ABSTRACT
The effect of the hepatotoxic substance diamidinothionaphthene (98/202) on cytosolic, mitochondrial and extra-mitochondrial calcium distribution was measured in isolated rat hepatocytes. The drastic disturbance of the intracellular calcium homeostasis caused by this substance (increase of the cytosolic and mitochondrial calcium contents and depletion of extra-mitochondrial calcium stores, which at last lead to cell death) gave rise to an investigation of the possible cytoprotective effect of calcium antagonists of various chemical classes: verapamil, diltiazem, and nifedipine on isolated hepatocytes. Our results show that all three calcium antagonists prevented cell death caused by 98/202. The 98/202-induced increase of cytosolic and mitochondrial calcium content was inhibited by all three calcium antagonists. However, only verapamil was able to inhibit the depletion of extra-mitochondrial calcium stores. Since 98/202-induced cell death occurs only in the presence of extracellular calcium, it is concluded that calcium antagonists are also able to inhibit the influx of extracellular calcium in liver cells, which leads to a calcium overload of the cytosol and mitochondria. The various ways of interfering with the calcium homeostasis of liver cells qualifies the hepatotoxic substance 98/202 as a suitable in vitro hepatotoxicity model for testing the hepatoprotective effect of different calcium antagonists.
Subject(s)
Amidines/antagonists & inhibitors , Calcium Channel Blockers/pharmacology , Liver/drug effects , Animals , Calcium/metabolism , Cell Survival , Cells, Cultured/drug effects , Cytosol/drug effects , Liver/metabolism , Male , Mitochondria, Liver/drug effects , Phosphorylase a/metabolism , Rats , Rats, Wistar , Ruthenium RedABSTRACT
Pentamidine, DAPI and some related compounds (DAI, 6-Br-AI, DPTN, DIPI, 3-Am-DAI, DiaPBF) were investigated in 2 different screening test systems for their potential mutagenic and cytotoxic effects, in the light of their binding to DNA. In the Ames test using Salmonella typhimurium strains TA98 and TA100 with and without metabolic activation no mutagenic effects could be observed. All diamidines tested, except DAI, were toxic at concentrations of 0.5 and 1.0 mumole/plate. In the sister-chromatid exchange (SCE) assay with human peripheral lymphocytes all compounds tested were growth-retarding particularly in the G0 phase. A significant induction of SCEs could only be seen after treatment with the monoamidino compound 6-Br-AI at a concentration of 100 mumole/l. It is concluded from the data obtained that pentamidine and related diamidines in the 2 assays tested show no mutagenic or genotoxic effects, in spite of their tight binding to DNA.
