Table 3.

Mitochondrial membrane potential changes during DTIC and Compound III induced hepatocyte injury by antioxidants, ROS scavengers, CYP2E1 inhibitor, P450 reductase inhibitor and mitochondrial MPT pore sealing agents

ΔΨm%Incubation Time			Addition
min 30	min 15	min 2
4 ± 2	3 ± 1	2 ± 1		None
77 ± 3(1)	56 ± 2(1)	42 ± 3(1)		Dacarbazine (56 μM )
16 ± 2(2)	10 ± 3(2)	6 ± 2(2)		+Catalase (200 U/mL)
20 ± 3(2)	14 ± 2(2)	6 ± 3(2)		+BHT(50 μM)
21 ± 2(2)	16 ± 2(2)	9 ± 3(2)		+Mannitol (50 mM)
18 ± 2(2)	14 ± 3(2)	6 ± 2(2)		+Dimethyl sulfoxide (150 μM)
15 ± 1(2)	9 ± 3(2)	6 ± 3(2)		+Phenylimidazole (300 μM)
19 ± 2(2)	12 ± 1(2)	8 ± 2(2)		+Diphenyliodoniumchloride (50 μM)
16 ± 2(2)	10 ± 1(2)	8 ± 3(2)		+Cyclosporine (2 μM)
19 ± 2(2)	12 ± 2(2)	8 ± 2(2)		+Carnitine (2 mM)
90 ± 4(1)	66 ± 6(1)	55 ± 1(1)		Compound III(33 μM)
18 ± 2(3)	12 ± 2(3)	7 ± 2(3)		+Catalase (200 U/mL)
18 ± 3(3)	15 ± 3(3)	10 ± 3(3)		+BHT(50 μM)
22 ± 2(3)	17 ± 2(3)	9 ± 2(3)		+Mannitol (50 mM)
18 ± 2(3)	15 ± 2(3)	8 ± 3(3)		+Dimethyl sulfoxide (150 μM)
22 ± 2(3)	16 ± 3(3)	10 ± 1(3)		+Phenylimidazole (300 μM)
26 ± 3(3)	18 ± 2(3)	11 ± 1(3)		+Diphenyliodoniumchloride (50 μM)
16 ± 2(3)	11 ± 3(3)	5 ± 2(3)		+Cyclosporine (2 μM)
20 ± 2(3)	14 ± 2(3)	10 ± 1(3)		+Carnitine (2 mM)

Hepatocytes (106 cells/mL) were incubated in Krebs-Henseleit buffer pH 7.4 at 37 ˚C for 30 min. Mitochondrial membrane potential was determined as the difference in mitochondrial uptake of the rhodamine 123 between control and treated cells. Our data were shown as the percentage of mitochondrial membrane potential collapse (%ΔΨm) in all treated (test) hepatocyte groups (22, 35).

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).