Skip to main content
SpringerLink
Log in

In Vitro Simultaneous Transfer of Lipids to HDL in Coronary Artery Disease and in Statin Treatment

  • Methods
  • Published:
Lipids

Abstract

The exchange of lipids with cells and other lipoproteins is a crucial process in HDL metabolism and for HDL antiatherogenic function. Here, we tested a practical method to quantify the simultaneous transfer to HDL of phospholipids, free-cholesterol, esterified cholesterol and triacylglycerols and to verify the lipid transfer in patients with coronary artery disease (CAD) or undergoing statin treatment. Twenty-eight control subjects without CAD, 27 with CAD and 25 CAD patients under simvastatin treatment were studied. Plasma samples were incubated with a donor nanoemulsion prepared by ultrasonication of the constituent lipids and labeled with radioactive lipids; % lipids transferred to HDL were quantified in the HDL-containing supernatant after chemical precipitation of non-HDL fractions and the nanoemulsion. The assay was precise and reproducible. Increase of temperature (4–37 °C), of incubation period (5 min to 2 h), of HDL-cholesterol concentration (33–244 mg/dL) and of mass of nanoemulsion lipids (0.075–0.3 mg/μL) resulted in increased lipid transfer from the nanoemulsion to HDL. In contrast, increasing pH (6.5–8.5) and albumin concentration (3.5–7.0 g/dL) did not affect lipid transfer. There was no difference between CAD and control non-CAD with regard to the lipid transfer, but statin treatment reduced the transfer to HDL of all four lipids. The test herein described is a valid and practical tool for exploring an important aspect of HDL metabolism.

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

Fig. 1

Similar content being viewed by others

Abbreviations

CETP:

Cholesteryl ester transfer protein

PLTP:

Phospholipid transfer protein

Apo:

Apolipoprotein

CAD:

Coronary artery disease

BMI:

Body mass index

References

  1. Tall AR (1993) Plasma cholesteryl ester transfer protein. J Lipid Res 34:1255–1274

    CAS  PubMed  Google Scholar 

  2. Tollefson JH, Ravnik S, Albers JJ (1988) Isolation and characterization of a phospholipid transfer protein (LTP-II) from human plasma. J Lipid Res 29:1593–1602

    CAS  PubMed  Google Scholar 

  3. Ferretti G, Bacchetti T, Nègre-Salvayre A, Salvayre R, Dousset N, Curatola G (2006) Structural modifications of HDL and functional consequences. Atherosclerosis 184:1–7

    Article  CAS  PubMed  Google Scholar 

  4. Norata GD, Pirillo A, Catapano AL (2006) Modified HDL: biological and physiopathological consequences. Nutr Metab Cardiovasc Dis 16:371–386

    Article  CAS  PubMed  Google Scholar 

  5. Lamarche B, Rashid S, Lewis GF (1999) HDL metabolism in hypertriglyceridemic states: an overview. Clin Chim Acta 286:145–149

    Article  CAS  PubMed  Google Scholar 

  6. Rashid S, Watanabe T, Sakaue T, Lewis GF (2003) Mechanisms of HDL lowering in insulin resistant, hypertriglyceridemic states: the combined effect of HDL triglyceride enrichment and elevated hepatic lipase activity. Clin Biochem 36:421–429

    Article  CAS  PubMed  Google Scholar 

  7. Assmann G, Gotto AM Jr (2004) HDL cholesterol and protective factors in atherosclerosis. Circulation 109(23 Suppl 1):III8–14

    PubMed  Google Scholar 

  8. Kontush A, Chapman MJ (2006) Antiatherogenic small, dense HDL–guardian angel of the arterial wall? Nat Clin Pract Cardiovasc Med 3:144–153

    Article  CAS  PubMed  Google Scholar 

  9. Friedewald WT, Levy RI, Fredrickison DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without the use of the preparative ultracentrifuge. Clin Chem 18:499–502

    CAS  PubMed  Google Scholar 

  10. Lima E, Maranhão RC (2004) Rapid, simple laser-light scattering method for HDL particle size in whole plasma. Clin Chem 50:1086–1091

    Article  CAS  PubMed  Google Scholar 

  11. Ginsburg GS, Small DM, Atkinson D (1982) Microemulsions of phospholipids and cholesterol ethers protein-free models of low-density lipoprotein. J Biol Chem 257:8216–8221

    CAS  PubMed  Google Scholar 

  12. Maranhão RC, Cesar TB, Pedroso-Mariani SR (1993) Metabolic behavior in rats of a nonprotein microemulsion resembling LDL. Lipids 28:691–696

    Article  PubMed  Google Scholar 

  13. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358

    Article  CAS  PubMed  Google Scholar 

  14. Guidance for Industry (2001) Bioanalytical method validation. US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER) and Center for Veterinary Medicine (CVM)

  15. Cutri BA, Hime NJ, Nicholls SJ (2006) High-density lipoproteins: an emerging target in the prevention of cardiovascular disease. Cell Res 16:799–804

    Article  CAS  PubMed  Google Scholar 

  16. Barter PJ, Rye KA (2006) Relationship between the concentration and antiatherogenic activity of high-density lipoproteins. Curr Opin Lipid 17:399–403

    Article  CAS  Google Scholar 

  17. Lewis GF (2006) Determinants of plasma HDL concentrations and reverse cholesterol transport. Curr Opin Cardiol 21:345–352

    Article  PubMed  Google Scholar 

  18. Futterman LG, Lemberg L (2005) Apo a-I Milano. Am J Crit Care 14:244–247

    PubMed  Google Scholar 

  19. Dullens SP, Plat J, Mensink RP (2007) Increasing apoA-I production as a target for CHD risk reduction. Nutr Metab Cardiovasc Dis 17:616–628

    Article  CAS  PubMed  Google Scholar 

  20. Tall AR, Yvan-Charvet L, Wang N (2007) The failure of torcetrapib: was it the molecule or the mechanism? Arterioscler Thromb Vasc Biol 27:257–260

    Article  CAS  PubMed  Google Scholar 

  21. Cheung MC, Wolfbauer G, Brown BG, Albers JJ (1999) Relationship between plasma phospholipid transfer protein activity and HDL subclasses among patients with low HDL and cardiovascular disease. Atherosclerosis 142:201–205

    Article  CAS  PubMed  Google Scholar 

  22. Tall AR, Forester LR, Bongiovanni GL (1983) Facilitation of phosphatidylcholine transfer into high density lipoproteins by an apolipoprotein in the density 1.20–1.26 g/mL fraction of plasma. J Lipid Res 24:277–289

    CAS  PubMed  Google Scholar 

  23. Lassel TS, Guerin M, Auboiron S, Guy-Grand B, Chapman MJ (1999) Evidence for a cholesteryl ester donor activity of LDL particles during alimentary lipemia in normolipidemic subjects. Atherosclerosis 147:41–48

    Article  CAS  PubMed  Google Scholar 

  24. Clark RW, Ruggeri RB, Cunningham D, Bamberger MJ (2006) Description of the torcetrapib series of cholesteryl ester transfer protein inhibitors, including mechanism of action. J Lipid Res 47:537–552

    Article  CAS  PubMed  Google Scholar 

  25. Terpstra AH, Nicolosi RJ, Herbert PN (1989) In vitro incorporation of radiolabeled cholesteryl esters into high and low density lipoproteins. J Lipid Res 30:1663–1671

    CAS  PubMed  Google Scholar 

  26. Sammett D, Tall AR (1985) Mechanisms of enhancement of cholesteryl ester transfer protein activity by lipolysis. J Biol Chem 260:6687–6697

    CAS  PubMed  Google Scholar 

  27. Desrumaux C, Athias A, Masson D, Gambert P, Lallemant C, Lagrost L (1998) Influence of the electrostatic charge of lipoprotein particles on the activity of the human plasma phospholipid transfer protein. J Lipid Res 39:131–142

    CAS  PubMed  Google Scholar 

  28. Soares TA, Ferreira R (2004) Aplicação da equação de Poisson–Boltzmann ao cálculo de propriedades dependentes do pH em proteínas. Quim Nova 27:640–647

    CAS  Google Scholar 

  29. Sparks DL, Pritchard PH (1989) Transfer of cholesteryl ester into high density lipoprotein by cholesteryl ester transfer protein: effect of HDL lipid and apoprotein content. J Lipid Res 30:1491–1498

    CAS  PubMed  Google Scholar 

  30. Lagrost L, Florentin E, Guyard-Dangremont V, Athias A, Gandjini H, Lallemant C, Gambert P (1995) Evidence for nonesterified fatty acids as modulators of neutral lipid transfers in normolipidemic human plasma. Arterioscler Thromb Vasc Biol 15:1388–1396

    CAS  PubMed  Google Scholar 

  31. Estronca LM, Moreno MJ, Laranjinha JA, Almeida LM, Vaz WL (2005) Kinetics and thermodynamics of lipid amphiphile exchange between lipoproteins and albumin in serum. Biophys J 88:557–565

    Article  CAS  PubMed  Google Scholar 

  32. Blankenberg S, Rupprecht HJ, Bickel C, Jiang XC, Poirier O, Lackner KJ, Meyer J, Cambien F, Tiret L (2003) Common genetic variation of the cholesteryl ester transfer protein gene strongly predicts future cardiovascular death in patients with coronary artery disease. J Am Coll Cardiol 41:1983–1989

    Article  CAS  PubMed  Google Scholar 

  33. Goto A, Sasai K, Suzuki S, Fukutomi T, Ito S, Matsushita T, Okamoto M, Suzuki T, Itoh M, Okumura-Noji K, Yokoyama S (2001) Cholesteryl ester transfer protein and atherosclerosis in Japanese subjects: a study based on coronary angiography. Atherosclerosis 159:153–163

    Article  CAS  PubMed  Google Scholar 

  34. Boekholdt SM, Kuivenhoven JA, Wareham NJ, Peters RJG, Jukema JW, Luben R, Bingham SA, Day NE, Kastelein JJ, Khaw KT (2004) Plasma levels of cholesteryl ester transfer protein and the risk of future coronary artery disease in apparently healthy men and women. Circulation 110:1418–1423

    Article  CAS  PubMed  Google Scholar 

  35. Schlitt A, Bickel C, Thumma P, Blankenberg S, Rupprecht HJ, Meyer J, Jiang XC (2003) High plasma phospholipid transfer protein levels as a risk factor for coronary artery disease. Arterioscler Thromb Vasc Biol 23:1857–1862

    Article  CAS  PubMed  Google Scholar 

  36. Schgoer W, Mueller T, Jauhiainen M, Wehinger A, Gander R, Tancevski I, Salzmann K, Eller P, Ritsch A, Haltmayer M, Ehnholm C, Patsch JR, Foeger B (2008) Low phospholipid transfer protein (PLTP) is a risk factor for peripheral atherosclerosis. Atherosclerosis 196:219–226

    Article  CAS  PubMed  Google Scholar 

  37. Ahnadi CE, Berthezène F, Ponsin G (1993) Simvastatin-induced decrease in the transfer of cholesterol esters from high density lipoproteins to very low and low density lipoproteins in normolipidemic subjects. Atherosclerosis 99:219–228

    Article  CAS  PubMed  Google Scholar 

  38. McPherson R (1999) Comparative effects of simvastatin and cholestyramine on plasma lipoproteins and CETP in humans. Can J Clin Pharmacol 6:85–90

    CAS  PubMed  Google Scholar 

  39. de Vries R, Kerstens MN, Sluiter WJ, Groen AK, van Tol A, Dullaart RP (2005) Cellular cholesterol efflux to plasma from moderately hypercholesterolaemic type 1 diabetic patients is enhanced, and is unaffected by simvastatin treatment. Diabetologia 48:1105–1113

    Article  CAS  PubMed  Google Scholar 

  40. Sviridov D, Nestel P, Watts G (2007) Statins and metabolism of high density lipoprotein. Cardiovasc Hematol Agents Med Chem 5:215–221

    Article  CAS  PubMed  Google Scholar 

  41. Guerin M, Lassel TS, Le Goff W, Farnier M, Chapman MJ (2000) Action of atorvastatin in combined hyperlipidemia: preferential reduction of cholesteryl ester transfer from HDL to VLDL 1 particles. Arterioscler Thromb Vasc Biol 20(1):189–197

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Fundação do Amparo à Pesquisa do Estado de São Paulo (FAPESP), São Paulo, Brazil (Grant 0408048-3). Dr. Maranhão has a Research Award from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brasilia, Brazil. The authors are grateful to Débora Deus for help with the experiments and to Dr. Fatima R. Freitas for her help in revising the manuscript.

Author information

Authors and Affiliations

  1. Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil

    Ana C. Lo Prete, Carolina H. Azevedo & Raul Cavalcante Maranhão

  2. Laboratório de Metabolismo de Lípides, Instituto do Coração (INCOR) do Hospital das Clínicas FMUSP, Av. Dr. Enéas de Carvalho Aguiar, 44, 1º subsolo, São Paulo, SP, 05403-000, Brazil

    Clederson H. Dina, Camila G. Puk, Neuza H. M. Lopes, Whady A. Hueb & Raul Cavalcante Maranhão

Authors
  1. Ana C. Lo Prete
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Clederson H. Dina
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Carolina H. Azevedo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Camila G. Puk
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Neuza H. M. Lopes
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Whady A. Hueb
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Raul Cavalcante Maranhão
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Raul Cavalcante Maranhão.

About this article

Cite this article

Lo Prete, A.C., Dina, C.H., Azevedo, C.H. et al. In Vitro Simultaneous Transfer of Lipids to HDL in Coronary Artery Disease and in Statin Treatment. Lipids 44, 917–924 (2009). https://doi.org/10.1007/s11745-009-3342-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11745-009-3342-2

Keywords

Search

Navigation