Table 4.
Blockade ofDTIC and Compound III induced Caspase-3 activation by antioxidants, ROS scavengers, CYP2E1 inhibitor, p450 reductase inhibitor and mitochondrial MPT pore sealing agents
| Addition |
Caspase-3 Activity
2 h |
|---|---|
| None | 297.65 ± 5 |
| Dacarbazine (56 μM ) | 659.52 ± 7(1) |
| +Catalase (200 U/mL) | 258.74 ± 5(2) |
| +BHT (50 μM) | 260.52 ± 3(2) |
| +Mannitol (50 mM) | 285.46 ± 4(2) |
| +Dimethyl sulfoxide (150 μM) | 281.33 ± 5(2) |
| +Phenylimidazole (300 μM) | 223.39 ± 3(2) |
| +Diphenyliodoniumchloride (50 μM) | 244.65 ± 4(2) |
| +Cyclosporine (2 μM) | 116.32 ± 2(2) |
| +Carnitine (2 mM) | 124.38 ± 3(2) |
| Compound III(33 μM) | 634.78 ± 5(1) |
| +Catalase (200 U/mL) | 138.73 ± 4(3) |
| +BHT (50 μM) | 149.13 ± 3(3) |
| +Mannitol (50 mM) | 281.22 ± 3(3) |
| +Dimethyl sulfoxide (150 μM) | 251.18 ± 2(3) |
| +Phenylimidazole (300 μM) | 173.89 ± 3(3) |
| +Diphenyliodoniumchloride (50 μM) | 188.36 ± 3(3) |
| +Cyclosporine (2 μM) | 96.18 ± 2(3) |
| +Carnitine (2 mM) | 99.63 ± 2(3) |
Hepatocytes (106 cells/mL) were incubated in Krebs-Henseleit buffer pH 7.4 at 37 ˚C for 30 min. The activating of caspase-3 (μM pNA/min/mL)) was determined based on hydrolysis of pNA labeled substrate (36). 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).