Urine 8-Hydroxy-2’-Deoxyguanosine (8-OHdG) Levels of Dogs in Pyoderma

Abstract

In this study, it is aimed to investigate correlation between antioxidative metabolism and the significant skin disease pyoderma in dogs and to demonstrate the extent of oxidative damage with 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. Healthy (n=13), females diagnosed as pyoderma (untreated) (n=16), heathy (n=14) males and males diagnosed as pyoderma (untreated) (n=12) consisted the four study groups. In this aspect two control and two diseased groups were formed according to gender. Blood and urine samples were obtained from all the groups and bacterial swabs were collected from the diseased groups as well. Plasma malondialdehyde (MDA) besides urine 8-hydroxy-2'-deoxyguanosine levels measured with LC/MS/MS. MDA and 8OHdG levels were increased in diseased groups. Differences among groups were found to be significant (p ≤ 0.05). In conclusion in our opinion untreated pyoderma cases could result in DNA damage

Keywords

• Dog
• pyoderma
• oxidative stress
• MDA
• 8-OHdG

Piyodermalı köpeklerde idrarda 8-hidroksi-2’-deoksiguanozin (8-OHdG) düzeyleri

Abstract

Bu çalışmada, köpeklerin önemli bir deri hastalığı olan piyoderma ile antioksidatif metabolizma arasındaki ilişkinin incelenmesi ve oksidatif hasarın boyutunun 8-hidroksi-2’-deoksiguanozin (8-OHdG) üzerinden gösterilmesi amaçlanmıştır. Çalışmada sağlıklı dişi köpekler (n=13) ile piyoderma tanısı konmuş (tedavi almamış) dişi köpekler (n=16) ve sağlıklı erkek köpekler (n=14) ile piyoderma tanısı konmuş (tedavi almamış) erkek köpekler (n=12) olmak üzere cinsiyete göre iki kontrol ve iki hasta grubu oluşturulmuştur. Hasta gruptan kan, idrar ve svap, sağlıklı gruptan kan ve idrar örnekleri alınmıştır. Plazmada malondialdehit (MDA) ve idrarda LC/MS/MS ile 8-hidroksi-2’-deoksiguanozin düzeyleri ölçülmüştür. Piyoderma tanısı konmuş erkek ve dişi köpeklerde MDA ve 8-OHdG düzeylerinin yükseldiği saptanmıştır. Bu artış, istatistik olarak da anlamlı bulunmuştur (p ≤ 0.05). Sonuç olarak piyodermaların, tedavi edilmediği takdirde DNA hasarına yol açabileceği kanısına varılmıştır

Keywords

• Köpek
• piyoderma
• oksidatif stres
• MDA
• 8-OHdG

References

1. Aiello SE (1998): The Merck Veterinary Manual. Eighth Edition. U.S.A.
2. Bickers DR, Athar M (2006): Oxidative stress in the pathogenesis of skin diseases. J Invest Dermatol, 126. 2565-2575.
3. Crow B, Bishop M, Kovalcık K, Norton D, George J, Bralley A (2008): A simple and cost effective method for the quantification of 8-hydroxy-2’-deoxyguanosine from urine using liquid chromatography tandem mass spectrometry. Biomed Chromatogr, 22, 394-401.
4. Dizdaroğlu M, Gajewski E (1990): Selected-Ion Mass Spectrometry: Assays of Oxidative DNA Damage. Method Enzymol, 186, 530-544.
5. Dizdaroğlu M, Jaruga P, Rodriguez H (2001): Measurement of 8-hydroxy-2’-deoxyguanosine in DNA by high-performance liquid chromatography-mass spectrometry: comparison with measurement by gas chromatography-mass spectrometry. Nucleic Acids Res, 29,E12.
6. Fidancı UR, Turgay F, Zengin S, Kargın F, Çelik S, Taşdemir U (2001): Genotipin Ankara Keçilerinde antioksidatif Metabolizma Üzerine Etkisi. Turk J Vet Anim Sci, 25, 975-981.
7. Hattori Y, Nishigori C, Tanaka T, Uchida K, Nikaido O, Osawa T, Hiai H, Imamura S, Toyokuni S (1996): 8- hydroxy-2’-deoxyguanosine is increased in epidermal cells of hairless mice after chronic ultraviolet B exposure. J.Invest.Dermatol, 107, 733-737.
8. Helbock HJ, Beckman KB, Shigenaga MK, Walter PB, Woodall AA, Yeo HC, Ames BN (1998): DNA oxidation matters: The HPLC-electrochemical detection assay of 8- oxo-deoxyguanosine and 8-oxo-guanine. Proc Natl Acad Sci, 95, 288-293.

9. Hill PB, Moriello KA (1994): Canine Pyoderma. Javma, 204, 334-340.
10. Horvath C, Neuber A (2007): Management of canine pyoderma. Uk Vet, 12 , 1-7.
11. Jalel A, Yassine M, Hamdaoui M (2009): Oxidative stres in experimental vitiligo C57BL/6 mice. Indian J Dermatol, 54, 221-224.
12. Janero D (1998): Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radical Biol Med, 9, 515-540.
13. Jaruga P, Rodriguez H, Dizdaroğlu M (2001): Measurement of 8-hydroxy-2’-deoxyadenosine in DNA by liquid chromatography/mass spectometry. Free Radical Biol Med, 31, 336-344.
14. Jentzsh AM, Bachmann H, Furst P, Biesalski HK (1996): Improved analysis of malondialdehyde in human body fluids. Free Radical Biol Med, 20, 251-6.
15. Jo C, Ahn DU. (1998): Fluorometric analysis of 2- thiobarbituric acid reactive substances in turkey. Pout Sci March, 77, 475-80.
16. Karaca Ş, Güder H (2009): Dermatolojide Antioksidan Sistem. Turk J Dermatol, 3, 32-39.
17. Kutsal A, Alpan O, Arpacık R, (1990): İstatistik Uygulamalar. X+231. Bizim Büro Basımevi. Ankara.
18. Lin H, Jenner A, Ong C, Huang S, Whiteman M, Halliwell B (2004): A high-throughput and sensitive methodology for the quantificaition of urinary 8-hydroxy- 2’-deoxyguanosine: measurement with gas chromatography-mass spectrometry after single solid- phase exraction. Biochem J, 380, 541-548.
19. Matsufuji H, Ochi H, Shibamoto T (2006): Formation and inhibition of genotoxic malonaldehyde from DNA oxidation controlled with EDTA. Food Chem Toxicol, 44, 236-241.
20. Murtas D, Piras F, Minerba L, Ugalde J, Floris C, Maxia C, Demurtas P, Perra MT, Sirigu P (2010): Nuclear 8-hydroxy-2'-deoxyguanosine as survival biomarker in patients with cutaneous melanoma. Oncol Rep, 23, 329-35.
21. Nakai K, Yoneda K, Maeda R, Munehiro A, Fujita N, Yokoi I, Moriue J, Moriue T, Kosaka H, Kubata Y (2009): Urinary biomarker of oxidative stress in patients with psoriasis vulgaris and atopic dermatitis. J Eur Acad Dermatol Venerol, Dec; 23,1405-1408.
22. Pietta PG, Simonetti P, Gardana C, Cristoni S, Bramati L, Mauri PL (2003): LC-APCI-MS/MS analysis of urinary 8-hydroxy-2’-deoxyguanosine. J Pharmaceut Biomed, 32, 657-661.
23. Pilger A, Rudiger HW (2006): 8-hydroxy-2’- deoxyguanosine as a marker of oxidative DNA damage related to occupational and environmental exposures. Int Arch Occup Environ Health, 80, 1-15.
24. Şentürk S, Özel E, Şen A (2005): Clinical Efficacy of Rifampicin for Treatment of Canine Pyoderma. Acta Vet Brno, 74, 117-122.
25. Tüközkan N, Erdamar H, Seven I (2006): Measurement of total malondialdehyde in plasma and tissues by high- performanced liquid chromatgraphy and thiobarbituric acid assay. Fırat tıp dergisi, 11,88-92.
26. Tsuboi H, Kouda K, Takeuchi H, Takigawa M, Masamoto Y, Takeuchi M, Ochi H (1998): 8- Hydroxydeoxyguanosine in urine as an index of oxidative damage to DNA in the evaluation of atopic dermatitis. Brit J Dermatol, 138, 1033-1035.
27. Zeptometrix TBARS Assay Kit (2010): Erişim Tarihi: 13.06.2010. Erişim adresi: http://www.zeptometrix.com/0801192.pdf
28. Zhang F, Stott WT, Clark AJ, Schisler MR, Grundy JJ, Gollapudi BB, Bartels MJ (2007): Quantitation of 8- hydroxydeoxyguanosine in DNA by liquid chromatography/positive atmospheric pressure photoionization tandem mass spectometry. Rapid Commun Mass Spectrom, 21, 3949-3955. Geliş tarihi: 25.12.2011 / Kabul tarihi: 14.03.2012