Table 1.
Effect of antioxidant, ROS scavengers, ferric chelator, MPT pore sealing agents, lysosomotropic agents, and P450 reductase inhibitor on DTIC and Compound III -induced hepatocyte cytotoxicity and ROS formation
| Addition | Cytotoxicity % (3h) | ROS (30min) |
|---|---|---|
| None | 20± 2 | 79 ± 4 |
| Dacarbazine (56 μM ) | 76 ± 4(1) | 230 ± 4(1) |
| +Catalase (200 U/mL) | 46 ± 2(2) | 116 ± 5(2) |
| +Superoxide dismutase (100 U/mL) | 45 ± 3(2) | 122 ± 2(2) |
| +BHT (50 μM) | 42 ± 3(2) | 118 ± 4(2) |
| +Mannitol (50 mM) | 48 ± 3(2) | 136 ± 3(2) |
| +Dimethyl sulfoxide (150 μM) | 44 ± 3(2) | 121 ± 2(2) |
| +Phenylimidazole (300 μM) | 52 ± 3(2) | 161 ± 3(2) |
| +Diphenyliodoniumchloride (50 μM) | 48 ± 5(2) | 166 ± 3(2) |
| +Methylamine (30 mM) | 36 ± 4(2) | 117 ± 3(2) |
| +Chloroquine (100 μM) | 40 ± 3(2) | 128 ± 2(2) |
| +Desferoxamine (200 μM) | 36 ± 2(2) | 121 ± 3(2) |
| +Cyclosporine (2 μM) | 34 ± 3(2) | 138 ± 3(2) |
| +Carnitine (2 mM) | 37 ± 4(2) | 152 ± 3(2) |
| Compound III (33 μM) | 73 ± 2(1) | 256 ± 5(1) |
| +Catalase (200 U/mL) | 38 ± 2(3) | 126 ± 3(3) |
| +Superoxide dismutase (100 U/mL) | 41 ± 4(3) | 132 ± 2(3) |
| +BHT(50 μM) | 37 ± 4(3) | 128 ± 2(3) |
| +Mannitol (50 mM) | 38 ± 4(3) | 141 ± 3(3) |
| +Dimethyl sulfoxide (150 μM) | 36 ± 3(3) | 145 ± 2(3) |
| +Phenylimidazole (300 μM) | 48 ± 5(3) | 162 ± 3(3) |
| +Diphenyliodoniumchloride (50 μM) | 48 ± 5(3) | 167 ± 4(3) |
| +Methylamine (30 mM) | 31 ± 2(3) | 141 ± 2(3) |
| +Chloroquine (100 μM) | 46 ± 3(3) | 155 ± 3(3) |
| +Desferoxamine (200 μM) | 35 ± 3(3) | 136 ± 3(3) |
| +Cyclosporine (2 μM) | 28 ± 2(3) | 141 ± 2(3) |
| +Carnitine (2 mM) | 31 ± 3(3) | 161 ± 3(3) |
Hepatocytes (106 cells/mL) were incubated in Krebs-Henseleit buffer pH 7.4 at 37 ˚C for 3 h following the addition of DTIC and Compound III. Cytotoxicity was determined as the percentage of cells that take up trypan blue (19, 33). DCF formation was expressed as fluorescent intensity units (34). Values are expressed as means of three separate experiments (SD).
(1) Significant difference in comparison with control hepatocytes (p < 0.05).
(2) Significant difference in comparison with DTIC treated hepatocytes (p < 0.05).
(3) Significant difference in comparison with compound III treated hepatocytes (p < 0.05).