ASSESSMENT OF DNA STRAND BREAKS AND TOTAL ANTIOXIDANT STATUS IN TYPE 2 DIABETIC PATIENTS WITH AND WITHOUT COMPLICATIONS- A CASE-CONTROL STUDY
Objective: To evaluate the total antioxidant status (TAS) and the extent of DNA strand breaks (damage) as a measure of oxidative stress biomarkers in Type 2 diabetic patients (with and without complications) and controls.
Materials and Methods: Blood samples were collected from 200 patients with type 2 diabetes mellitus (n=100 with complications and n=100 without complications) and 100 healthy individuals. Oxidative DNA damage was evaluated using alkaline single cell gel electrophoresis (comet assay). Total antioxidant status was assessed by Ferric Reducing Ability of Plasma (FRAP) assay.
Results: TAS was found to be significantly lower in type 2 diabetic patients (with and without complications) compared to controls (p< 0.001). Â Similarly, patients with complications of type 2 diabetes mellitus had significantly lower TAS when compared to diabetic patients (p= 0.007). DNA damage analysis showed that the extent of damage was high in patients with diabetes mellitus (with and without complications) compared to controls (p< 0.001). Fasting glucose and glycosylated haemoglobin level (HbA1c) was found to be significantly higherin diabetic patients than controls (p< 0.05). Correlation analysis showed that there is no association between age, duration, sugar level, HbA1c, TAS and DNA damage in patients with Type 2 diabetes mellitus.
Conclusion: Alterations in TAS and the extent of DNA damage was observed in patients with complications of diabetes mellitus indicate that oxidative stress is more in patients with complications when compared to patients without complications and healthy individuals. Therefore, further DNA damage and onset of complications in Type 2 diabetes mellitus could be prevented by counteracting the oxidative stress by therapeutic interventions using appropriate antioxidants.
Key words: Hyperglycemia, Oxidative stress, DNA damage, Total antioxidant status, Type 2 diabetes mellitus, Vascular complication
2. Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature 2001; 414 (6865): 813-820.
3. Brownlee M, Cerami A and Vlassara H. Advanced glycosylation end products in tissue and the biochemical basis of diabetic complications. N Engl J Med 1988; 318 (20): 1315-1321.
4. Greene DA, Lattimer SA and Sima AA. Sorbitol, phosphoinositides and sodium-potassium-ATPase in the pathogenesis of diabetic complications. N Engl J Med 1987; 316 (10): 599-606.
5. Koya D and King GL. Protein kinase C activation and the development of diabetic complications. Diabetes 1998; 47 (6): 859-866.
6. Baynes JW. Role of oxidative stress in development of complications in diabetes. Diabetes 1991; 40 (4): 405-412.
7. Pan H, Chang D, Feng L, Xu F, Kuang H and Lu M. Oxidative damage to DNA and its relationship with diabetic complications. Biomed Environ Sci 2007; 20 (2): 160.
8. Kangralkar VA, Patil SD and Bandivadekar RM. Oxidative stress and diabetes: a review. IJPA 2010; 1 (1): 38-45.
9. Pham-Huy LA, He H and Pham-Huy C. Free radicals, antioxidants in disease and health. Int J Biomed Sci 2008; 4 (2): 89-96.
10. Phillips M, Cataneo RN, Cheema T and Greenberg J. Increased breath biomarkers of oxidative stress in diabetes mellitus. Clinica Chimica Acta 2004; 344 (1): 189-194.
11. Asfandiyarova N, Kolcheva N, Ryazantsev I and Ryazantsev V. Risk factors for stroke in type 2 diabetes mellitus. Diab Vasc Dis Res 2007; 3(1): 57-60.
12. Bolajoko EB, Mossanda KS, Moropane M, Adeniyi F, Akinosun O and Fasanmade A. Antioxidant and oxidative stress status in type2 diabetes and diabetic foot ulcer. SAMJ 2008; 98(8): 614-617.
13. Kashiwagi A. Complications of diabetes mellitus and oxidative stress. Japan Med Assoc J 2001; 44(12): 521-528.
14. Matough FA, Budin SB, Hamid ZA, Alwahaibi N and Mohamed J. The role of oxidative stress and antioxidants in diabetic complications. Sultan Qaboos Univ Med J 2012; 12(1): 5.
15. Maxwell SRJ, Thomason H, Sandler D, Leguen C, Baxter MA, Thorpe GHG et al. Antioxidant status in patients with uncomplicated insulinâ€dependent and nonâ€insulinâ€dependent diabetes mellitus. Eur J Clin Invest 1997; 27(6): 484-490.
16. Pinzani P, Petruzzi E, Orlando C, Gallai R, Serio M and Pazzagli M. Serum antioxidant capacity in healthy and diabetic subjects as determined by enhanced chemiluminescence. J Biolumin Chemilumin 1998; 13(5): 321-325.
17. Nandhakumar S, Parasuraman S, Shanmugam MM, Rao KR, Chand P and Bhat BV. Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay). J Pharmacol Pharmacother 2011; 2(2): 107â€“111. http://doi.org/10.4103/0976-500X.81903
18. Benzie IF and Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant powerâ€: the FRAP assay. Anal Biochem 1996; 239(1): 70-76.
19. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H and Sasaki YF. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 2000; 35(3): 206-221.
20. Singh NP, McCoy MT, Tice RR and Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 1988; 175(1): 184-191.
21. Dosoo DK, Rana SV, Offe-Amoyaw K, Tete-Donkor D and Maddy SQ. Total antioxidant status in non-insulin-dependent diabetes mellitus patients in Ghana. West Afr J Med 2000; 20(3): 184-186.
22. Akinosun OM and Bolajoko EB. Total antioxidant status in type 2 diabetic patients: experience at University College Hospital (UCH) Ibadan, Nigeria. Niger J Clin Pract 2007; 10(2): 126-129.
23. Song F, Jia W, Yao Y, Hu Y, Lei L, Lin J, Sun X and Liu L. Oxidative stress, antioxidant status and DNA damage in patients with impaired glucose regulation and newly diagnosed Type 2 diabetes. Clin Sci 2007; 112(12): 599-606.
24. Lodovici M, Giovannelli L, Pitozzi V, Bigagli E, Bardini G and Rotella CM. Oxidative DNA damage and plasma antioxidant capacity in type 2 diabetic patients with good and poor glycaemic control. Mutat Res Fundam Mol Mech Mutagen 2008; 638(1): 98-102.
25. Rani AJ and Mythili SV. Study on total antioxidant status in relation to oxidative stress in type 2 diabetes mellitus. J Clin Diagn Res 2014; 8(3): 108-110.
26. Balakumaran TA and Seshadri S. Antioxidants and lipid peroxidation status in diabetic patients with and without complications. Arch Iranian Med 2009; 12(2): 121-127.
27. Opara EC, Abdel-Rahman E, Soliman S, Kamel WA, Souka S, Lowe JE and Abdel-Aleem S. Depletion of total antioxidant capacity in type 2 diabetes. Metabolism 1999; 48(11): 1414-1417.
28. Dordevic G, Duric S, Apostolski S, Dordevic V and Zivkovic M. Total antioxidant blood capacity in patients with type 2 diabetes mellitus and distal symmetrical polyneuropathy. Vojnosanitetski pregled 2008; 65(9): 663-669.
29. Oliveira ACP, Teixeira CJ, Stefanello TF, Carrara MA, Bazotte RB, Sa-Nakanishi AB et al. Oxidative stress parameters as biomarkers of risk factor for diabetic foot among the patients with type 2 diabetes. Braz Arch Biol Technol 2014; 57(2): 223-227.
30. El-Wassef M, El-Saeed GS, El-Tokhy SE, Raslan HM, Tawfeek S, Siam I and Salem SI. Oxidative DNA damage in patients with type 2 diabetes mellitus. Diabetol Croat 2012; 41(4): 121-127.
31. West IC. Radicals and oxidative stress in diabetes. Diabetic Med 2000; 17(3): 171-180.
32. Rawi SM, Mourad IM and Sayed DA. Biochemical and changes in experimental diabetes before and after treatment with Mangifera indica Psidium guava extracts. Int J Pharm Bio Sci 2011; 2(2): 29-41.
33. Piwkowska A, Rogacka D, Audzeyenka I, Jankowski M, and Angielski, S. High glucose concentration affects the oxidantâ€antioxidant balance in cultured mouse podocytes. J Cell Biochem 2011; 112(6): 1661-1672.
34. Reddy SG, Kumar RG, Spurthi MK, Saraswati M and Rani SH. Oxidative stress and DNA damage in diabetic nephropathy. J Anal Bio-Sci 2013; 36(2).
35. Kasznicki J, Kosmalski M, Sliwinska A, Mrowicka M, Stanczyk M, Majsterek I and Drzewoski, J. Evaluation of oxidative stress markers in pathogenesis of diabetic neuropathy. Mol Biol Rep 2012; 39(9): 8669-8678.
36. Kaneto H, Fujii J, Suzuki K, Kasai H, Kawamori R, Kamada T and Taniguchi N. DNA cleavage induced by glycation of Cu, Zn-superoxide dismutase. Biochem J 1994; 304(1): 219-225.
37. Angeline T, Caroline JC, Rani JJA, Asirvatham AJ and Isabel W. Analysis of DNA damage, plasma lipid peroxidation, and antioxidant status in type 2 diabetes mellitus patients among a south Indian population. Int J Diab Dev Ctries 2015; 35(3): 491-495.
38. Arif M, Islam MR, Waise TMZ, Hassan F, Mondal SI and Kabir Y. DNA damage and plasma antioxidant indices in Bangladeshi type 2 diabetic patients. Diabetes Metab 2010; 36(1): 51-57.
39. Goodarzi MT, Navidi AA, Rezaei M and Babahmadiâ€Rezaei H. Oxidative damage to DNA and lipids: correlation with protein glycation in patients with type 1 diabetes. J Clin Lab Anal 2010; 24(2): 72-76.
40. Ibarra-Costilla E, Cerda-Flores RM, Davila-RodrÃguez MI, Samayo-Reyes A, Calzado-Flores C and Cortes-Gutierrez EI. DNA damage evaluated by comet assay in Mexican patients with type 2 diabetes mellitus. Acta Diabetol 2010; 47(1): 111-116.
41. Blasiak J, Arabski M, Krupa R, Wozniak K, Zadrozny M, Kasznicki J et al. DNA damage and repair in type 2 diabetes mellitus. Mutat Res Fundam Mol Mech Mutagen 2004; 554(1): 297-304.
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