In Vitro Effects of Dexamethasone on Human Serum Paraoxonase-I PON1 Activity

Year 2009, Volume: 37 Issue: 3, 197 - 205, 01.08.2009

Şükrü Beydemir Deniz Ekinci Orhan Ateş

Abstract

Dexamethasone has successfully been used for treatment of some inflammatory diseases. Here, we evaluated the in vitro effects of dexamethasone on the activity of human serum paraoxonase hPON1 . PON1 enzyme was purified using simple chromatographic methods: DEAE-Sephadex anion exchanger and Sephadex G 200 gel filtration chromatography from human serum. hPON1 was purified approx. 225-fold with a final specific activity of 4867.3 U x mg-1 proteins and with a purity up to 39.8%. Dexamethasone dose-dependently decreased in vitro hPON1 activity. IC50 value for dexamethasone was determined as 1.106. Inhibition constant Ki was estimated as 3.284 ± 0.394 mM from Lineweaver-Burk graph. Inhibition type of dexamethasone was determined as noncompetitive. Our results show that dexamethasone inhibits hPON1 activity under in vitro conditions.

Keywords

Paraoxonase, Dexamethasone, Human serum, Inhibition, Enzyme purification

References

• 1. Beydemir, Ş., Kulaçoğlu, D.N., Çiftçi, M., Küfrevioğlu, Ö.İ., The in vitro effects of dexamethasone on sheep lens glucose-6-phosphate dehydrogenase, Turk. J. Chem., 27, 601, 2003.
• 2. Ekinci, D., Beydemir, Ş., Alım, Z., Some drugs inhibit in vitro hydratase and esterase activities of human carbonic anhydrase-I and II, Pharmacol. Rep., 59, 580, 2007.
• 3. Aharoni, A., Gaidukov, L., Khersonsky, O., Gould, S.M., Roodveldt, C., The ‘evolvability’ of promiscuous protein functions, Nat. Genet., 37, 73, 2005.
• 4. Alici, H.A., Ekinci, D, Beydemir, Ş., Intravenous anesthetics inhibit human paraoxonase-I (PON1) activity in vitro and in vivo, Clin. Biochem., 41(16-17) 1384, 2008.
• 5. Khersonsky, O., Tawfik, D.S., Structure-reactivity studies of serum paraoxonase PON1 suggest that its native activity is lactonase, Biochemistry-US., 44, 6371, 2005.
• 6. Rochua, D., Chabriere, E., Massona, P., Human paraoxonase: A promising approach for pre-treatment and therapy of organophosphorus poisoning, Toxicology, 233, 47, 2007.
• 7. Akgur, S.A., Ozturk, P., Solak, I., Moral, A.R., Human serum paraoxonase (PON1) activity in acute organophosphorus insecticide poisoning, Forensic Sci. Int., 133, 136, 2003.
• 8. Billecke, S., Draganov, D., Councell, R., Stetson, P., Watson, C., Hsu, C., La Du, B.N., Human serum paraoxonase (PON1) isozymes Q and R hydrolyze lactones and cyclic carbonate esters, Drug Metab. Dispos., 28, 1335, 2000.
• 9. Davies, H.G., Richter, R.J., Keifer, M., et al., The effect of the human serum paraoxonase polymorphism is reversed with diazoxon, soman and sarin, Nat. Genet., 14, 334, 1996.
• 10. La Du, B.N., Human serum paraoxonase/arylesterase. In Pharmacogenetics of Drug Metabolism (Kalow, W., ed.), Pergamon Elmford, NY, pp: 51–91, 1992.
• 11. Abbott, C.A., Mackness, M.I., Kumar, S., Boulton, A.J., Durrington, P.N. Serum paraoxonase activity, concentration and phenotype distribution in diabetes mellitus and its relationship to serum lipids and lipoproteins, Arterioscl. Throm. Vas., 15, 1812, 1995.
• 12. Baskol, G., Demir, H., Baskol, M., Kilic, E., Ates, F., Kocer, D., Muhtaroglu, S. Assessment of Paraoxonase 1 activity and malondialdehyde levels in patients with rheumatoid arthritis, Clin. Biochem., 38, 951, 2005.
• 13. Baskol, G., Karakucuk, S., Oner, A.O., Baskol, M., Kocer, D., Mirza, E., Saraymen, R., Ustdal, M., Serum paraoxonase 1 activity and lipid peroxidation levels in patients with age-related macular degeneration, Ophthalmologica, 220, 12, 2006.
• 14. Dantoine, T.F., Debord, J., Charmes, J.P., Merle, L., Marquet, P., Lachatre, G., Leroux-Robert, C., Decrease of serum paraoxonase activity in chronic renal failure, J. Am. Soc. Nephrol., 9, 2082, 1998.
• 15. Raiszadeh, F., Solati, M., Etemadi, A., Rahmani, M., Arabi, M., Serum paraoxonase activity before and after treatment of thyrotoxicosis, Clin. Endocrinol., 60, 75, 2004.
• 16. Ames, B.N., Shigenaga, M.K., Hagen, T.M., Oxidants, antioxidant and the degenerative diseases of aging, Proc. Natl. Acad. Sci. USA, 90, 7915, 1993.
• 17. Harman, D., Aging: phenomena and theories, Ann. N.Y. Acad. Sci. USA, 854, 1, 1998.
• 18. Watson, A.D., Berliner, J.A., Hama, S.Y., La Du, B.N., Faull, K.F., Fogelman, A.M., Navab, M., Protective effect of high density lipoprotein associated paraoxonase. Inhibition of the biological activity of minimally oxidized low density lipoprotein, J. Clin. Invest., 96, 2882, 1995.
• 19. Draganov, D.I., La Du, B., Pharmacogenetics of paraoxonases: a brief review, Naunyn. Schmiedebergs, Arch. Pharmacol., 369, 78, 2004.
• 20. Leviev, I., James, R., Simvastatin increases plasma levels of the antioxidant enzyme paraoxonase by PON1 gene activation, Atherosclerosis, 151, 41, 2000.
• 21. Malin, R., Laaksonen, R., Knuuti, J., Janatuinen, T., Vesalainen, R., Nuutila P., Lehtimäki, T., Paraoxonase genotype modifies the effect of pravastatin on highdensity lipoprotein cholesterol, Pharmacogenetics, 11, 625, 2001.
• 22. Tomás, M., Senti, M., Garcia-Faria, F., Vila, J., Torrents, A., Covas, M., Marrugat, J., Effect of simvastatin therapy on paraoxonase activity and related lipoproteins in familial hypercholesterolemic patients, Arterioscl. Throm. Vas., 20, 2113, 2000.
• 23. Beydemir, Ş., Gülçin, İ., Effects of melatonin on carbonic anhydrase from human erythrocyte in vitro and from rat erythrocyte in vivo, J. Enzym. Inhib. Med. Ch., 19, 193, 2004.
• 24. Lu, H.L., Chiang, C.H. Combined therapy of pentastarch, dexamethasone, and dibutyryl-cAMP or b2-agonist attenuates ischaemia/reperfusion injury of rat lung, Injury Int. J. Care. Injured, 39(9) 1062, 2008.
• 25. Petrellaa, A., Ercolinoa, S.F., Festaa, M., Gentilellaa, A., Toscoa, A., Conzenb, S.D., Parentea, L. Dexamethasone inhibits TRAIL-induced apoptosis of thyroid cancer cells via Bcl-xL induction, Eur. J. Cancer, 42(18) 3287, 2006.
• 26. Renault, F., Chabrière, E., Andrieu, J.P., Dublet, B., Masson, P., Rochu, D., Tandem purification of two HDLassociated partner proteins in human plasma, paraoxonase (PON1) and phosphate binding protein (HPBP) using hydroxyapatite chromatography, J. Chromatogr. B, 836, 15, 2006.
• 27. Sinan, S., Kockar, F., Arslan, O., Novel purification strategy for human PON1 and inhibition of the activity by cephalosporin and aminoglikozide derived antibiotics, Biochimie, 88, 565, 2006.
• 28. Yılmaz, H., Çiftçi, M., Beydemir, Ş., Bakan, E., Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes and investigation of some kinetic properties, Prep. Biochem. Biotech., 32, 287, 2002.
• 29. Bradford, M.M., A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 72, 248, 1976.
• 30. Laemmli, D.K., Cleavage of structural proteins during assembly of the head of bacteriophage T4, Nature, 227, 680, 1970.
• 31. Lineweaver, H., Burk, D., The determination of enzyme dissocation constants, J. Am. Chem. Soc., 57, 685, 1934.
• 32. Pla, A., Rodrigo, L., Hernandez, A.F., Gil, F., Lopez, O., Effect of metal ions and calcium on purified PON1 and PON3 from rat liver, Chem. Biol. Interact., 167, 63, 2007.
• 33. Abdollahi, M., Jalali, N., Jafari, A.A., Nikfar, S., A new approach to the efficacy of oximes in the management of acute organophosphate poisoning, Iran J. Med. Sci., 20, 105, 1995.
• 34. Shadnia, S., Azizi, E., Hosseini, R., Khoei, S., Fouladdel, S., Pajoumand, A., Jalali, N., Abdollahi, M., Evaluation of oxidative stres and genotoxicity in organophosphorus insecticide formulators, Hum. Exp. Toxicol., 9, 439, 2005.
• 35. Jaouad, L., Milochevitch, C., Khalil, A., PON1 paraoxonase activity is reduced during HDL oxidation and is an indicatr of HDL antioxidant capacity, Free Radic. Res., 37, 77, 2003.
• 36. Sorenson, R.C., Primo-Parmo, S.L., Kuo, C.L., Adkins, S., Lockridge, O., La Du, B.N., Reconsideration of the catalytic center and mechanism of mammalian paraoxonase/arylesterase, P. Natl. Acad. Sci. USA., 92, 7187, 1995.
• 37. Quéméneur, T., Martin-Nizard, F., Kandoussi, A., Kyndt, X., Vanhille, P., Hachulla, E., Hatron, P.Y., Fruchart, J.C., Duriez, P., Lambert, M., PON1, A new biomarker of cardiovascular disease, is low in patients with systemic vasculitis, Semin. Arthritis. Rheum., 37(3) 149, 2007.
• 38. Gülçin, I., Beydemir, Ş., Büyükokuroğlu, M.E., In vitro and in vivo effects of dantrolene on carbonic anhydrase enzyme activities, Biol. Pharm. Bull., 27, 613, 2004.
• 39. Furlong, C.E., Costa, L.G., Hasett, C., Richter, R.J., Sundstrom, J.A., Adler, D.A., Disteche, C.M., Omiecinski, C.J., Crabb, J.W., Humbert, R., Human and rabbit paraoxonases: purification, cloning, sequencing, mapping and role of polymorphism in organophosphate detoxification, Chem. Biol. Interact., 87, 35, 1993.