Investigation of the effects of lipoic acid and dihydrolipoate on experimental renal ischemia-reperfusion model

Abstract

Objective: Ischemic/reperfusion (I/R) causes tissue injury and the leading cause of acute kidney injury. In this study, we aimed to investigate the effects of the long and short-term usage of ALA and short-term DHLA on oxidative stress markers in the experimental renal ischemia-reperfusion model. Methods: Forty male rats (250 to 300 gr) were divided into 5 groups: control; I/R group; long-term ALA+IR group; short-term ALA+IR group; and short-term DHLA+IR group. Ischemia was carried out for 45 minutes followed by reperfusion for 4 hours. Thiobarbituric acid reactive sunstances (TBARM), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities in tissue samples and serum total antioxidant status (TAS) and total oxidative stress (TOS) assayed by the spectrophotometrically. Tissue samples were investigated by histopathological analyzes. Results: TBARM (Control: 0.38±0.05. I/R: 1.37±0.17, long-term ALA-treated group:1.025±0.15, short-term ALA-treated group: 0.68±0.09, short-term DHLA-treated group: 0.38±0.04 (nmol/mg protein); p<0,001) CAT (Control: 0.12±0.02, I/R: 0.04±0.008, long-term ALA-treated group: 0.07±0.01, short-term ALA-treated group:0.06±0.008, short-term DHLA-treated group: 0.08±0.01 (k/mg protein); p<0.001), GSH-Px (Control: 0.45±0.04, I/R: 0.21±0.028, long-term ALA-treated group: 0.37±0.05, short-term ALA-treated group :0.34±0.05, short-term DHLA-treated group: 0.37±0.04 (U/mg protein); p<0.001), and serum OSI levels (Control: 1.32±0.15, I/R: 3.08±0.44, long-term ALA-treated group: 1.775±0.21, short-term ALA-treated group: 1.85±0.37, short-term DHLA-treated group: 1.53±0.21 (arbitrary unit) ; p<0.001) were improved in the long and short-term ALA-treated group and short-term DHLA-treated group compared to the I/R group. These findings were more prominent in histopathological tissue samples in the DHLA-treated group. Conclusion: We consider that both long-term and short-term ALA applications have the potential for the treatment of renal I/R damage. Besides, DHLA is more effective than ALA.

Keywords

• Ischemic Damage
• Reperfusion
• Lipoic Acid
• Dihydrolipoat
• Antioxidant Enzyme

Project Number

16341

Deneysel böbrek iskemi-reperfüzyon modelinde lipoik asit ve dihidrolipoat kullanımının etkilerinin incelenmesi

Abstract

Amaç: İskemi/reperfüzyon (I/R) doku hasarına neden olarak akut böbrek hasarına yol açar. Bu çalışmada, deneysel böbrek iskemi-reperfüzyon modelinde uzun ve kısa süreli ALA ve kısa süreli DHLA kullanımının oksidatif stres belirteçleri üzerine etkilerinin araştırması amaçlanmıştır. . Yöntem: Kırk adet erkek rat (250-300 gr) 5 gruba ayrılmıştır: kontrol grubu; I/R grubu; uzun vadeli ALA+IR grubu; kısa vadeli ALA+IR grubu ve kısa vadeli DHLA+IR grubu. 45 dakika süreyle iskemi, ardından 4 saat süreyle reperfüzyon uygulanmıştır. Doku örneklerinde Tiyobarbitürik asid reaktif maddeler (TBARM), katalaz (CAT), süperoksit dismutaz (SOD) ve glutatyon peroksidaz (GSH-Px) aktiviteleri ile total antioksidan durum (TAS) ve total oksidatif stres kapasitesi (TOS) spektrofotometrik olarak ölçülmüştür. Doku örnekleri ayrıca histopatolojik olarak analiz edilmiştir. Bulgular: Uzun ve kısa süreli ALA uygulanan grupta ve kısa süreli DHLA uygulanan grupta I/R grubuna kıyasla TBARM (Kontrol: 0.38±0.05, I/R: 1.37±0.17, uzun vadeli ALA+IR grubu: 1.025±0.15, kısa vadeli ALA+IR grubu: 0.68±0.09, kısa vadeli DHLA+IR grubu: 0,38±0,04 (nmol/mg protein); p<0.001), CAT (Kontrol: 0.12±0.02, I/R: 0.04±0.008, uzun vadeli ALA+IR grubu: 0,07±0,01, kısa vadeli ALA+IR grubu: 0,06±0,008, kısa vadeli DHLA+IR grubu: 0.08±0.01 (k/mg protein; p<0.001), GSH-Px (Kontrol: 0.45±0.04, I/R: 0.21±0.028, uzun vadeli ALA+IR grubu: 0.37±0.05, kısa vadeli ALA+IR grubu: 0.34±0.05, kısa vadeli DHLA+IR grubu: 0.37±0.04 (U/mg protein); p<0.001), ve serum OSI (Kontrol: 1.32±0.15, I/R: 3.08±0.44, uzun vadeli ALA+IR grubu: 1.775±0.21, kısa vadeli ALA+IR grubu: 1.85±0.37, kısa vadeli DHLA+IR grubu: 1.53±0.21 (arbitary unit); p<0.001) seviyelerinde iyileşme gözlemlenmiştir. Bu bulgular, histopatolojik doku örneklerinde DHLA uygulanan grupta daha belirgindir Sonuç: Hem uzun süreli hem de kısa süreli ALA uygulamalarının renal I/R hasarının tedavisi için potansiyele sahip olduğu düşünülmektedir. Ayrıca DHLA’nın, ALA’dan daha etkili olduğu düşünülmektedir.

Keywords

• İskemik Hasar
• Reperfüzyon
• Lipoik Asit
• Dihidrolipoat
• Antioksidan Enzim

Supporting Institution

Hatay Mustafa Kemal University Scientific Research Project Coordination Unit (Hatay Mustafa Kemal Üniversitesi Bilimsel Araştırma Projeleri Birimi)

Project Number

16341

Thanks

Hatay Mustafa Kemal University Scientific Research Project Coordination Unit (Hatay Mustafa Kemal Üniversitesi Bilimsel Araştırma Projeleri Birimi)

References

1. Fan LH, He L, Cao ZQ, Xiang B, Liu L. Effect of ischemia preconditioning on renal ischemia/reperfusion injury in rats. Int Braz J Urol 2012; 38(6):842-54. https://doi.org/10.1590/1677-553820133806842.
2. Gomelsky A, Abreo K, Khater N, Abreo A, Amin B, Craig MK et al. Perioperative acute kidney injury: Stratification and risk reduction strategies. Best Pract Res Clin Anaesthesiol. 2020 Jun;34(2):167-182. https://doi.org/10.1016/j.bpa.2020.04.003.
3. Cheng J, Zhu P, Qin H, Li X, Yu H, Peng X. Dexmedetomidine attenuates cerebral ischemia/reperfusion injury in neonatal rats by inhibiting TLR4 signaling. J Int Med Res 2018; 46(7):2925-32. https://dx.doi.org/10.1177%2F0300060518781382.
4. Ozcan O, Erdal H, Cakirca G, Yonden Z. Oxidative stress and its impacts on intracellular lipids, proteins and DNA. JCEI 2015 ;6(3):331-6. https://doi.org/10.5799/ahinjs.01.2015.03.0545.
5. Nath KA, Norby SM. Reactive oxygen species and acute renal failure. Am J Med 2000; 109(8):665-78. https://doi.org/10.1016/s0002-9343(00)00612-4.
6. Ergün O, Ulman C, Kilicalp AS, Ulman I. Carnitine as a preventive agent in experimental renal ischemia-reperfusion injury. Urol Res 2001; 29(3):186-9. https://doi.org/10.1007/s002400100176.
7. Oruc O, Inci K, Aki FT, Zeybek D, Muftuoglu SF, Kilinc K et al. Sildenafil attenuates renal ischemia reperfusion injury by decreasing leukocyte infiltration. Acta Histochem. 2010 Jul;112(4):337-44. https://doi.org/10.1016/j.acthis.2009.02.005.
8. Skibska B, Goraca A. The protective effect of lipoic acid on selected cardiovascular diseases caused by age-related oxidative stress. Oxid Med Cell Longev 2015; 2015:313021. https://doi.org/10.1155/2015/313021.

9. Al-Rasheed NM, Attia HA, Hasan IH, Al-Amin M, Al-Ajmi H, Mohamad RA. Adverse cardiac responses to alpha-lipoic acid in a rat-diabetic model: possible mechanisms? J Physiol Biochem 2013; 69(4):761-78. https://doi.org/10.1007/s13105-013-0252-9.
10. Bae EH, Lee KS, Lee J, Ma SK, Kim NH, Choi KC et al. Effects of alpha-lipoic acid on ischemia-reperfusion-induced renal dysfunction in rats. Am J Physiol Renal Physiol. 2008 Jan;294(1):F272-80. https://doi.org/10.1152/ajprenal.00352.2007.
11. Karaca EG. Lipoik Asit: Evrensel Antioksidan. Afyon Kocatepe Üniversitesi Fen Bilimleri Dergisi 2009; 8(1):231-46. https://dergipark.org.tr/tr/pub/akufemubid/issue/1607/20056.
12. Kurutas EB. The importance of antioxidants which play the role in cellular response against oxidative/nitrosative stress: current state. Nutr J 2016; 15(1):1-22. https://doi.org/10.1186/s12937-016-0186-5.
13. Navari IF, Frank QM, Cristina S. Lipoic acid: a unique antioxidant in the detoxification of activated oxygen species. Plant Physiology and Biochemistry 2002; 40(6-8):463-70. https://doi.org/10.1016/S0981-9428(02)01407-9.
14. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248-54. https://doi.org/10.1006/abio.1976.9999.
15. Hammouda A el-R, Khalil MM, Salem A. Lipid peroxidation products in pleural fluid for separation of transudates and exudates. Clin Chem 1995; 41(9):1314-5. https://doi.org/10.1093/clinchem/41.9.1314.
16. Aebi H. Catalase. Catalase in vitro. In: Methods in enzymology. Academic Press, 1984: 121-6. https://doi.org/10.1016/s0076-6879(84)05016-3.
17. Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 1967; 70(1):158-69. https://doi.org/10.5555/uri:pii:0022214367900765.
18. Durak I, Isık CU, Canbolat O, Akyol O, Kavutcu M. Adenosine deaminase, 5′ nucleotidase, xanthine oxidase, superoxide dismutase, and catalase activities in cancerous and noncancerous human laryngeal tissues. J Free Rad Biol Med 1993; 15(6):681-4. https://doi.org/10.1016/0891-5849(93)90174-s.
19. Erel O. A new automated colorimetric method for measuring total oxidant status. Clin Biochem 2005; 38(12):1103-11. https://doi.org/10.1016/j.clinbiochem.2005.08.008.
20. Goujon JM, Hauet T, Menet E, Levillain P, Babin P, Carretier M. Histological evaluation of proximal tubule cell injury in isolated perfused pig kidneys exposed to cold ischemia. J Surg Res 1999; 82(2):228-33. https://doi.org/10.1006/jsre.1998.5526.
21. Takaoka M, Ohkita M, Kobayashi Y, Yuba M, Matsumura Y. Protective effect of alpha-lipoic acid against ischaemic acute renal failure in rats. Clin Exp Pharmacol Physiol 2002; 29(3):189-94. https://doi.org/10.1046/j.1440-1681.2002.03624.x.
22. Catalá A. An overview of lipid peroxidation with emphasis in outer segments of photoreceptors and the chemiluminescence assay. Int J Biochem Cell Biol 2006; 38(9):1482-95. https://doi.org/10.1016/j.biocel.2006.02.010.
23. Cosar E, Sahin FK, Koken G, Toy H, Basarali K, Büyükbas S. The protective effect of alpha-lipoic acid in experimental ovarian ischaemia-reperfusion injury. Aust N Z J Obstet Gynaecol 2007; 47(6):499-503. https://doi.org/10.1111/j.1479-828x.2007.00787.x.
24. Sehirli O, Sener E, Cetinel S, Yüksel M, Gedik N, Sener G. Alpha-lipoic acid protects against renal ischaemia-reperfusion injury in rats. Clin Exp Pharmacol Physiol 2008; 35(3):249-55. https://doi.org/10.1111/j.1440-1681.2007.04810.x.
25. Wongmekiat O, Leelarungrayub D, Thamprasert K. Alpha-lipoic acid attenuates renal injury in rats with obstructive nephropathy. Biomed Res Int 2013; 2013:138719. https://doi.org/10.1155/2013/138719.
26. Gorąca A, Huk-Kolega H, Piechota A, Kleniewska P, Ciejka E, Skibska B. Lipoic acid - biological activity and therapeutic potential. Pharmacol Rep 2011; 63(4):849-58. https://doi.org/10.1016/s1734-1140(11)70600-4.
27. Handelman GJ, Han D, Tritschler H, Packer L. Alpha-lipoic acid reduction by mammalian cells to the dithiol form, and release into the culture medium. Biochem Pharmacol 1994; 47(10):1725-30. https://doi.org/10.1016/0006-2952(94)90298-4.
28. Packer L, Witt EH, Tritschler HJ. alpha-Lipoic acid as a biological antioxidant. J Free Rad Biol Med. 1995; 19(2):227-50. https://doi.org/10.1016/0891-5849(95)00017-r.
29. Sivaprasad R, Nagaraj M, Varalakshmi P. Combined efficacies of lipoic acid and 2,3-dimercaptosuccinic acid against lead-induced lipid peroxidation in rat liver. J Nutr Biochem 2004; 15(1):18-23. https://doi.org/10.1016/j.jnutbio.2003.09.001.
30. Kagan VE, Shvedova A, Serbinova E, Khan S, Swanson C, Powell R et al. Dihydrolipoic acid--a universal antioxidant both in the membrane and in the aqueous phase. Reduction of peroxyl, ascorbyl and chromanoxyl radicals. Biochem Pharmacol. 1992 Oct 20;44(8):1637-49. https://doi.org/10.1016/0006-2952(92)90482-x.
31. Bast A, Haenen GR. The toxicity of antioxidants and their metabolites. Environ Toxicol Pharmacol 2002; 11(3-4):251-8. https://doi.org/10.1016/s1382-6689(01)00118-1.
32. Dobashi K, Ghosh B, Orak JK, Singh I, Singh AK. Kidney ischemia-reperfusion: modulation of antioxidant defenses. Mol Cell Biochem 2000; 205(1-2):1-11. https://doi.org/10.1023/a:1007047505107.
33. Williams P, Lopez H, Britt D, Chan C, Ezrin A, Hottendorf R. Characterization of renal ischemia-reperfusion injury in rats. J Pharmacol Toxicol Methods 1997; 37(1):1-7. https://doi.org/10.1016/s1056-8719(96)00141-4.
34. Hussein AA, Barakat N, Awadalla A, Shokeir AA. Systemic and renal haemodynamic changes in renal schemia/reperfusion injury: impact of erythropoietin. Can J Physiol Pharmacol 2012; 90(11):1535-43. https://doi.org/10.1139/y2012-120.