IN VITRO EVALUATION OF THE EFFECT OF GLUTATHIONE ON CASPASE SYSTEM AND OXIDATIVE DNA DAMAGE IN HIGH GLUCOSE CONDITION

Yıl 2021, Cilt: 10 Sayı: 2, 132 - 137, 30.07.2021

Gülay Yaycı Semiha Dede , Ayşe Usta , Veysel Yüksek , Sedat Çetin

https://doi.org/10.18036/estubtdc.802794

https://izlik.org/JA97DE52ZN

Öz

This study was planned to investigate the effects of glutathione, known to be a strong antioxidant, on caspase-dependent apoptosis and oxidative DNA damage in the kidney cells (BHK-21) exposed to high glucose. BHK-21 cell line was cultivated by regular passages in vitro conditions. Study groups were set up as control, study and its combinations groups ((Glucose; (285 mM, HG), glutathione (250 µM)). After 24 hours of incubation, trypsinized cells were disrupted by freeze-thaw method and analyte was prepared. Caspase 3, 8, 9, M30 and 8-OHdG (oxidative DNA damage marker) levels were determined using commercial ELISA kits. M30, caspase 3, 8 and 9 which are the parameters of apoptosis, were found highest in the HG group. In GSH-treated groups, these parameters decreased slightly. There was no significant difference in the levels of 8-OHdG which is the indicator of oxidative DNA damage.

Anahtar Kelimeler

High glucose , glutathione , apoptosis , DNA damage , in vitro , kidney cells

Destekleyen Kurum

Van Yüzüncü yıl üniversitesi

Proje Numarası

2015-SBE-YL208

Kaynakça

• [1] Abou-Seif MA, Youssef AA. Evaluation of some biochemical changes in diabetic patients. Clin Chim Acta, 2004;346(2):161-170.
• [2] Andian G, Burçak G. Oxidative damage to nuclear DNA in streptozotocin-diabetic rat liver. Clin Exp Pharmacol Physiol, 2005;32(8):663-666.
• [3] ADA (American Diabetes Association). Diagnosis and classification of diabetes mellitus. Diabet Care, 2011;34(1):62-69.
• [4] Bildik A, Yur F, H Camas, Dede S. Investigation of glutathione and lipid peroxidation levels with respect to haemoglobin types in lambs with white muscle disease. Van Vet J, 1996;7(1-2):95-98.
• [5] Franco R, Schoneveld OJ, Pappa A, Panayiotidis MI. The central role of glutathione in the pathophysiology of human diseases. Arch Physiol Biochem, 2007;113:243-258.
• [6] Dede S, Çamaş H. Determination of some biochemical parameters (GSH, Hb and Tf types and Mn) in laying hens and study its effects on egg production. YYU Sag Bil Derg, 2001;7:131-138.
• [7] Ulukaya E, Pirianov G, Kurt MA, Wood EJ, Mehmet H. Fenretinide induces cytochrome C release, caspase 9 activation and apoptosis in the absence of mitochondrial membrane depolarisation. Cell Death Different, 2003;10:856-859.
• [8] Danial NN, Korsmeyer SJ. Cell death: critical control points. Cell, 2004;11:205-219.
• [9] Elmore S. Apoptosis: A review of programmed cell death. Toxicol Pathol, 2007;35:495-516.
• [10] Schultz-Johanssen J, Harris AK, Rychly DJ, Ergul A. Oxidative stress and the use of antioxidants in diabetes. linking basic science to practice. Cardiovasc Diabetol, 2005;4-5.
• [11] Yur F, Dede S, Karaca T, CiftçiYegin S, Değer Y, Ozdemir H. The effect of glutathione treatment on the biochemical and immunohistochemical profile in streptozotocin-induced diabetic rats. J Membrane Biol, 2013;246(6):427-433.
• [12] Yur F, Dede S, Taşpınar M, Usta A, Cetin S. Effects of glutathione application on oxidative DNA damage and antioxidant system in the glucose-treated kidney cell line. YYU BAP, (unpublished data), Project No:2013-VF-B062.
• [13] Aksoy Y. The role of glutathione in antioxidant mechanism. Turk J Med Sci, 2002; 2(4):442-448.
• [14] Bonnefont-Rousselot D. Glucose and reactive oxygen species. Curr Opin Clin Nutr Metab Care, 2002;5:561-568.
• [15] Sekhar RV, McKay SV, Patel SG, Guthikonda AP, Reddy VT, Balasubramanyam A et al. Glutathione synthesis is diminished in patients with uncontrolled diabetes and restored by dietary supplementation with cysteine and glycine. Diabetes Care, 2011;34(1):162-167.
• [16] Kuchake VG, Upasani CD. Effect of Vitamin E and C plus reduced glutathione in treatment of diabetic nephropathy. Int J Pharm Sci Rev Res, 2013;2(12):1-5.
• [17] Sahin E, Gocmen AY, Koçak, Tuncer M, Gumuslu S. The association of advanced glycation end-products with glutathione status. Ann Clin Biochem, 2008;45(Pt 4):369-374.
• [18] Tan H, Qurashi A, Poidevin M, Nelson DL, Li H, Jin P. Retrotransposon activation contributes to fragile X premutation RCGG-mediated neurodegeneration. Hum Mol Genet, 2012;21(1):57-65.
• [19] Schutte B, Henfling M, Kölgen W, Bouman M, Meex S, Leers MPG et al. Keratin 8/18 break down and reorganization during apoptosis. Exp Cell Res, 2004;297(1):11-26.
• [20] Prabhakar S, Starnes J, Shi S, Lonis B, Tran R. Diabetic nephropathy is associated with oxidative stress and decreased renal nitric oxide production. J Am Soc Nephrol, 2007;18(11):2945-2952.
• [21] Park A, Suzuki T, Lennarz WJ. Identification of proteins that interact with mammalian peptide: n-glycanase and implicate this hydrolase in the proteasome-dependent pathway for protein degradation. Proc Nation Acad Sci, 2001;98(20):11163-11168.
• [22] Ueno Y, Kizaki M, Nakagiri RKamiya T, Sumi H, Osawa T. Dietary glutathione protects rats from diabetic nephropathy and neuropathy. J Nutr 2002;132(5):897-900.