Effect of Ca2+ Channel Blockers, External Ca2+ and Phospholipase A2 Inhibitors on T-butylhydroperoxide-induced Lipid Peroxidation and Toxicity in Rat Liver Slices

Overview

Journal Korean J Intern Med

Specialty General Medicine

Date 1997 Jun 1

PMID 9439155

Citations 1

Authors

J Heo

G H Kim

K S Lee

W U Go

H J Ju

S K Park

C S Song

G A Song

M Cho

U S Yang

H K Moon

Y K Kim

Affiliations

Soon will be listed here.

Abstract

Objectives: This study was undertaken to examine the effect of oxidant on lipid peroxidation and lethal cell injury in rat liver slices.

Methods: t-Butylhydroperoxide (t-BHP) was employed as a model of an oxidant. The lipid peroxidation and lethal cell injury were estimated by measuring the formation of malondialdehyde (MDA) and lactate dehydrogenase (LDH) release, respectively.

Results: t-BHP increased lipid peroxidation and LDH release in a dose-dependent manner over concentrations of 0.5-10 mM. t-BHP-induced lipid peroxidation was completely prevented by an antioxidant, N,N-diphenyl-p-phenylenediamine (DPPD), but LDH release was partially decreased. Both t-BHP-induced lipid peroxidation and LDH release were significantly protected by iron chelator, deferoxamine, sulfhydryl reducing agent, dithiothreitol and glutathione. Ca2+ channel blockers, verapamil, diltiazem and nifedipine exerted a significant protective effect against t-BHP-induced lipid peroxidation and LDH release. By contrast, addition of external Ca2+ chelator, ethylene glycol bis(b-aminoethyl ether)-N,N-tetraacetic acid (EGTA) did not alter t-BHP-induced lipid peroxidation, whereas t-BHP-induced lethal cell injury was significantly prevented. Phospholipase A2 (PLA2) inhibitors, mepacrine and butacaine produced a partial protective effect.

Conclusions: These results suggest that t-BHP induces cell injury by lipid peroxidation-dependent and -independent mechanisms which can be partially prevented by Ca2+ channel blockers and PLA2 inhibitors.

Citing Articles

The protection of hepatocyte cells from the effects of oxidative stress by treatment with vitamin E in conjunction with DTT.

Tsai J, Chen H, Chen Y, Liu J, Lii C J Biomed Biotechnol. 2010; 2010:486267.

PMID: 20490359 PMC: 2872767. DOI: 10.1155/2010/486267.

References

Starke P, Hoek J, Farber J . Calcium-dependent and calcium-independent mechanisms of irreversible cell injury in cultured hepatocytes. J Biol Chem. 1986; 261(7):3006-12. View

Pereira R, Bertocchi A, Vercesi A . Protective effect of trifluoperazine on the mitochondrial damage induced by Ca2+ plus prooxidants. Biochem Pharmacol. 1992; 44(9):1795-801. DOI: 10.1016/0006-2952(92)90074-s. View

Bellomo G, Thor H, Orrenius S . Increase in cytosolic Ca2+ concentration during t-butyl hydroperoxide metabolism by isolated hepatocytes involves NADPH oxidation and mobilization of intracellular Ca2+ stores. FEBS Lett. 1984; 168(1):38-42. DOI: 10.1016/0014-5793(84)80202-1. View

Mihara M, Uchiyama M . Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem. 1978; 86(1):271-8. DOI: 10.1016/0003-2697(78)90342-1. View

Masaki N, Kyle M, Farber J . tert-butyl hydroperoxide kills cultured hepatocytes by peroxidizing membrane lipids. Arch Biochem Biophys. 1989; 269(2):390-9. DOI: 10.1016/0003-9861(89)90122-7. View

Trump B, Berezesky I . Calcium-mediated cell injury and cell death. FASEB J. 1995; 9(2):219-28. DOI: 10.1096/fasebj.9.2.7781924. View

Imberti R, Nieminen A, Herman B, Lemasters J . Mitochondrial and glycolytic dysfunction in lethal injury to hepatocytes by t-butylhydroperoxide: protection by fructose, cyclosporin A and trifluoperazine. J Pharmacol Exp Ther. 1993; 265(1):392-400. View

Bunnachak D, Almeida A, Wetzels J, Gengaro P, Nemenoff R, BURKE T . Ca2+ uptake, fatty acid, and LDH release during proximal tubule hypoxia: effects of mepacrine and dibucaine. Am J Physiol. 1994; 266(2 Pt 2):F196-201. DOI: 10.1152/ajprenal.1994.266.2.F196. View

Deakin C, Fagan E, Williams R . Cytoprotective effects of calcium channel blockers. Mechanisms and potential applications in hepatocellular injury. J Hepatol. 1991; 12(2):251-5. DOI: 10.1016/0168-8278(91)90947-a. View

10.

Katz A . Basic cellular mechanisms of action of the calcium-channel blockers. Am J Cardiol. 1985; 55(3):2B-9B. DOI: 10.1016/0002-9149(85)90607-1. View

11.

Freeman B, Crapo J . Biology of disease: free radicals and tissue injury. Lab Invest. 1982; 47(5):412-26. View

12.

POGGIOLI J, Mauger J, Guesdon F, Claret M . A regulatory calcium-binding site for calcium channel in isolated rat hepatocytes. J Biol Chem. 1985; 260(6):3289-94. View

13.

Das D, Engelman R, Rousou J, Breyer R, Otani H, Lemeshow S . Role of membrane phospholipids in myocardial injury induced by ischemia and reperfusion. Am J Physiol. 1986; 251(1 Pt 2):H71-9. DOI: 10.1152/ajpheart.1986.251.1.H71. View

14.

Pastorino J, Snyder J, Serroni A, Hoek J, Farber J . Cyclosporin and carnitine prevent the anoxic death of cultured hepatocytes by inhibiting the mitochondrial permeability transition. J Biol Chem. 1993; 268(19):13791-8. View

15.

Nicotera P, Hartzell P, Baldi C, SVENSSON S, Bellomo G, Orrenius S . Cystamine induces toxicity in hepatocytes through the elevation of cytosolic Ca2+ and the stimulation of a nonlysosomal proteolytic system. J Biol Chem. 1986; 261(31):14628-35. View

16.

Floyd R . Role of oxygen free radicals in carcinogenesis and brain ischemia. FASEB J. 1990; 4(9):2587-97. View

17.

Sippel H, Stauffert I, ESTLER C . Protective effect of various calcium antagonists against an experimentally induced calcium overload in isolated hepatocytes. Biochem Pharmacol. 1993; 46(11):1937-44. DOI: 10.1016/0006-2952(93)90634-9. View

18.

Thomas C, Reed D . Effect of extracellular Ca++ omission on isolated hepatocytes. I. Induction of oxidative stress and cell injury. J Pharmacol Exp Ther. 1988; 245(2):493-500. View

19.

Farber J, Kyle M, Coleman J . Mechanisms of cell injury by activated oxygen species. Lab Invest. 1990; 62(6):670-9. View

20.

Landon E, Naukam R, RAMA SASTRY B . Effects of calcium channel blocking agents on calcium and centrilobular necrosis in the liver of rats treated with hepatotoxic agents. Biochem Pharmacol. 1986; 35(4):697-705. DOI: 10.1016/0006-2952(86)90369-2. View