study, plant stanols provoke hemorrhagic stroke in SHRSP rats to a slightly greater extent than plant sterols. The results also suggest that the mechanism by which plant stanols shorten the life span of SHRSP rat might differ from that of plant sterols.



ATP transporter binding cassette


American Institute of Nutrition


deformability index


laser-assisted optical rotational cell analyzer


cis-monounsaturated FA




red blood cell

SHRSP rats

stroke-prone spontaneously hypertensive rats


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  1. 1.
    Gylling, H., and Miettinen, T.A. (1994) Serum Cholesterol and Lipoprotein Metabolism in Hypercholesterolemic NIDDM Patients Before and During Sitostanol Ester Margarine Treatment, Diabetologia 37, 773–780.PubMedGoogle Scholar
  2. 2.
    Miettinen, T.A., Puska, P., Gyling, H., Vanhanen, H., and Vartiainen, E. (1995) Reduction of Serum Cholesterol with Sitostanol-Ester Margarine in a Mildly Hypercholesterolemic Population, New Engl. J. Med. 333, 1308–1312.PubMedCrossRefGoogle Scholar
  3. 3.
    Gylling, H., Siimes, M.A., and Miettinen, T.A. (1995) Sitostanol Ester Margarine in Dietary Treatment of Children with Familial Hypercholesterolemia, J. Lipid Res. 36, 1807–1812.PubMedGoogle Scholar
  4. 4.
    Gylling, H., Radhakrishnan, R., and Miettinen, T.A. (1997) Reduction of Serum Cholesterol in Postmenopausal Women with Previous Myocardial Infarction and Cholesterol Malabsorption Induced by Dietary Sitostanol Margarine. Women and Dietary Sitostanol, Circulation 96, 4226–4231.PubMedGoogle Scholar
  5. 5.
    Westrate, J.A., and Meijer, G.W. (1998) Plant Sterol-Enriched Margarines and Reduction of Plasma Total- and LDL-Cholesterol Concentrations in Normocholesterolaemic and Mildly Hypercholesterolaemic Subjects, Eur. J. Clin. Nutr. 52, 334–343.CrossRefGoogle Scholar
  6. 6.
    Plat, J., and Mensink, R.P. (2001) Effects of Plant Sterols and Stanols on Lipid Metabolism and Cardiovascular Risk, Nutr. Metabol. Cardiovasc. Dis. 11, 31–40.Google Scholar
  7. 7.
    Tammi, A., Ronnemaa, T., Gylling, H., Rask-Nissila, L., Viikari, J., Tuominen, J., Pulkki, K., and Simell, O. (2002) Plant Stanol Ester Margarine Lowers Serum Total and Low-Density Lipoprotein Cholesterol Concentrations of Healthy Children: The STRIP Project. Special Turku Coronary Risk Factors Intervention Project, J. Pediatr. 136, 503–510.Google Scholar
  8. 8.
    de Graff, J., de Sauvage Nolting, P.R.W., van Dam, M., Belsey, E.M., Kastelein, J.J.P., Pritchard, P.H., and Stalenhoef, A.F.H. (2002) Consumption of Tall Oil-Derived Phytosterols in a Chocolate Matrix Significantly Decreases Plasma Total and Low-Density Lipoprotein-Cholesterol Levels, Br. J. Nutr. 88, 479–488.CrossRefGoogle Scholar
  9. 9.
    Miettinen, T.A., Tilvis, R.S., and Kesaniemi, Y.A. (1990) Serum Plant Sterols and Cholesterol Precursors Reflect Cholesterol Absorption and Synthesis in Volunteers of a Randomly Selected Male Population, Am. J. Epidemiol. 131, 20–31.PubMedGoogle Scholar
  10. 10.
    Cater, N.B., and Grundy, S.M. (1998) Safety Aspects of Dietary Plant Sterols and Stanols: Managing High Cholesterol, in A Postgraduate Medicine Special Report (Roberts, W.O., ed.), pp. 6–14, McGraw-Hill, Minneapolis.Google Scholar
  11. 11.
    Law, M. (2000) Plant Sterol and Stanol Margarines and Health, Br. Med. J. 320, 861–864.CrossRefGoogle Scholar
  12. 12.
    Ling, W.H., and Jones, P.J.H. (1995) Dietary Phytosterols: A Review of Metabolism, Benefits and Side Effects, Life Sci. 157, 196–206.Google Scholar
  13. 13.
    Moghadasian, M.H., and Frohlich, J.J. (1999) Effects of Dietary Phytosterols on Cholesterol Metabolism and Atherosclerosis: Clinical and Experimental Evidence, Am. J. Med. 107, 588–594.PubMedCrossRefGoogle Scholar
  14. 14.
    Ratnayake W.M.N., and Vavasour, E., (2003) Potential Health Risks Associated with Large Intakes of Plant Sterols, in Phytosterols: as Functional Food Components and Nutraceuticals (Dutta, P., ed.), pp. 365–394, Marcel Dekker, New York.Google Scholar
  15. 15.
    Björkhem, I., Boberg, K.M., and Leitersdorf, E. (2001) Inborn Errors in Bile Acid Biosynthesis and Storage of Sterols Other than Cholesterol, in The Metabolic and Molecular Bases of Inherited Disease (Scriver, C.R., Beaudet, A.L., Sly, W.S., and Valle, D., eds.), pp. 2961–2988, McGraw-Hill, New York.Google Scholar
  16. 16.
    Iyer, K.R., Spitz, L., and Clayton, P. (1998) New Insight into Mechanisms of Parental Nutrition-Associated Cholestasis: Role of Plant Sterols, J. Pediatr. Surg. 33: 1–6.PubMedCrossRefGoogle Scholar
  17. 17.
    Glueck, C.J., Spiers, J., Tracy, T., Streicher, P., Illig, E., and Vandegrift, J. (1991) Relationships of Serum Plant Sterols (Phytosterols) and Cholesterol in 595 Hypercholesterolemic Subjects, and Familial Aggregation of Phytosterols, Cholesterol, and Premature Coronary Heart Disease in Hyperphytosterolemic Probands and Their First Degree Relatives, Metabolism 40, 842–848.PubMedCrossRefGoogle Scholar
  18. 18.
    Sutherland, W.H.F., Williams, M.J.A., Nye, E.R., Restieaux, N.J., de Jong, S.A., and Walker, H.J. (1998) Associations of Plasma Noncholesterol Sterol Levels with Severity of Coronary Artery Disease, Nutr. Metabol. Cardiovasc. Dis. 8, 386–391.Google Scholar
  19. 19.
    Rajaratnam, R.A., Gylling, H., and Miettinen, T.A. (2000) Independent Association of Serum Squalene and Noncholesterol Sterols with Coronary Artery Disease in Postmenopausal Women, J. Am. Coll. Cardiol. 35, 1185–1191.PubMedCrossRefGoogle Scholar
  20. 20.
    Sudhop, T., Gottwald, B.M., and Bergman, K. (2002) Serum Plant Sterols as a Potential Risk Factor for Coronary Heart Disease, Metabolism 12, 1519–1521.CrossRefGoogle Scholar
  21. 21.
    Ratnayake, W.M.N., L’Abbé, M.R., Mueller, R., Hayward, S., Plouffe, L., Hollywood, R., and Trick, K. (2000) Vegetable Oils high in Phytosterols Make Erythrocytes Less Deformable and Shorten the Life Span of Stroke-Prone Spontaneously Hypertensive Rats, J. Nutr. 130, 1166–1178.PubMedGoogle Scholar
  22. 22.
    Ratnayake, W.M.N., Plouffe, L., Hollywood, R., L’Abbé, M.R., Hidiroglou, N., Sarwar, G., and Mueller, R. (2000) Influence of Source of Dietary Oils on the Life Span of Stroke-Prone Spontaneously Hypertensive Rats, Lipids 35, 409–420.PubMedCrossRefGoogle Scholar
  23. 23.
    Yamori, Y., Nara, Y., Mizushima, S., Murakami, S., Ikeda, K., Sawamura, M., Nabika, T., and Horie, R. (1992) Gene-Environment Interaction in Hypertension, Stroke and Atherosclerosis in Experiment Models and Supportive Findings from a World-wide Cross-Sectional Epidemiological Survey: A WHO-Cardiac Study, Clin. Exp. Pharmacol. Physiol. 19, 43–52.Google Scholar
  24. 24.
    Heinemann, T., Axtmann, G., and von Bergmann, K. (1993) Comparison of Intestinal Absorption of Cholesterol with Different Plant Sterols in Man, Eur. J. Clin. Invest. 23, 827–831.PubMedCrossRefGoogle Scholar
  25. 25.
    Sanders, D.J., Minter, H.J., Howes, D., and Hepburn, P.A. (2000) The Safety Evaluation of Phytosterol Esters. Part 6. The Comparative Absorption and Tissue Distribution of Phytosterols in the Rat, Food Chem. Toxicol. 38, 485–491.PubMedCrossRefGoogle Scholar
  26. 26.
    Lawless, J.F. (1982) Statistical Models and Methods for Lifetime Data, John Wiley & Sons, New York.Google Scholar
  27. 27.
    Reeves, P.G., Nielsen, F.H., and Fahey, G.C. (1993) AIN-93 Purified Diets for Laboratory Rodents: Final Report of the American Institute of Nutrition ad hoc Committee on the Reformulation of the AIN-76A Rodent Diet, J. Nutr. 123, 1939–1951.PubMedGoogle Scholar
  28. 28.
    Wood, D., De Backer, G., Faergeman O., Graham, I., Mancia, G., and Pyörälä, K., together with members of the Task Force (1998) Prevention of Coronary Heart Disease in Clinical Practice. Recommendations of the Second Joint Task Force of European and Other Societies on Coronary Prevention, Eur. Heart J. 19, 1434–1503.CrossRefGoogle Scholar
  29. 29.
    Krauss, R.M., Eckel, R.H., Howard, B., Appel, L.J., Daniels, S.R., Deckelbaum, R.J., Erdman, J.W., Kris-Etherton, P., Goldberg, I.J., Kotchen, T.A., et al. (2000) AHA Dietary Guidelines. Revision 2000: A Statement for Healthcare Professionals from the Nutrition Committee of the American Heart Association, Circulation 102, 2284–2299.PubMedGoogle Scholar
  30. 30.
    Report of the Panel on Macronutrients of the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes (2002) Dietary Reference Intakes for Energy, Carbohydrates, Fiber, Fat, Fatty Acids, Protein, and Amino Acids, pp. 8-1–8-97. Food and Nutrition Board, Institute of Medicine, The National Academies Press, Washington, DC.Google Scholar
  31. 31.
    Yamori, Y. (1983) Physiology of the Various Strains of Spontaneously Hypertensive Rats, in Hypertension (Genest, J., Kuchel, O., Hamet, P., and Cantin, M., eds.), pp. 556–581, McGraw-Hill, Montreal.Google Scholar
  32. 32.
    Yamori, Y., (1984) The Stroke-Prone Spontaneously Hypertensive Rat: Contribution to Risk Factor Analysis and Prevention of Diseases, in Handbook of Hypertension (de Jong, W., ed.), Vol. 4: Experimental and Genetic Models of Hypertension, pp. 240–255, Elsevier Science, Amsterdam.Google Scholar
  33. 33.
    Yamori, Y. (1989) Predictive and Preventive Pathology of Cardiovascular Diseases, Acta Pathol. Jpn. 39, 683–705.PubMedGoogle Scholar
  34. 34.
    Huang, M.Z., Naito, Y., Watanabe, S., Kobayashi, T., Kanai, H., Nagai, H., and Okuyama, H. (1996) Effect of Rapeseed and Dietary Oils on the Mean Survival Time of Stroke-Prone Spontaneously Hypertensive Rats, Biol. Pharmacol. Bull. 19, 554–557.Google Scholar
  35. 35.
    Huang, M.Z., Watanabe, S., Kobayashi, T., Nagatsu, A., Sakakibara, J., and Okuyama, H. (1997) Unusual Effects of Some Vegetable Oils on the Survival Time of Stroke-Prone Spontaneously Hypertensive Rats, Lipids 32, 745–751.PubMedCrossRefGoogle Scholar
  36. 36.
    Hobbs, L.M., Rayner, T.E., and Howe, P.R.C. (1996) Dietary Fish Oil Prevents the Development of Renal Damage in Salt-Loaded Stroke-Prone Spontaneously Hypertensive Rats, Clin. Exp. Pharmacol. Physiol. 23, 508–513.PubMedGoogle Scholar
  37. 37.
    Iso, H., Stampfer, M.J., Manson, J.E., Rexrode, K., Hu, F.B., Hennekens, C.H., Colditz, G.A., Speizer, F.E., and Willett, W.C. (2001) Prospective Study of Fat and Protein Intake and Risk of Intraparenchymal Hemorrhage in Women, Circulation 103, 856–863.PubMedGoogle Scholar
  38. 38.
    Komachi, Y., Iida, M., and Ozawa, M. (1977) Risk Factors for Stroke, Saisha Igaku 32, 2264–2269.Google Scholar
  39. 39.
    Ueshima, H., Iida, M., Shimamoto, T., Konishi, M., Tsujioka, K., Tanigaki, M., Nakanishi, N., Ozawa, H., Kojima, S., and Komachi, Y. (1980) Multivariate Analysis of Risk Factor for Stroke: Eight-Year Follow-up Study of Farming Villages in Akita Japan, Prev. Med. 9, 722–740.PubMedCrossRefGoogle Scholar
  40. 40.
    Tanaka, H., Ueda, Y., Hayashi, M., Date, C., Baba, T., Yamashita H., Shoji, H., Tanaka, Y., Owada, K., and Detels, R. (1982) Risk Factors for Cerebral Hemorrhage and Cerebral Infarction in a Japanese Rural Community, Stroke 13, 62–73.PubMedGoogle Scholar
  41. 41.
    Yano, K., Reed, D.M., and MacLean, C.J. (1989) Serum Cholesterol and Hemorrhagic Stroke in the Honolulu Heart Program, Stroke 20, 1460–1465.PubMedGoogle Scholar
  42. 42.
    Iso, H., Jacobs, D.R., Wentworth, D., Neaton, J.D., and Cohen, J.D. (1989) Serum Cholesterol Levels and Six-Year Mortality from Stroke in 350,977 Men Screened for the Multiple Risk Factor Intervention Trial, N. Engl. J. Med. 320, 904–910.PubMedCrossRefGoogle Scholar
  43. 43.
    Miyazaki, M., Huang, M.-Z., Watanabe, S., Kobayashi, T., and Okuyama, H. (1998) Early Mortality Effect of Partially Hydrogenated Vegetable Oils in Stroke-Prone Spontaneously Hypertensive Rats (SHRSP), Nut. Res. 18, 1049–1056.CrossRefGoogle Scholar
  44. 44.
    Ackman, R.G., Hooper, S.N., and Hooper, D.L. (1974) Linolenic acid Artifacts from the Deodorization of Oils, J. Am. Oil Chem. Soc. 51, 42–49.Google Scholar
  45. 45.
    Hamano, M., Mashiko, S., Onda, T., Tomita, I., and Tomita, T. (1995) Effects of Cholesterol-Diet on the Incidence of Stroke and Life Span in Malignant Stroke-Prone Spontaneously Hypertensive Rats, Jpn. Heart J. 36, 511.Google Scholar
  46. 46.
    Sugano, M., Kamo, F., Ikeda, I., and Morioka, H. (1976) Lipid-Lowering Activity of Phytostanols in Rats, Atherosclerosis 24, 301–309.PubMedCrossRefGoogle Scholar
  47. 47.
    Sugano, M., Morioka, H., and Ikeda, I. (1977) A Comparison of Hypocholesterolemic Activity of β-Sitosterol and β-Sitostanol in Rats, J. Nutr. 107, 2011–2019.PubMedGoogle Scholar
  48. 48.
    Ikeda, I., and Sugano, M. (1978) Comparison of Absorption and Metabolism of β-Sitosterol and β-Sitostanol in Rats, Atherosclerosis 30, 227–237.PubMedCrossRefGoogle Scholar
  49. 49.
    Westrate, J.A., and Meijer, G.W. (1998) Plant Sterol-Enriched Margarine and Reduction of Plasma Total- and LDL-Cholesterol Concentrations in Normocholesterolemic and Mildly Hypercholesterolemic Subjects, Eur. J. Clin. Nutr. 52, 334–343.CrossRefGoogle Scholar
  50. 50.
    Jones, P.J.H., Raeini-Sarjaz, M., Ntanios, F.Y., Vanstone, C.A., Feng, J.Y., and Parsons, W.E. (2000) Modulation of Plasma Lipid Levels and Cholesterol Kinetics by Phytosterol Versus Phytostanol Esters, J. Lipid Res. 41, 697–705.PubMedGoogle Scholar
  51. 51.
    Vanstone, C.A., Raeini-Sarjaz, M., Parsons, W.E., and Jones, P.J.H. (2002) Unesterified Plant Sterols and Stanols Lower LDL-Cholesterol Concentrations Equivalently in Hypercholesterolemic Persons, Am. J. Clin. Nutr. 76, 1272–1278.PubMedGoogle Scholar
  52. 52.
    Heinemann, T., Leiss, O., and von Bergmann, K. (1986) Effect of Low-Dose Sitostanol on Serum Cholesterol in Patients with Hypercholesterolemia, Atherosclerosis 61, 219–223.PubMedCrossRefGoogle Scholar
  53. 53.
    Miettinen, T.A., Puska, P., Gylling, H., Vanhanen, H.T., and Vartianen, E. (1995) Reduction of Serum Cholesterol with Sitostanol-Ester Margarine in a Mildly Hypercholesterolemic Population, N. Engl. J. Med. 333, 1308–1312.PubMedCrossRefGoogle Scholar
  54. 54.
    Heinemann, T., Kullak-Ublick, G.-A., Pietruck, B., and von Bergmann, K. (1991) Mechanisms of Action of Plant Sterols on Inhibition of Cholesterol Absorption, Comparison of Sitosterol and Sitostanol, Eur. J. Pharmacol. 40, S59-S63.CrossRefGoogle Scholar
  55. 55.
    Becker, M., Staab, D., and von Bergmann, K. (1993) Treatment of Severe Familial Hypercholesterolemia in Childhood with Sitosterol and Sitostanol, J. Pediatr. 122, 292–296.PubMedGoogle Scholar
  56. 56.
    Gylling, H., and Miettinen, T.A. (1999) Cholesterol Reduction by Different Plant Sterol Mixtures and with Variable Fat Intake, Metabolism 48, 575–580.PubMedCrossRefGoogle Scholar
  57. 57.
    Naito, Y., Konishi, C., Katsumara, H., and Ohara, N. (2000) Increase in Blood Pressure with Enhanced Na+, K+-ATPase Activity in Stroke-Prone Spontaneously Hypertensive Rats After 4-Week’s Intake of Rapeseed Oil as the Sole Dietary Fat, Pharmacol. Toxicol. 87, 144–148.PubMedCrossRefGoogle Scholar
  58. 58.
    Naito, Y., Nagata, T., Takano, Y., and Ohara, N. (2003) Rape-seed Oil Ingestion and Exacerbation of Hypertension Related Conditions in Stroke-Prone Spontaneously Hypertensive Rats, Toxicology 187, 144–148.CrossRefGoogle Scholar
  59. 59.
    Naito, Y., Konishi, C., and Ohara, N. (2000) Blood Coagulation and Osmotic Tolerance of Erythrocytes in Stroke-Prone Hypertensive Rats Given Rapeseed Oil as the Only Dietary Fat, Toxicol. Lett. 117, 209–215.CrossRefGoogle Scholar
  60. 60.
    Naito, Y., Yoshida, H., Nagata, T., Tanaka, A., Ono, H., and Ohara, N. (2000) Dietary Intake of Rapeseed Oil or Soybean Oil as the Only Fat Nutrient in spontaneously Hypertensive Rats and Wistar Kyoto Rats—Blood Pressure and Pathophysiology, Toxicology 146, 197–208.PubMedCrossRefGoogle Scholar
  61. 61.
    Scoggan, K.A., Gruber, H., and Larivière, K. (2003) A Missence Mutation in the Rat Abeg5 Gene Causes Phytosterolemia in Stroke-Prone Spontaneously Hypertensive Rats (SHRSP), Spontaneously Hypertensive and Normotensive (WKY Inbred) Rats, J. Lipid Res. 44, 911–916.PubMedCrossRefGoogle Scholar
  62. 62.
    Berge, M.H., Tian, H., Graf, G.A., Yu, L., Grishin, N.V., Schultz, J., Kwiterovich, P., Shan, B., Barnes, R., and Hobbs, H.H. (2000) Accumulation of Dietary Cholesterol in Sitosterolemia Caused by Mutations in Adjacent ABC Transporters, Science 290, 1771–1775.PubMedCrossRefGoogle Scholar
  63. 63.
    Lee, M.H., Lu, K., Hazard, S., Yu, H., Shulenin, S., Hidaka, H., Kojima, H., Allikmets, R., Sakuma, N., Pegoraro, R., et al. (2001) Identification of a Gene, ABCG5, Important in the Regulation of Dietary Cholesterol Absorption, Nat. Genet. 27, 79–83.PubMedCrossRefGoogle Scholar
  64. 64.
    Hubacek, J.A., Berge, K.E., Cohen, J.C., and Hobbs, H.H. (2001) Mutations in ATP-Cassette Binding Proteins G5 (ABCG5) and G8 (ABCG8) Causing Sitosterolemia, Hum. Mutat. 18, 359–360.PubMedCrossRefGoogle Scholar
  65. 65.
    Heimer, S., Langman, T., Moehle, C., Mauerer, R., Dean, M., Beil, F.U., von Bergmann, K., and Schmitz, G. (2002) Mutations in the Human ATP-Binding Cassette Transporters ABCG5 and ABCG8 in Sitosterolemia, Hum. Mutat. 20, 151.CrossRefGoogle Scholar

Copyright information

© AOCS Press 2003

Authors and Affiliations

  • W. M. N. Ratnayake
    • 3
  • L. Plouffe
    • 3
  • M. R. L’Abbé
    • 3
  • K. Trick
    • 3
  • R. Mueller
    • 1
  • S. Hayward
    • 2
  1. 1.Toxicology Research DivisionFood Directorate, Health Products and Food Branch, Health CanadaOttawaCanada
  2. 2.Bureau of Biostatistics and Computer ApplicationsFood Directorate, Health Products and Food Branch, Health CanadaOttawaCanada
  3. 3.Nutrition Research Division, Food Directorate, Health Products and Food BranchHealth CanadaOttawaCanada

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