Expression of 8-hydroxydeoxyguanosine in Nephrolithiatic Renal Tissues and Toxicity of Calcium Oxalate Monohydrate to Human Kidney Cell Line

Authors

  • Uraiwan Waiwijit Department of Biochemistry, Faculty of Medicine, Chulalongkorn University
  • Chanchai Boonla Department of Biochemistry, Faculty of Medicine, Chulalongkorn University
  • Wipawee Kitikowit Department of Pathology, Faculty of Medicine, Chulalongkorn University
  • Supoj Ratchanon Division of Urology, Department of Surgery, King Chulalongkorn Memorial Hospital
  • Sombat Borwornpadungkitti Division of urology, Department of Surgery, Khonkaen Hospital
  • Piyarattana Tosukhowong Department of Biochemistry, Faculty of Medicine, Chulalongkorn University

Keywords:

calcium oxalate monohydrate, oxidative stress, reactive oxygen species, 8-hydroxydeoxyguanosin

Abstract

Objective: To explore expression of 8-hydroxydeoxyguanosine (8-OHdG, a marker of oxidative DNA damage) in renal tissues obtained from calcium oxalate nephrolithiasis patients, and to investigate the effects of calcium oxalate monohydrate (COM) crystals on viability of human proximal renal tubular cells (HK-2) and intracellular reactive oxygen species (ROS) production.

Design: Observational analytical study.

Methods: Expression of 8-OHdG in paraffin-embedded nephrolithiatic renal sections (n=10) was explored by immunohistochemical staining. MTT assay was used to assess viability of HK-2 cells after exposed to COM in both dose and time-dependent experiments. The effect of COM on intracellular ROS production in HK-2 cells was determined using dichlorofluorescein method.

Results: In stone-containing renal tissues, 8-OHdG was strongly expressed in renal tubular cells. There was no expression of 8-OHdG in normal renal tissues. COM decreased viability of HK-2 cells in both dose- and time-dependent maners. Intracellular ROS generation was increased in HK-2 cells after treated with COM.

Conclusions: 8-OHdG was overexpressed in the kidneys of nephrolithiasis patients suggested that oxidative stress involved in the progress of kidney stone disease. COM was injurous to renal tubular cells. A decreased cell survival after treatment with COM may be mediated via ROS toxicity. Attenuation of oxidative damage by antioxidants is recommended for medical management of stone patietns.

References

Coe FL, Evan A, Worcester E. Kidney stone disease. Journal of Clinical Investigation 2005; 115(10): 2598-608.

Khan SR. Calcium oxalate crystal interaction with renal tubular epithelium, mechanism of crystal adhesion and its impact on stone development. Urological Research 1995; 23(2): 71-9.

Kohjimoto Y, Ebisuno S, Tamura M, Ohkawa T. Adhesion and endocytosis of calcium oxalate crystals on renal tubular cells. Scanning Microscopy 1996; 10(2): 459-70.

Schubert G. Stone analysis. Urological Research 2006; 34(2): 146-50.

McMartin KE, Wallace KB. Calcium oxalate monohydrate, a metabolite of ethylene glycol, is toxic for rat renal mitochondrial function. Toxicological Sciences 2005; 84(1): 195-200.

Hackett RL, Shevock PN, Khan SR. Madin-Darby canine kidney cells are injured by exposure to oxalate and to calcium oxalate crystals. Urological Research 1994; 22(4): 197-204.

Lahme S, Feil G, Strohmaier WL, Bichler KH, Stenzl A. Renal tubular alteration by crystalluria in stone disease - An experimental study by means of MDCK cells. Urologia Internationalis 2004; 72(3): 244-51.

Schepers MSJ, Van Ballegooijen ES, Bangma CH, Verkoelen CF. Crystals cause acute necrotic cell death in renal proximal tubule cells, but not in collecting tubule cells. Kidney International 2005; 68(4): 1543-53.

Rashed T, Menon M, Thamilselvan S. Molecular mechanism of oxalate-induced free radical production and glutathione redox imbalance in renal epithelial cells: Effect of antioxidants. American Journal of Nephrology 2004; 24(5): 557-68.

Habibzadegah-Tari P, Byer KG, Khan SR. Reactive oxygen species mediated calcium oxalate crystal-induced expression of MCP- 1 in HK-2 cells. Urological Research 2006; 34(1): 26-36.

Valavanidis A, Vlachogianni T, Fiotakis C. 8-hydroxy-2' -deoxyguanosine (8-OHdG): A critical biomarker of oxidative stress and carcinogenesis. Journal of environmental science and health Part C, Environmental carcinogenesis & ecotoxicology reviews 2009; 27(2): 120-39.

Pilger A, Rdiger HW. 8-Hydroxy-2-deoxyguanosine as a marker of oxidative DNA damage related to occupational and environmental exposures. International Archives of Occupational and Environmental Health 2006; 80(1): 1-15.

Seki S, Kitada T, Sakaguchi H, Nakatani K, Wakasa K. Pathological significance of oxidative cellular damage in human alcoholic liver disease. Histopathology 2003; 42(4): 365-71.

Boonla C, Wunsuwan R, Tungsanga K, Tosukhowong P. Urinary 8-hydroxydeoxyguanosine is elevated in patients with nephrolithiasis. Urological Research 2007; 35(4): 185-91.

Knoll T, Steidler A, Trojan L, Sagi S, Schaaf A, Yard B, et al. The influence of oxalate on renal epithelial and interstitial cells. Urological Research 2004; 32(4): 304-9.

Verkoelen CF, Schepers MSJ, Van Ballegooijen ES, Bangma CH. Effects of luminal oxalate or calcium oxalate on renal tubular cells in culture. Urological Research 2005; 33(5): 321-8.

Yao X, Zhong L. Genotoxic risk and oxidative DNA damage in HepG2 cells exposed to perfluorooctanoic acid. Mutation Research - Genetic Toxicology and Environmental Mutagenesis 2005; 587(1-2): 38-44.

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Published

2010-12-01

How to Cite

Waiwijit, U., Boonla, C., Kitikowit, W., Ratchanon, S., Borwornpadungkitti, S., & Tosukhowong, P. (2010). Expression of 8-hydroxydeoxyguanosine in Nephrolithiatic Renal Tissues and Toxicity of Calcium Oxalate Monohydrate to Human Kidney Cell Line. Insight Urology, 31(2), 137–146. Retrieved from https://he02.tci-thaijo.org/index.php/TJU/article/view/252407

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Original article