Evaluation of calcium channel blockers as potential hepatoprotective agents in oxidative stress injury of perfused hepatocytes.
The aim of this study was to investigate the effects of calcium channel blockers on tertbutyl hydroperoxide (TBH) induced liver injury using isolated perfused rat hepatocytes. Rat hepatocytes were immobilized in agarose threads and perfused with Williams E medium. Hepatocyte injury was induced by the addition of tertbutyl hydroperoxide (1 mM) to the perfusion medium 30 min after the addition of either verapamil or diltiazim. Hepatocyte injury was observed by monitoring the functional and metabolic competence of hepatocytes or by ultrastructural morphological examination of hepatocytes. Verapamil (0.5 mM) reduced lactate dehydrogenase leakage in TBH-injured hepatocytes as compared to the controls (154+/-11% vs. 247+/-30%). Lipid peroxides production was reduced after verapamil pretreatment as compared to the controls and oxygen consumption was increased by pretreatment of hepatocytes with verapamil. Verapamil pretreatment increased the protein synthesis activity at both levels of granular endoplasmic reticulum and free polysomes in cytoplasm and decreased ATPase activity. Diltiazem was qualitatively effective as verapamil. It is concluded that in hepatocyte oxidative injury, calcium channel blockers exhibited hepatoprotective properties. The hepatoprotective effect of calcium channel blockers was accompanied by a decrease in ATPase activity, which may implicate a normalization of Ca2+i after TBH intoxication.
Farghali
H
H
Institute of Pharmacology, First Faculty of Medicine, Charles University, Prague, Czech Republic. hfarg@lf1.cuni.cz
Kmonícková
E
E
Lotková
H
H
Martínek
J
J
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D056486:Chemical and Drug Induced Liver Injury; D004110:Diltiazem / Q000494:pharmacology; D022781:Hepatocytes / Q000187:drug effects* / Q000648:ultrastructure; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008054:Lipid Peroxides / Q000037:antagonists & inhibitors; D008107:Liver Diseases / Q000517:prevention & control*; D008297:Male; D018384:Oxidative Stress; D010101:Oxygen Consumption / Q000187:drug effects; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D014700:Verapamil / Q000494:pharmacology; D020122:tert-Butylhydroperoxide
D002121:Calcium Channel Blockers; D008054:Lipid Peroxides; D020122:tert-Butylhydroperoxide; D014700:Verapamil; D007770:L-Lactate Dehydrogenase; D000251:Adenosine Triphosphatases; D004110:Diltiazem