Caffeine Increases Apolipoprotein A-1 and Paraoxonase-1 but not Paraoxonase-3 Protein Levels in Human-Derived Liver (HepG2) Cells

Abstract

Background: Apolipoprotein A-1, paraoxonase-1 and paraoxonase-3 are antioxidant and anti-atherosclerotic structural high-density lipoprotein proteins that are mainly synthesized by the liver. No study has ever been performed to specifically examine the effects of caffeine on paraoxonase enzymes and on liver apolipoprotein A-1 protein levels. Aims: To investigate the dose-dependent effects of caffeine on liver apolipoprotein A-1, paraoxonase-1 and paraoxonase-3 protein levels. Study Design: In vitro experimental study.Methods: HepG2 cells were incubated with 0 (control), 10, 50 and 200 μM of caffeine for 24 hours. Cell viability was evaluated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. Apolipoprotein A-1, paraoxonase-1 and paraoxonase-3 protein levels were measured by western blotting.Results: We observed a significant increase on apolipoprotein A-1 and paraoxonase-1 protein levels in the cells incubated with 50 µM of caffeine and a significant increase on paraoxonase-1 protein level in the cells incubated with 200 µM of caffeine.Conclusion: Our study showed that caffeine does not change paraoxonase-3 protein level, but the higher doses used in our study do cause an increase in both apolipoprotein A-1 and paraoxonase-1 protein levels in liver cells.

Keywords

• Caffeine, apolipoprotein A-1, paraoxonase-1, paraoxonase-3, HepG2 cells

References

1. 1. Singh U, Jialal I. Oxidative stres and atherosclerosis. Pathophysiology 2006;13:129-42.
2. 2. Aviram M. Macrophage foam cell formation during early atherogenesis is determined by the balance between pro-oxidants and anti-oxidants in arterial cells and blood lipoproteins. Antioxid Redox Signal 1999;1:585- 94.
3. 3. Aviram M, Kaplan M, Rosenblat M, Fuhrman B. Dietary antioxidants and paraoxonases against LDL oxidation and atherosclerosis development. Handb Exp Pharmacol 2005;170:263-300.
4. 4. Podrez EA. Anti-oxidant properties of high-density lipoprotein and atherosclerosis. Clin Exp Pharmacol Physiol 2010;37:719-25.
5. 5. Précourt LP, Amre D, Denis MC, Lavoie JC, Delvin E, Seidman E, et al. The three-gene paraoxonase family: physiologic roles, actions and regulation. Atherosclerosis 2011;214:20-36.
6. 6. Aviram M, Rosenblat M. Paraoxonases 1, 2, and 3, oxidative stress, and macrophage foam cell formation during atherosclerosis development. Free Radic Biol Med 2004;37:1304-16.
7. 7. Draganov DI, Stetson PL, Watson CE, Billecke SS, La Du BN. Rabbit serum paraoxonase 3 (PON3) is a high density lipoprotein-associated lactonase and protects low density lipoprotein against oxidation. J Biol Chem 2000;275:33435-42.
8. 8. Reddy ST, Wadleigh DJ, Grijalva V, Ng C, Hama S, Gangopadhyay A, et al. Human paraoxonase-3 is an HDL-associated enzyme with biological activity similar to paraoxonase-1 protein but is not regulated by oxidized lipids. Arterioscler Thromb Vasc Biol 2001;21:542-7.

9. 9. Clark I, Landolt HP. Coffee, caffeine, and sleep: A systematic review of epidemiological studies and randomized controlled trials. Sleep Med Rev 2016;31:70-8.
10. 10. Winston AP, Hardwick E, Jaberi N. Neuropsychiatric effects of caffeine. Adv Psychiatr Treat 2005;11:432-9.
11. 11. Lane JD, Pieper CF, Barefoot JC, Williams RB Jr, Siegler IC. Caffeine and cholesterol: interactions with hostility. Psychosom Med 1994;56:260-6.
12. 12. Papamichael CM, Aznaouridis KA, Karatzis EN, Karatzi KN, Stamatelopoulos KS, Vamvakou G, et al. Effect of coffee on endothelial function in healthy subjects: the role of caffeine. Clin Sci (Lond) 2005;109:55-60.
13. 13. Reis JP, Loria CM, Steffen LM, Zhou X, van Horn L, Siscovick DS, et al. Coffee, decaffeinated coffee, caffeine, and tea consumption in young adulthood and atherosclerosis later in life: the CARDIA study. Arterioscler Thromb Vasc Biol 2010;30:2059-66.
14. 14. Sedor FA, Schneider KA, Heyden S. Effect of coffee on cholesterol and apolipoproteins, corroborated by caffeine levels. Am J Prev Med 1991;7:391-6.
15. 15. Carson CA, Cauley JA, Caggiula AW. Relation of caffeine intake to blood lipids in elderly women. Am J Epidemiol 1993;138:94-100.
16. 16. Kempf K, Herder C, Erlund I, Kolb H, Martin S, Carstensen M, et al. Effects of coffee consumption on subclinical inflammation and other risk factors for type 2 diabetes: a clinical trial. Am J Clin Nutr 2010;91:950-7.
17. 17. Bouma ME, Rogier E, Verthier N, Labarre C, Feldmann G. Further cellular investigation of the human hepatoblastoma-derived cell line HepG2: morphology and immunocytochemical studies of hepatic-secreted proteins. In Vitro Cell Dev Biol 1989;25:267-75.
18. 18. Jaichander P, Selvarajan K, Garelnabi M, Parthasarathy S. Induction of paraoxonase 1 and apolipoprotein A-I gene expression by aspirin. J Lipid Res 2008;49:2142-8.
19. 19. Kulkarni PB, Dorand RD. Caffeine toxicity in a neonate. Pediatrics 1979;64:254-5.
20. 20. Fredholm BB, Bättig K, Holmén J, Nehlig A, Zvartau EE. Actions of caffeine in the brain with special reference to factors that contribute to its widespread use. Pharmacol Rev 1999;51:83-133.
21. 21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
22. 22. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
23. 23. Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012;9:671-5.
24. 24. Nehlig A, Daval JL, Debry G. Caffeine and the central nervous system: mechanisms of action, biochemical, metabolic and psychostimulant effects. Brain Res Rev 1992;17:139-70.
25. 25. Azam S, Hadi N, Khan NU, Hadi SM. Antioxidant and prooxidant properties of caffeine, theobromine and xanthine. Med Sci Monit 2003;9:325-30.
26. 26. Yukawa GS, Mune M, Otani H, Tone Y, Liang XM, Iwahashi H, et al. Effects of coffee consumption on oxidative susceptibility of low-density lipoproteins and serum lipid levels in humans. Biochemistry (Mosc) 2004;69:70-4.
27. 27. Moore RE, Navab M, Millar JS, Zimetti F, Hama S, Rothblat GH, et al. Increased atherosclerosis in mice lacking apolipoprotein A-I attributable to both impaired reverse cholesterol transport and increased inflammation. Circ Res 2005;97:763-71.
28. 28. De Geesr B, Stengel D, Landeloos M, Lox M, Le Gat L, Collen D, et al. Effect of overexpression of human apo A-I in C57BL/6 and C57BL/6 apo E-deficient mice on 2 lipoprotein-associated enzymes, platelet-activating factor acetylhydrolase and paraoxonase. Comparison of adenovirusmediated human apo A-I gene transfer and human apo A-I transgenesis. Arterioscler Thromb Vasc Biol 2000;20:68-75.
29. 29. Ikeda Y, Suehiro T, Arii K, Kumon Y, Hashimoto K. High glucose induces transactivation of the human paraoxonase 1 gene in hepatocytes. Metabolism 2008;57:1725-32.
30. 30. Ozgun E, Ozgun GS, Gokmen SS, Eskiocak S, Sut N, Akıncı M, et al. Effect of lipoic acid on serum paraoxonase-1 and paraoxonase-3 protein levels and activities in diabetic rats. Exp Clin Endocrinol Diabetes 2016.