Skip to main content
SpringerLink
Log in

Regulation of the apolipoprotein E by dietary lipids occurs by transcriptional and post-transcriptional mechanisms

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The aim of the present investigation was to study the regulation of apolipoprotein E by two dietary nutrients, saturated fat and cholesterol, known to raise plasma cholesterol levels. ApoE is a protein component of several classes of lipoproteins including VLDL and HDL, and dietary lipids may regulate VLDL and apoE-containing HDL particles through their effects on apoE gene. Male rats and mice were fed the following 4 diets: control diet (C); high cholesterol diet with 0.5% cholesterol (HC); high fat diet with 20% hydrogenated coconut oil (HF); and high fat plus high cholesterol diet with 0.5% cholesterol and 20% fat (HF/C). Plasma cholesterol levels remained unchanged on HC diet, but in mice VLDL-cholesterol increased by 31%. HF diet increased VLDL and LDL by 15–17% in rats, and 21% in mice. A combination of fat and cholesterol diet showed pronounced effects on plasma lipoprotein concentrations, raising apoB-containing particles by 21% and 44% in mice and rats, respectively. Plasma apoE levels increased significantly on all diets. The mechanism of regulation of increased plasma apoB and apoE levels was examined. Quantification of hepatic apoB mRNA showed a lack of correlation between plasma apoB and hepatic apoB mRNA levels, suggesting that posttranscriptional regulation increased plasma apoB-containing lipoproteins in animals fed saturated fat diets. Hepatic apoE mRNA levels increased significantly in animals fed cholesterol-rich diets. However, despite increased plasma apoE levels on diet containing only saturated fat, hepatic apoE mRNA did not change. Synthesis of apoE on the liver polysomes increased selectively on cholesterol-rich diets. These results suggest that cholesterol-rich diets altered apoE, in part, by transcriptional mechanism, and saturated fat-rich diets increased plasma apoE levels by posttranscriptional mechanism, possibly decreased receptor-mediated uptake of apoE-containing particles. The regulation of LDL receptor was also studied since plasma apoB and E levels may be altered by LDL receptor-mediated uptake by the hepatocytes. As expected, high cholesterol diet decreased LDL receptor mRNA by 30–40%. However, the LDL receptor protein on liver membranes did not change on any of the test diets in both animal species. Hepatic cholesterol content increased several fold selectively on high cholesterol diets. These findings suggest that: 1) both transcriptional and posttranscriptional mechanisms are important in regulating plasma apoB and E containing lipoproteins; 2) dietary cholesterol regulates apoE gene by a transcriptional mechanism anddietary saturated fat by posttranscriptional mechanism; and 3) changes in the hepatic apoE and LDL receptor mRNA are associated with the changes in intracellular cholesterol concentrations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

VLDL:

very low density lipoprotein

LDL:

low density lipoprotein

HDL:

high density lipoprotein; apoapolipoprotein

EDTA:

ethylene diamine tetra-acetic acid

SDS:

sodium dodecyl sulfate

HC:

high cholesterol

HF:

high fat

HF/C:

high fat plus cholesterol

References

  1. Goldberg A, Schonfeld G: Effects of diets on lipoprotein metabolism. Ann Rev Nutr 5: 195–21, 1985

    Google Scholar 

  2. Miller GJ, Miller NE: Plasma high density lipoproteins and the development of ischaemic heart diseases. Lancet 1: 16–19, 1975

    Article  CAS  PubMed  Google Scholar 

  3. Rall SC, Weisgraber KH, Mahley RW: Abnormal lipoprotein receptor binding activity of the human apoE apoprotein due to cysteine-arginine interchange at a single site. J Biol Chem 257: 2518–2521, 1982

    Google Scholar 

  4. Mahley RW: Apolipoprotein E: cholesterol transport protein with expanding role in cell biology. Science 240: 622–630, 1988

    Google Scholar 

  5. Ishibashi S, Herz J, Maeda N, Goldstein JL, Brown MS: The two receptor model of lipoprotein clearance: Tests of the hypothesis in “knockout” mice lacking the low density lipoprotein receptor, apolipoprotein E, or both proteins. Proc Natl Acad Sci USA 91: 443l-4435, 1994

    Google Scholar 

  6. Innerarity TL, Mahley RW: Enhanced binding of cultured human fibroblasts of apoE-containing lipoproteins as compared with low density lipoproteins. Biochemistry 17: 1440–1447, 1978

    Google Scholar 

  7. Shimano H, Yamada N, Katsuki M, Shimada M, Gotoda T, Harada K, Murase T, Fukazawa C, Takaku F, Yazaka Y: Overexpression of apolipoprotein E in transgenic mice: Marked reduction in plasma lipoproteins except high density lipoproteins and resistance against diet-induced hypercholesterolemia. Proc Natl Acad Sci USA 89: 1750–1754, 1992

    Google Scholar 

  8. Staels B, Auwrex J, Chan L, van Tol A, Rosseneu M, Verhoeven G: Influence of development, estrogen, and food intake on apolipoprotein AI, All and E mRNA in rat liver and intestine. J Lipid Res 30: 1137–1145, 1989

    Google Scholar 

  9. Lin-Lee Y-C, Tanaka Y, Lin C-T, Chan L: Effects of atherogenic diet on apolipoprotein E biosynthesis in the rat. Biochemistry 20: 6474–6480, 1981

    Google Scholar 

  10. Mazzone T, Basheeruddin K, Paulos C: Regulation of macrophage apolipoprotein E gene expression by cholesterol. J Lipid Res 30: 1055–1064, 1989

    Google Scholar 

  11. Strobl W, Gorder NL, Fienup GA, Lin-Lee Y-C, Gotto AM Jr, Patsch W: Effect of sucrose diet on apolipoprotein E biosynthesis in rat liver. Increase in apolipoprotein E gene transcription. J Biol Chem 264: 1190–1194, 1989

    Google Scholar 

  12. Driscoll DM, Mazzone T, Matsushima T, Getz G: Apoprotein E biosynthesis in the cholesterol-fed guinea pig. Arteriosclerosis 10: 31–39, 1990

    Google Scholar 

  13. Staels B, van Tol A, Chan L, Verhoeven G, Auwerx, J: Variable effects of different corticosteroids on plasma lipids, apolipoproteins, and hepatic apolipoprotein mRNA levels in rat. Arteriosclerosis & Thrombosis 11: 760–769, 1991

    Google Scholar 

  14. de Silva HV, Mas-Oliva J, Taylor JM, Mahley RW: Identification of apolipoprotein B-100 low density lipoproteins, apolipoprotein B-48 remnants, and apolipoprotein E — rich high density lipoproteins in the mouse. J Lipid Res 35: 1297–1310, 1994

    Google Scholar 

  15. Spady DK, Stange EF, Bihartz LE, Dietschy JM: Bile acids regulate hepatic low density lipoprotein receptor activity in the hamster by altering cholesterol flux across the liver. Proc Nail Acad Sci USA 83: 1916–1920, 1986

    Google Scholar 

  16. Srivastava RAK, Jiao S, Tang J, Pfleger B, Kitchens RT, Schonfeld G: In vivo regulation of low density lipoprotein receptor and apolipoprotein B gene expression by dietary fat and cholesterol in inbred strains of mice. Biochim Biophys Acta 1086: 29–43, 1991

    Google Scholar 

  17. Srivastava RAK, Baumann D, Schonfeld G: In vivo regulation of low density lipoprotein receptors by estrogen differs at the posttranscriptional level in rat and mouse. Eur J Biochem 216: 527–538, 1993

    Google Scholar 

  18. Lin RC: Quantification of apolipoproteins in rat serum and in cultured rat hepatocytes by enzyme-linked immunosorbent assay. Anal Biochem 154: 316–326, 1986

    Google Scholar 

  19. Ishikawa TT, MacGee J, Morrison A, Gluek CJ: Quantitative analysis of cholesterol in 5 to 20 μl of plasma. J Lipid Res 15: 286–291, 1974

    Google Scholar 

  20. Srivastava RAK, Srivastava N, Schonfeld G: Expression of low density lipoprotein receptor, apolipoprotein A-I, apolipoprotein A-II and apolipoprotein A-IV in various rat organs utilizing an efficient and rapid method for RNA isolation. Biochem Int 27: 85–95, 1992

    Google Scholar 

  21. Srivastava RAK, Schonfeld G: Using riboprobes for Northern blotting analysis. Bio/Techniques 11: 584–587, 1991

    Google Scholar 

  22. Srivastava RAK, Pfleger B, Schonfeld G: Expression of low density lipoprotein receptor, apolipoprotein B, apolipoprotein A-I and apolipoprotein A-IV messenger RNA in various mouse organs as determined by a novel RNA-excess solution hybridization assay. Biochim Biophys Acta 1090: 95–101, 1991

    Google Scholar 

  23. Srivastava RAK, Schonfeld G: Quantification of absolute amounts of cellular messenger RNA by RNA-excess solution hybridization. In: A.J. Harwood (ed.) Methods in Molecular Biology. Protocols for Gene Analysis, Vol 31. Humana Press, New Jersey, 1994, pp 281–288

    Google Scholar 

  24. Mierendorf RC, Pfeffer DC: Direct sequencing of denatured plasmid DNA. Meth Enzymol 152: 556–562, 1987

    Google Scholar 

  25. Taylor JM, Schimke RT: Synthesis of rat liver albumin in rabbit reticulocyte cell-free protein-synthesizing system. J Biol Chem 248: 7661–7668, 1973

    Google Scholar 

  26. Del Pozo R, Nervi F, Covarrubias C, Ronco B: Reversal of progesterone-induced biliary cholesterol output by dietary cholesterol and ethinylestradiol. Biochim Biophys Acta 753: 164–172, 1983

    Google Scholar 

  27. Spady DK, Turley SD, Dietschy JM: Rates of LDL uptake and cholesterol synthesis are regulated independently in the liver. J Lipid Res 26: 465–472, 1985

    Google Scholar 

  28. Turley SD, Dietschy JM: Cholesterol metabolism and excretion. In: I. Arias, H. Papen, H. Schachter, D.A. Sharfritz(eds) The Liver: Biology and Pathology. Raven Press, New York, 1982, pp 467–492

    Google Scholar 

  29. Salter AM, Hayashi R, Al-Seeni M, Brown NF, Bruce J, Sorenson O, Atkinson EA, Middleton B, Bleackley RC, Brindley DN: Effect of hypothyroidism and high fat feeding on mRNA concentrations for the low density lipoprotein receptor and on acyl-coA-cholesterol activities in rat liver. Biochem J 276: 825–832, 1991

    Google Scholar 

  30. Smit MJ, Temmerman AM, Havinga R, Kuipers F, Vonk RJ: Shortand long-term effects of biliary damage on hepatic cholesterol metabolism in the rat. Biochem J 269: 781–788, 1990

    Google Scholar 

  31. Raicht RF, Cohen BI, Shefer S, Mosbach EH: Sterol balance studies in the rat. Effects of dietary cholesterol and β-sitosterol on sterol balance and rate limiting enzymes of sterol metabolism. Biochim Biophys Acta 388: 374–384, 1975

    Google Scholar 

  32. Smit MJ, Kuipers F, Vonk RJ, Temmerman AM, Jackle S, Windler EET: Effects of dietary cholesterol on bile formation and hepatic processing of chylomicron remnant cholesterol in the rat. Hepatology 17: 445–454, 1993

    Google Scholar 

  33. Roach PD, Balasubramanian S, Hirata F, Abbey M, Szanto A, Simons LA, Nestel PJ: The low density lipoprotein receptor and cholesterol synthesis are affected differently by dietary cholesterol in the rat. Biochim Biophys Acta 1170: 165–172, 1993

    Google Scholar 

  34. Dawson PA, Hofmann SL, Westhuyzen DRV, Sudhoff TC, Brown MS, Goldstein JL: Sterol dependent repression of low density lipoprotein receptor promoter mediated by 16-base pair sequence adjacent to binding site for transcription factor Sp1. J Biol Chem 263: 3372–3379, 1988

    Google Scholar 

  35. Sudhof TC, Russell DW, Brown MS, Goldstein JL: 42 by element from LDL receptor gene confers end-product repression by sterols when inserted into viral TK promoter. Cell 48: 1061–1069, 1987

    Google Scholar 

  36. Sakai J, Hoshino A, Takahashi S, Miura Y, Ishii H, Suzuki H, Kawarabayashi Y, Yamamoto T: Structure, chromosome location, and expression of the human very low density lipoprotein receptor gene. J Biol Chem 269: 2173–2182, 1994

    Google Scholar 

  37. Srivastava RAK, Tang J, Baumann D, Schonfeld G: Hormonal and nutritional stimuli modulate apolipoprotein B mRNA editing in mice. Biochem Biophys Res Commun 188, 135–141, 1992

    Google Scholar 

  38. Fungwe TV, Cagen L, Wilcox HG, Heimberg M: Regulation of hepatic secretion of very low density lipoproteins by dietary cholesterol. J Lipid Res 33: 179–191, 1992

    Google Scholar 

  39. Hennessy LK, Osada J, Ordovas JM, Nicolosi RJ, Stucchi AP, Brousseau ME, Schaefer EJ: Effects of dietary fats and cholesterol on liver lipid content and hepatic apolipoprotein A–I, B and E and LDL receptor mRNA in cebus monkey. J Lipid Res 33: 351–360, 1992

    Google Scholar 

  40. Zechner R, Moser R, Newman TC, Fried SK, Breslow JL: Apolipoprotein E gene expression in mouse 3T3-L1 adipocytes and human adipose tissue and its regulation by differentiation and lipid content. J Biol Chem 266: 10583–10588, 1991

    Google Scholar 

  41. Paik Y, Chang DJ, Reardon CA, Walker MD, Taxman E, Taylor, JM: Identification and characterization of transcriptional regulatory regions associated with expression of the human apolipoprotein E gene. J Biol Chem 263: 13340–13349, 1988

    Google Scholar 

  42. Smith JD, Melian A, Leff T, Breslow JL: Expression of the human apolipoprotein E gene is regulated by multiple positive and negative elements. J Biol Chem 263: 8300–8308, 1988

    Google Scholar 

  43. Dixon J, Furukawa S, Ginsberg H: Oleate stimulates secretion of apolipoprotein B-containing lipoproteins from HepG2 cells by inhibiting early intracellular degradation of apolipoprotein B. J Biol Chem 266: 5080–5086, 1991

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Atherosclerosis, Nutrition and Lipid Research, Department of Internal Medicine, Washington University School of Medicine, Box 8046, 63110, Saint Louis, MO, USA

    Rai Ajit K. Srivastava

Authors
  1. Rai Ajit K. Srivastava
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Srivastava, R.A.K. Regulation of the apolipoprotein E by dietary lipids occurs by transcriptional and post-transcriptional mechanisms. Mol Cell Biochem 155, 153–162 (1996). https://doi.org/10.1007/BF00229312

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00229312

Key words

Search

Navigation