Chonnam Med J.  2010 Dec;46(3):156-162. 10.4068/cmj.2010.46.3.156.

Effects of Antioxidants in Cisplatin-Induced Renal Tubular Apoptosis

• Bae EH
• Lee J
• Ma SK
• Kim SW

• KMID: 2116769
• DOI: http://doi.org/10.4068/cmj.2010.46.3.156

Abstract

The present study was designed to evaluate the possible renoprotective effects of alpha-lipoic acid (alpha-LA) in cisplatin-induced nephropathy and its role in inflammation and apoptosis in the kidney. Male Sprague-Dawley rats, weighing 180~200 g, were used. One group of rats was injected with cisplatin (6 mg/kg intraperitoneally). Another group of rats was injected with cisplatin and co-treated with alpha-LA. Rats that received an injection of vehicle only served as controls. Four days later, the mRNA expression of Bax and Bcl-2 was determined by real-time PCR. The protein expression of ED-1, cyclooxygenase-2 (COX2), heat shock protein 72 (HSP72), Bax, and Bcl-2 was determined in the kidney by immunoblotting. Apoptosis was determined by TUNEL staining. Cisplatin-treated rats had decreased creatinine clearance and increased plasma creatinine levels compared with controls. alpha-LA treatment lowered plasma creatinine levels and increased creatinine clearance. In cisplatin-treated rats, the mRNA expression of Bcl-2 was decreased, which was attenuated by alpha-LA treatment. The protein expression of ED-1, COX 2, HSP72, cleaved caspase-3, and Bax in the kidney was increased compared with controls, whereas that of Bcl-2 was decreased, and these changes were counteracted by alpha-LA treatment. TUNEL-positive cells were increased, in association with an increased protein expression of Bax and cleaved caspase-3 in the kidney of cisplatin-induced nephropathy, which was counteracted by alpha-LA treatment. These findings suggest that alpha-LA is effective in preventing the progression of cisplatin-induced nephropathy, the mechanism of which is associated with anti-inflammation and anti-apoptotic effects.

Keyword

Oxidative stress; Cisplatin; Apoptosis; Inflammation

MeSH Terms

Animals
Antioxidants
Apoptosis
Caspase 3
Cisplatin
Creatinine
Cyclooxygenase 2
HSP72 Heat-Shock Proteins
Humans
Immunoblotting
In Situ Nick-End Labeling
Inflammation
Kidney
Male
Oxidative Stress
Plasma
Rats
Rats, Sprague-Dawley
Real-Time Polymerase Chain Reaction
RNA, Messenger
Thioctic Acid
Antioxidants
Caspase 3
Cisplatin
Creatinine
Cyclooxygenase 2
HSP72 Heat-Shock Proteins
RNA, Messenger
Thioctic Acid

Figure

• 

  Fig. 1 Effects of alpha lipoic acid (α-LA) on serum creatinine level and creatinine clearance. *p<0.05 compared with control. †p<0.05 compared with cisplatin-treated rats.

• 

  Fig. 2 Effects of alpha lipoic acid (α-LA) on the protein expression of ED-1, COX2 and HSP70 in the kidney. *p<0.05 compared with control. †p<0.05 compared with cisplatin-treated rats.

• 

  Fig. 3 Effects of alpha lipoic acid (α-LA) on mRNA expression of Bax and Bcl-2 in the kidney. *p<0.05 compared with control. †p<0.05 compared with cisplatin-treated rats.

• 

  Fig. 4 Effects of alpha lipoic acid (α-LA) on the protein expression of Bax, Bcl-2 and cleaved caspase 3 in the kidney. *p<0.05 compared with control. †p<0.05 compared with cisplatin-treated rats.

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  Fig. 5 Apoptosis evaluated by TUNEL staining in the kidney sections from different groups of rats. The dark brown dots represent TUNEL-positive nuclei. G: glomerulus.

Reference

1. Einhorn LH, Williams SD. The role of cis-platinum in solid-tumor therapy. N Engl J Med. 1979. 300:289–291.

2. Einhorn LH, Donohue J. Cis-diamminedichloroplatinum, vinblastine, and bleomycin combination chemotherapy in disseminated testicular cancer. Ann Intern Med. 1977. 87:293–298.

3. Rozencweig M, von Hoff DD, Slavik M, Muggia FM. Cisdiamminedichloroplatinum (II). A new anticancer drug. Ann Intern Med. 1977. 86:803–812.

4. Arany I, Safirstein RL. Cisplatin nephrotoxicity. Semin Nephrol. 2003. 23:460–464.

5. Kim SW, Lee JU, Nah MY, Kang DG, Ahn KY, Lee HS, et al. Cisplatin decreases the abundance of aquaporin water channels in rat kidney. J Am Soc Nephrol. 2001. 12:875–882.

6. Li Q, Bowmer CJ, Yates MS. Effect of arginine on cisplatin-induced acute renal failure in the rat. Biochem Pharmacol. 1994. 47:2298–2301.

7. Pabla N, Dong Z. Cisplatin nephrotoxicity: mechanisms and renoprotective strategies. Kidney Int. 2008. 73:994–1007.

8. Pabla N, Dong Z. Cisplatin nephrotoxicity: mechanisms and renoprotective strategies. Kidney Int. 2008. 73:994–1007.

9. El Midaoui A, de Champlain J. Prevention of hypertension, insulin resistance, and oxidative stress by alpha-lipoic acid. Hypertension. 2002. 39:303–307.

10. Lee KM, Park KG, Kim YD, Lee HJ, Kim HT, Cho WH, et al. Alpha-lipoic acid inhibits fractalkine expression and prevents neointimal hyperplasia after balloon injury in rat carotid artery. Atherosclerosis. 2006. 189:106–114.

11. Sung MJ, Kim W, Ahn SY, Cho CH, Koh GY, Moon SO, et al. Protective effect of alpha-lipoic acid in lipopolysaccharide-induced endothelial fractalkine expression. Circ Res. 2005. 97:880–890.

12. Salahudeen A, Poovala V, Parry W, Pande R, Kanji V, Ansari N, et al. Cisplatin induces N-acetyl cysteine suppressible F2-isoprostane production and injury in renal tubular epithelial cells. J Am Soc Nephrol. 1998. 9:1448–1455.

13. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001. 25:402–408.

14. Davis CA, Nick HS, Agarwal A. Manganese superoxide dismutase attenuates Cisplatin-induced renal injury: importance of superoxide. J Am Soc Nephrol. 2001. 12:2683–2690.

15. Faubel S, Lewis EC, Reznikov L, Ljubanovic D, Hoke TS, Somerset H, et al. Cisplatin-induced acute renal failure is associated with an increase in the cytokines interleukin (IL)-1beta, IL-18, IL-6, and neutrophil infiltration in the kidney. J Pharmacol Exp Ther. 2007. 322:8–15.

16. Kaushal GP, Kaushal V, Hong X, Shah SV. Role and regulation of activation of caspases in cisplatin-induced injury to renal tubular epithelial cells. Kidney Int. 2001. 60:1726–1736.

17. Wang J, Wei Q, Wang CY, Hill WD, Hess DC, Dong Z. Minocycline up-regulates Bcl-2 and protects against cell death in mitochondria. J Biol Chem. 2004. 279:19948–19954.

18. Bhatt K, Feng L, Pabla N, Liu K, Smith S, Dong Z. Effects of targeted Bcl-2 expression in mitochondria or endoplasmic reticulum on renal tubular cell apoptosis. Am J Physiol Renal Physiol. 2008. 294:F499–F507.

19. He H, Lam M, McCormick TS, Distelhorst CW. Maintenance of calcium homeostasis in the endoplasmic reticulum by Bcl-2. J Cell Biol. 1997. 138:1219–1228.

20. Lithgow T, van Driel R, Bertram JF, Strasser A. The protein product of the oncogene bcl-2 is a component of the nuclear envelope, the endoplasmic reticulum, and the outer mitochondrial membrane. Cell Growth Differ. 1994. 5:411–417.

21. Hamet P, Malo D, Tremblay J. Increased transcription of a major stress gene in spontaneously hypertensive mice. Hypertension. 1990. 15:904–908.

22. Komatsuda A, Wakui H, Oyama Y, Imai H, Miura AB, Itoh H, et al. Overexpression of the human 72 kDa heat shock protein in renal tubular cells confers resistance against oxidative injury and cisplatin toxicity. Nephrol Dial Transplant. 1999. 14:1385–1390.

23. Ramesh G, Reeves WB. TNF-alpha mediates chemokine and cytokine expression and renal injury in cisplatin nephrotoxicity. J Clin Invest. 2002. 110:835–842.

24. Ramesh G, Brian Reeves W. Cisplatin increases TNF-alpha mRNA stability in kidney proximal tubule cells. Ren Fail. 2006. 28:583–592.

25. Zhang B, Ramesh G, Norbury CC, Reeves WB. Cisplatin-induced nephrotoxicity is mediated by tumor necrosis factor-alpha produced by renal parenchymal cells. Kidney Int. 2007. 72:37–44.

26. Yamate J, Tatsumi M, Nakatsuji S, Kuwamura M, Kotani T, Sakuma S. Immunohistochemical observations on the kinetics of macrophages and myofibroblasts in rat renal interstitial fibrosis induced by cisdiamminedichloroplatinum. J Comp Pathol. 1995. 112:27–39.

27. Adams JM, Cory S. The Bcl-2 protein family: arbiters of cell survival. Science. 1998. 281:1322–1326.