Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
권호 표지

Protective effects of EDTA and EGTA against CCl4-induced acute hepatotoxicity in mice

마우스에서 사염화탄소로 유발된 급성 간독성에 대한 EDTA 및 EGTA의 보호효과

  • Park, Seung-Guk (School of Veterinary Medicine, Kangwon National University (Institute of Veterinary Science)) ;
  • Cho, Yong-Do (School of Veterinary Medicine, Kangwon National University (Institute of Veterinary Science)) ;
  • Shin, Taekyun (Department of Veterinary Medicine, Chueju National University) ;
  • Wie, Myung-Bok (School of Veterinary Medicine, Kangwon National University (Institute of Veterinary Science))
  • 박승국 (강원대학교 수의학부대학 (동물의학종합연구소)) ;
  • 조용도 (강원대학교 수의학부대학 (동물의학종합연구소)) ;
  • 신태균 (제주대학교 수의학과) ;
  • 위명복 (강원대학교 수의학부대학 (동물의학종합연구소))
  • Accepted : 2007.08.30
  • Published : 2007.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study investigated the protective effects of ethylene glycol-bis(${\beta}$-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), an extracellular calcium chelator, and ethylenediaminetetraacetic acid (EDTA), which chelates calcium and most metal ions, against carbon tetrachloride ($CCl_4$)-induced acute hepatotoxicity in mice. Mice were treated with EGTA or EDTA at a dose of 20 (low) or 100 mg/kg (high) subcutaneously 1h before $CCl_4$ administration. The mice were fasted and sacrificed 18h after $CCl_4$ treatment. Blood samples were collected from the carotid artery by decapitation under light ether anesthesia. Serum alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and cholesterol levels were measured. Malondialdehyde (MDA) production was determined as an index of lipid peroxidation in the liver. The liver, kidneys, and spleen were weighed. We also evaluated the histopathological changes in the liver in each group. The relative weights of the liver were significantly higher in the $CCl_4$-treatment group than in the normal group, except in the high-EDTA treatment group. EGTA and EDTA treatment caused a significant decrease in serum ALP, ALT, and AST levels. Of all of the doses of EGTA and EDTA tested, the high-EDTA dose resulted in the most remarkable inhibitory action. The protective effect in the high-EDTA-treatment group was confirmed histopathologically. The low-EGTA-treatment group showed a significant decrease in serum TG and cholesterol levels. Liver MDA levels were significantly decreased in the EGTA (20 mg/kg) and EDTA (20, 100 mg/kg) groups. These results suggest that EDTA, which chelates both calcium and metal ions, confers better protection in $CCl_4$-induced acute liver damage than does EGTA, a calcium chelator.

Keywords

References

  1. 배지혜, 윤종국, 이상일. 흰쥐에 있어서 사염화탄소에 의한 간손상의 allopurinol의 영향. 한국독성학회지. 1995, 11, 247-252
  2. Agarwal AK, Mehendale HM. Excessive hepatic accumulation of intracellular Ca$^{2+}$ in chlordecone potentiated CCl$_{4}$ toxicity. Toxicology 1984, 30, 17-24 https://doi.org/10.1016/0300-483X(84)90058-1
  3. Albano E, Carini R, Parola M, Bellomo G, GoriaGatti L, Poli G, Dianzani MU. Effects of carbon tetrachloride on calcium homeostasis. A critical reconsideration. Biochem Pharmacol 1989, 38, 2719-2725 https://doi.org/10.1016/0006-2952(89)90559-5
  4. Boullerne AI, Nedelkoska L, Benjamins JA. Role of calcium in nitric oxide-induced cytotoxicity: EGTA protects mouse oligodendrocytes. J Neurosci Res 2001, 63, 124-135 https://doi.org/10.1002/1097-4547(20010115)63:2<124::AID-JNR1004>3.0.CO;2-C
  5. Cherny RA, Legg JT, McLean CA, Fairlie DP, Huang X, Atwood CS, Beyreuther K, Tanzi RE, Masters CL, Bush AI. Aqueous dissolution of Alzheimer's disease A$\beta$amyloid deposits by biometal depletion. J Biol Chem 1999, 274, 23223-23228 https://doi.org/10.1074/jbc.274.33.23223
  6. de Ferreyra EC, Bernacchi AS, Villarruel MC, de Fenos OM, Castro JA. Prevention of CC14-induced liver necrosis by the calcium chelator arsenazo III. Exp Mol Pathol 1993, 58, 194-204 https://doi.org/10.1006/exmp.1993.1017
  7. Ekstrom G, Ingelman-Sundberg M. Mechanisms of lipid peroxidation dependent upon cytochrome P-450 LM$_{2}$. Em J Biochem 1986, 158, 195-201 https://doi.org/10.1111/j.1432-1033.1986.tb09738.x
  8. Fisher AEO, Maxwell SC, Naughton DP. Superoxide and hydrogen peroxide suppression by metal ions and their EDTA complexes. Biochem Biophys Res Commun 2004, 316, 48-51 https://doi.org/10.1016/j.bbrc.2004.02.013
  9. Frezza EE, Gerunda GE, Farinati F, DeMaria N, Galligioni A, Plebani F, Giacomin A, Van Thiel DH. CCl$_{4}$-induced liver cirrhosis and hepatocellular carcinoma in rats: relationship to plasma zinc, copper and estradiol levels. Hepatogastroenterology 1994, 41, 367-369
  10. Halliwell B, Gutteridge JM. The importance of free radicals and catalytic metal ions in human diseases. Mol Aspects Med 1985, 8, 89-193 https://doi.org/10.1016/0098-2997(85)90001-9
  11. Heindorff K, Aurich O, Michaelis A, Rieger R. Genetic toxicology of ethylenediaminetetraacetic acid (EDTA). Mutat Res 1983, 115, 149-173 https://doi.org/10.1016/0165-1110(83)90001-5
  12. Klaassen CD. Nonmetallic environmental toxicants: air pollutants, solvents and vapors, and pesticides. In: Goodman LS, Gilman AG (eds.). The pharmacological basis of therapeutics. 8th ed. pp. 1622-1623, Pergamon Press, New York, 1985
  13. Mansour MA. Protective effects of thymoquinone and desferrioxamine against hepatotoxicity of carbon tetrachloride in mice. Life Sci 2000, 66, 2583-2591 https://doi.org/10.1016/S0024-3205(00)00592-0
  14. McCay PB, Lai EK, Poyer JL, DuBose CM, Janzen EG. Oxygen and carbon-centered free radical formation during carbon tetrachloride metabolism. Observation of lipid radicals in vivo and in vitro. J Biol Chem 1993, 259, 2135-2143
  15. Minotti G. Aust SD. The role of iron in oxygen radical mediated lipid peroxidation. Chem Biol Interact 1989, 71, 1-19 https://doi.org/10.1016/0009-2797(89)90087-2
  16. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979, 95, 351-358 https://doi.org/10.1016/0003-2697(79)90738-3
  17. Younes M, Siegers CP. The role of iron in the paracetamol- and CCl$_{4}$-induced lipid peroxidation and hepatotoxicity. Chem Biol Interact 1985, 55, 327-334 https://doi.org/10.1016/S0009-2797(85)80139-3