Table 1.
Hepatic metabolite concentrations
| Experimental group | WT-CON | KO-CON | WT-EtOH | KO-EtOH |
|---|---|---|---|---|
| Amino acids and derivatives (nmol/g liver)1 | ||||
| Arginine | 87 ± 22.7 | 76 ± 16.8 | 98 ± 10.2 | 73 ± 13.9 |
| Aspartate | 471 ± 108.4 | 564 ± 162.9 | 509 ± 62.6 | 686 ± 260.9 |
| Betaine | 195 ± 36 | 102 ± 35a | 111 ± 57 | 139 ± 67 |
| Carnitine | 265 ± 24.2 | 240 ± 43.1 | 230 ± 44.0 | 277 ± 73.1 |
| Choline | 3748 ± 1896 | 3831 ± 1240 | 8944 ± 2726b | 8013 ± 2617b |
| Citruline | 11 ± 1.9 | 15 ± 2.9 | 28 ± 21.5 | 31 ± 35.6 |
| Creatine | 689 ± 101.8 | 603 ± 72.6 | 665 ± 158.6 | 816 ± 399.6 |
| Creatinine | 17 ± 1.0 | 17 ± 2.0 | 19 ± 3.5 | 19 ± 7.2 |
| Glutamine | 4082 ± 472.0 | 4335 ± 1016.6 | 4295 ± 252.6 | 4848 ± 1412.1 |
| Histidine | 381 ± 25.0 | 419 ± 75.0 | 410 ± 46.2 | 457 ± 54.4 |
| Lysine | 611 ± 71.1 | 614 ± 103.6 | 672 ± 240.5 | 644 ± 174.7 |
| Methionine | 51 ± 10.2 | 51 ± 6.1 | 69 ± 19.5 | 69 ± 9.5 |
| N-acetylglutamine | 805 ± 167.8 | 873 ± 180.4 | 733 ± 268.2 | 937 ± 512.2 |
| Nε-acetyllysine | 2 ± 0.4 | 3 ± 0.4 | 3 ± 1.0 | 4 ± 1.9 |
| N-acetylcysteine | 1.0 ± 0.3 | 0.7 ± 0.1 | 1.6 ± 0.7 | 0.7 ± 0.1 |
| Ornithine | 279 ± 28.8 | 375 ± 79.7 | 359 ± 124.9 | 449 ± 131.7 |
| Proline | 160 ± 22.0 | 202 ± 38.5 | 158 ± 61.4 | 239 ± 59.1 |
| Taurine | 11284 ± 980 | 14808 ± 1260a | 10609 ± 1546 | 9967 ± 4418b |
| Glycolysis and TCA cycle (nmol/g liver)1 | ||||
| ADP | 369 ± 33 | 486 ± 109 | 514 ± 56 | 606 ± 144 |
| ATP | 205 ± 13 | 226 ± 54 | 215 ± 45 | 210 ± 50 |
| Isocitrate | 306 ± 67.7 | 273 ± 33.8 | 311 ± 60.5 | 281 ± 111.9 |
| Lactate | 57 ± 11 | 69 ± 7 | 36 ± 4b | 41 ± 7b |
| Malate | 4948 ± 1062.8 | 4725 ± 1348.7 | 4354 ± 793.3 | 5197 ± 1211.8 |
| Succinic acid | 408 ± 83.5 | 617 ± 242.6 | 350 ± 90.8 | 629 ± 333.4 |
| Nucleic acid metabolism (nmol/g liver)1 | ||||
| Adenosine | 197 ± 29 | 224 ± 68 | 243 ± 46 | 344 ± 86b |
| AMP | 34 ± 12.7 | 34 ± 12.9 | 42 ± 18.1 | 47 ± 20.5 |
| CMP | 72 ± 10.2 | 98 ± 27.7 | 97 ± 22.1 | 90 ± 31.2 |
| Cytidine | 2 ± 0.5 | 3 ± 0.7 | 2 ± 0.3 | 3 ± 0.6 |
| deoxycytidine | 0.4 ± 0.1 | 0.4 ± 0.1 | 0.4 ± 0.1 | 0.5 ± 0.1 |
| deoxyuridine | 7117 ± 4393.5 | 8117 ± 3693.1 | 6490 ± 4275.5 | 5793 ± 2604.3 |
| GMP | 898 ± 34 | 900 ± 216 | 1166 ± 125 | 1464 ± 513b |
| Hypoxanthine | 689 ± 101.8 | 603 ± 72.6 | 665 ± 158.6 | 816 ± 399.6 |
| IMP | 1125 ± 74.8 | 1049 ± 395.1 | 1055 ± 229.8 | 1245 ± 538.5 |
| TMP | 7 ± 1.1 | 6 ± 0.6 | 7 ± 0.7 | 7 ± 1.4 |
| UMP | 11085 ± 128 | 15801 ± 4001 | 13013 ± 2758 | 17652 ± 5690 |
| Uracil | 3 ± 0.9 | 2 ± 0.5 | 2 ± 1.1 | 3 ± 2.8 |
| UTP | 54 ± 7.5 | 52 ± 5.7 | 55 ± 4.4 | 53 ± 8.6 |
| Xanthine | 203 ± 18.6 | 150 ± 38.3 | 157 ± 31.4 | 157 ± 77.8 |
| Xanthosine | 181 ± 44 | 182 ± 66 | 88 ± 21b | 104 ± 44b |
| Nucleotide cofactors (nmol/mg proteins)2 | ||||
| NADPH | 1.37 ± 0.42 | 1.10 ± 0.29 | 1.39 ± 0.39 | 1.24 ± 0.27 |
| NADP | 0.33 ± 0.07 | 0.30 ± 0.07 | 0.38 ± 0.11 | 0.40 ± 0.1 |
| NADPH/NADP ratio | 4.2 ± 0.7 | 3.8 ± 1.3 | 3.8 ± 1.2 | 3.2 ± 0.9 |
1
Selected metabolites in the liver extract were quantitated by UPLC-MS/MS. The concentration of each metabolite was determined from respective calibration curve and normalized by liver weight. Results are reported in nmole per gram liver weight.
2
Concentrations of NADH and NADPH were measured using a biochemical assay kit (Abcam, Cambridge, MA); results are reported in nmole per milligram total proteins.
Data are presented as mean ± S.D. (N = 4/group). Group differences were analyzed using Graphpad Prism software by two-way ANOVA with post-hoc Holm-Sidak's multiple comparisons test.
a
P < 0.05, when compared with diet-matched WT mice;
b
P < 0.05, when compared with pair-fed mice of the same genotype.