Table 3. Metabolite profiling and redox state in freeze-clamped liver following perfusion with or without GKA.

Metabolites were determined in extracts of freeze-clamped liver after perfusion without (Control) or with GKA [(+) GKA] as indicated in the Experimental section. The redox state is represented by the free cytosolic NAD⁺/NADH and NADP⁺/NADPH ratios calculated from concentrations of pyruvate, lactate or malate. The mitochondrial NAD⁺/NADH ratio was calculated from concentrations of β-OH-butyrate and acetoacetate as indicated [32]. Results are means ± S.D. of three livers per group. P values were determined by Student’s t test using Prism 5 software.

(a) Metabolite level
	Control (μmol per g of wet mass)	(+) GKA (μmol per g of wet mass)	P value
Lactate	1.51 ± 0.06	1.81 ± 0.15	0.02
Pyruvate	0.28 ± 0.03	0.42 ± 0.12	0.06
Malate	0.18 ± 0.05	0.26 ± 0.03	N.S.
Citrate	0.25 ± 0.04	0.29 ± 0.04	N.S.
Acetoacetate	0.35 ± 0.03	0.37 ± 0.13	N.S
β-OH-butyrate	0.23 ± 0.04	0.25 ± 0.04	N.S.
NAD	0.18 ± 0.06	0.26 ± 0.03	N.S.
NADH	0.083 ± 0.05	0.081 ± 0.02	N.S.
	Control (nmol per g of wet mass)	(+) GKA (nmol per g of wet mass)	P value
G6P	33.7 ± 6.2	103.2 ± 15.9	0.007
GA3P	3.2 ± 1.3	7.2 ± 0.9	0.014
Fructose 1,6-bisphosphate	65.7 ± 21	75.4 ± 5.8	N.S.
	Control (mg per g of wet mass)	(+) GKA (mg per g of wet mass)	P value
Triglycerides	0.26 ± 0.03	0.38 ± 0.16	N.S.

(b) Redox state
	Control	(+) GKA	P value
Cytoplasmic NAD+/NADH	1684 ± 126	2170 ± 819	N.S.
Cytoplasmic NADP+/NADPH	0.055 ± 0.01	0.066 ± 0.02	N.S.
Mitochondrial NAD+/NADH	32 ± 7.2	31 ± 12.5	N.S.

N.S., not significant.