Cobreros A, Sainz L, Lasheras B, Cenarruzabeitia E
Department of Pharmacology, University of Navarra, Pamplona, Spain.
Liver. 1997 Apr;17(2):76-82. doi: 10.1111/j.1600-0676.1997.tb00784.x.
This study examines the effects of three calcium channel blockers (verapamil, nifedipine and diltiazem) on isolated rat hepatocytes exposed to ethanol. In the first part of our study, hepatocytes were incubated with increasing concentrations of ethanol (100, 300, 500, 1000 mM) for varying times. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release were measured to evaluate the cytotoxic effects of ethanol. The concentration of 300 mM and time of incubation of 45 min were chosen for cytoprotection experiments in which calcium channel blockers, at two different concentrations, were added to the medium 30 min prior to the addition of ethanol. ALT, AST and LDH release as well as lipid peroxidation and cellular reduced glutathione (GSH) were measured. Nifedipine and verapamil (25 microM) reduced ALT, AST and LDH activities. The highest dose of diltiazem (50 microM) was more effective than the lowest one (25 microM). Ethanol caused a significant depletion of cellular GSH content as well as a moderate enhancement of lipid peroxidation. While none of the three calcium channel blockers was able to restore the decrease in GSH levels, diltiazem (25 microM) and nifedipine (50 microM) showed the greatest effect, significantly reducing lipid peroxidation.
本研究考察了三种钙通道阻滞剂（维拉帕米、硝苯地平和地尔硫䓬）对暴露于乙醇的离体大鼠肝细胞的影响。在我们研究的第一部分，将肝细胞与浓度不断增加的乙醇（100、300、500、1000 mM）孵育不同时间。测量丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）和乳酸脱氢酶（LDH）的释放，以评估乙醇的细胞毒性作用。在细胞保护实验中选择300 mM的浓度和45分钟的孵育时间，在加入乙醇前30分钟向培养基中添加两种不同浓度的钙通道阻滞剂。测量ALT、AST和LDH的释放以及脂质过氧化和细胞内还原型谷胱甘肽（GSH）。硝苯地平和维拉帕米（25 μM）降低了ALT、AST和LDH的活性。地尔硫䓬的最高剂量（50 μM）比最低剂量（25 μM）更有效。乙醇导致细胞GSH含量显著减少以及脂质过氧化适度增强。虽然三种钙通道阻滞剂均未能恢复GSH水平的降低，但地尔硫䓬（25 μM）和硝苯地平（50 μM）显示出最大效果，显著降低了脂质过氧化。
