Table 2.
Preventing DTIC and Compound III induced hepatocyte lysosomal membrane damage by antioxidants, ROS scavengers, ferric chelator, CYP2E1 inhibitor, P450 reductase inhibitor, lysosomotropic agents
| Addition |
% Acridine orange redistribution
Incubation Time |
||
|---|---|---|---|
| 2 min | 15 min | 30 min | |
| None | 2 ± 1 | 4 ± 2 | 4 ± 3 |
| Dacarbazine (56 μM ) | 183 ± 5(1) | 237 ± 5(1) | 250 ± 4(1) |
| +Catalase (200 U/mL) | 11 ± 1(2) | 14 ± 2(2) | 18 ± 2(2) |
| +SOD (100 U/mL) | 10 ± 2(2) | 16 ± 2(2) | 20 ± 2(2) |
| +BHT (50 μM) | 14 ± 1(2) | 19 ± 2(2) | 25 ± 3(2) |
| +Mannitol (50 mM) | 8 ± 2(2) | 11 ± 1(2) | 13 ± 1(2) |
| +Dimethyl sulfoxide (150 μM) | 8 ± 3(2) | 10 ± 1(2) | 12 ± 1(2) |
| +Phenylimidazole (300 μM) | 16 ± 2(2) | 22 ± 1(2) | 30 ± 3(2) |
| +Diphenyliodoniumchloride (50 μM) | 18 ± 3(2) | 26 ± 3(2) | 33 ± 3(2) |
| +Methylamine (30 mM) | 8 ± 2(2) | 11 ± 1(2) | 14 ± 1(2) |
| +Chloroquine (100 μM) | 12 ± 1(2) | 15 ± 2(2) | 20 ± 2(2) |
| +Desferoxamine (200 μM) | 8 ± 2(2) | 10 ± 2(2) | 11 ± 1(2) |
| Compound III (33 μM) | 194 ± 5(1) | 240 ± 5(1) | 264 ± 5(1) |
| +Catalase (200 U/mL) | 12 ± 1(3) | 16 ± 2(3) | 18 ± 2(3) |
| +SOD (100 U/mL) | 10 ± 2(3) | 15 ± 2(3) | 21 ± 2(3) |
| +BHT (50 μM) | 16 ± 2(3) | 20 ± 3(3) | 24 ± 3(3) |
| +Mannitol (50 mM) | 12 ± 1(3) | 16 ± 1(3) | 19 ± 2(3) |
| +Dimethyl sulfoxide (150 μM) | 14 ± 2(3) | 18 ± 2(3) | 21 ± 2(3) |
| +Phenylimidazole (300 μM) | 15 ± 2(3) | 17 ± 2(3) | 24 ± 2(3) |
| +Diphenyliodoniumchloride (50 μM) | 18 ± 3(3) | 22 ± 2(3) | 28 ± 3(3) |
| +Methylamine (30 mM) | 12 ± 1(3) | 14 ± 1(3) | 17 ± 2(3) |
| +Chloroquine (100 μM) | 16 ± 2(3) | 18 ± 2(3) | 25 ± 2(3) |
| +Desferoxamine (200 μM) | 10 ± 1(3) | 12 ± 1(3) | 16 ± 2(3) |
Hepatocytes (106 cells/mL) were incubated in Krebs-Henseleit buffer pH 7.4 at 37 ˚C for 30 min. Lysosomal membrane damage was determined as intensity unit of diffuse cytosolic green fluorescence induced by acridine orange following the release from lysosome (16).
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).