A (1)H NMR-Based Metabonomic Investigation of Time-Related Metabolic Trajectories of the Plasma, Urine and Liver Extracts of Hyperlipidemic Hamsters - PubMed (original) (raw)

Figure 1

Figure 1. Ultrasonographic imaging of liver tissue from control (left) and HFHC hamsters (right) at week 42.

Figure 2

Figure 2. Oil Red O staining of liver tissue from control (left) and HFHC hamsters (right) at week 42 revealed the accumulation of fat in the latter group.

Figure 3

Figure 3. Representative hematoxylin and eosin (H&E) staining of liver sections from control (left) and HFHC hamsters (right) at week 42.

Lipid droplets were obviously accumulated in HFHC hamsters, but not control hamsters.

Figure 4

Figure 4. 500 MHz 1H NMR spectra of plasma samples.

(a) 1H-NMR spectra of plasma samples after filtration. (b) 1H-NMR CPMG spectra of plasma samples. (c) 1H-NMR BPP-LED spectra of plasma samples. Abbreviations: 2-HB, 2-hydroxybutyrate; Isobu, isobutyrate; Ile, isoleucine; Leu, leucine; Val, valine; Eth, ethanol; 3-HB, 3-hydroxybutyrate; Lac, lactate; Ala, alanine; Ace, acetate; AcAc, acetoacetate; Pro, proline; Pyr, pyruvate; Glu, glutamine; Cir, citrate; Met, methionine; Sar, sarcosine; Lys, lysine; Cre, creatine; Cho, choline; Tyr, tyrosine; Phe, phenylalanine; Ino, inosine; For, formate; VLDL, very low-density lipoprotein; LDL, low-density lipoprotein; HDL, high-density lipoprotein; PtdCho, phosphatidylcholine; and N-Ac, N-acetyl glycoproteins.

Figure 5

Figure 5. 500 MHz 1H NMR spectra of urine samples.

1, DSS; 2, 2-Hydroxyisovalerate; 3, 2-Hydroxybutyrate; 4, 2-Hydroxyvalerate; 5, 2-Oxoglutarate; 6, Isoleucine; 7, Valine;8, Isobutyrate; 9, Methylsuccinate; 10, Ethanol; 11, 3-Hydroxybutyrate; 12, Fucose; 13, 3-Hydroxyisovalerate; 14, Lactate; 15, 3-Hydroxy-3-methylglutarate; 16, 2-Hydroxyisobutyrate; 17, Alanine; 18, Acetate; 19, N-Acetylglutamate; 20, N-Acetylglycine; 21, Glutamine; 22, Acetone; 23, Acetoacetate; 24, Glycerol; 25, Citrate; 26, Methylamine; 27, Dimethylamine; 28, N,N-Dimethylglycine; 29, Creatinine; 30, cis-Aconitate; 31, O-Acetylcarnitine; 32, Choline; 33, O-Phosphocholine; 34, Carnitine; 35, Betaine; 36, Trimethylamine N-oxide; 37, Taurine; 38, Methanol; 39, 4-Hydroxyphenylactate; 40, 3-Methylxanthine; 41, N-Phenylacetylglycine; 42, Hippurate; 43, Tartrate; 44, Xylose; 45, Allantoin; 46, Urea; 47, Cytosine; 48, ADP; 49, Urocanate; 50, Fumarate; 51, trans-Aconitate; 52, Tyrosine; 53, 3-Indoxylsulfate; 54, 3-Phenyllactate; 55, Nicotinamide N-oxide; 56, Hypoxanthine; 57, Nicotinurate; 58, Inosine; 59, Formate.

Figure 6

Figure 6. 500 MHz 1H NMR spectra of aqueous liver extract.

1, 2-Hydroxyisovalerate; 2, 2-Hydroxybutyrate; 3, Isoleucine; 4, Leucine; 5, 2-Aminobutyrate; 6, Valine; 7, Methylsuccinate; 8, Ethanol; 9, 3-Hydroxybutyrate; 10, Lactate; 11, Alanine; 12, Cadaverine; 13, Ornithine; 14, Acetate; 15, Glutamate; 16, Glutamine; 17, Glutathione; 18, Succinate; 19, Creatine; 20, Aspartate; 21, Malate; 22, Dimethylamine; 23, Choline; 24, O-Phosphocholine; 25, Carnitine; 26, Glucose; 27, Betaine; 28, Methanol; 29, Glycine; 30, Glycerol; 31, Ethylene glycol; 32, UDP-glucose; 33, UDP-glucuronate; 34, UDP-galactose; 35, Uridine; 36, NAD+; 37, Fumarate; 38, Tyrosine; 39, 3-Indoxylsulfate; 40, 3-Phenyllactate; 41, Nicotinurate; 42, NADP+; 43, Adenosine; 44, Inosine; 45, Formate; 46, ADP/AMP.

Figure 7

Figure 7. 500 MHz 1H NMR spectra of lipophilic liver extract.

1, Total Cholesterol (C18 H 3); 2, Total Cholesterol(C26 H 3, C27 H 3 ); 3, Fatty acid residues (ω-CH 3); 4, Total Cholesterol (C21 H 3); 5, Fatty acid residues (ω-CH 3 of DHA+EPA+linolenic); 6, Free Cholesterol (C19 H 3); 7, Esterified Cholesterol (C19 H 3); 8, Fatty acid residues ((CH 2-)n); 9, Fatty acid residues (COCH2-CH 2); 10, Fatty acid residues (−CH 2 of ARA+EPA); 11, Fatty acid residues (CH 2-CH = ); 12, Fatty acid residues (γ CH 2 of ARA+EPA); 13, Monoglycerides(FA, RH -CH2-CO-O-C2); 14, Fatty acid residues (−CO-CH 2); 15, Fatty acid residues (α and β CH 2 of DHA); 16, Fatty acid residues (−CH = CH-CH 2-CH = CH-of linoleic acid); 17, FA, PUFA (CH = CH-CH 2-(CH = CH-CH2)n, n>1); 18, Phosphatidylethanolamine (−CH 2-NH2); 19, Sphingomyelin (−CH2-N-(CH 3)3); 20, Phosphatidylcholine (−CH2-N-(CH 3)3); 21, Cholesterol (C3 H); 22, Total phospholipids (Glycerol (C3 H2)); 23, Triglycerides (C1 H and C3 H of glycerol); 24, Triglycerides (C1 H and C3 H of glycerol); 25, Triglycerides(C2 H of glycerol); 26, Fatty acid residues (−CH = CH-); 27, Cholesterol (C6 H). ARA, Arachidonic acid; EPA, Eicosapentaenoic acid; DHA, Docosahexaenoic acid.

Figure 8

Figure 8. Representative quantifications and corresponding spectral regions of some plasma metabolites together with the mathematical fit with Chenomx.

Assignments: 2-hydroxybutyrate (yellow); isoleucine (red); leucine (green); valine (blue); isobutyrate (pink).

Figure 9

Figure 9. Trajectory derived from PCA of 1H-NMR spectra of hamster plasma from control groups at different time points (▴, week 0; ▪, week 3; ♦, week 9; •, week 15; *, week 26; ○, week 35; □, week 42).

(a) Trajectory of 1H-CPMG-NMR spectra at different time points (two PCs, R2X = 0.896; Q2 = 0.391). (b) Trajectory of 1H- LEDBP-NMR spectra at different time points (two PCs, R2X = 0.968; Q2 = 0.835).

Figure 10

Figure 10. Trajectory derived from PCA of 1H-CPMG-NMR spectra of hamster plasma normalized on the sum of the spectrum mapping the time-related trajectory of metabotypes at weeks 0, 3, 6, 9, 15, 21, 26, 35 and 42.

Figure 11

Figure 11. Trajectory derived from PCA of 1H-LEDBP-NMR spectra of hamster plasma normalized on the sum of the spectrum mapping the time-related trajectory of metabotypes at weeks 0, 3, 6, 9, 15, 21, 26, 35 and 42.

Figure 12

Figure 12. PCA score plots of plasma based on targeted profiling of 40 measured metabolites (▴, week 0; ♦, week 4; ▪, week 24; and •, week 42. R2X = 0.355, Q2 = 0.346).

Figure 13

Figure 13. OPLS-DA scores (upper panel) and correlation coefficient plots (lower panel) derived from NMR data for hamster plasma samples at week 24 (A) and week 42 (B) (▴, control; ♦, HFHC).

Positive bars (± SEM) of correlation coefficient plots denote metabolites significantly higher in control group, whereas negative bars (± SEM) denote metabolites significantly increased in the HFHC group.

Figure 14

Figure 14. Trajectory derived from PCA of targeted profiling of 80 measured urine metabolites (normalized to the total concentration of all measured metabolites) revealed metabolic changes associated with dietary treatment.

Figure 15

Figure 15. PCA score plots of 1H-NMR spectra of hamster urine from control groups at different time points(▴, week 0; ▪, week 6; ♦, week 15; •, week 26; *, week 35; □, week 42).

Figure 16

Figure 16. OPLS-DA scores (upper) and correlation coefficient plots (lower) derived from NMR data for hamster urine samples at week 15 (A) and week 42 (B) (▴, control; ♦, HFHC).

Positive bars (± SEM) of correlation coefficient plots denote metabolites significantly higher in control group, whereas negative bars (± SEM) denote illustrate metabolites.

Figure 17

Figure 17. Temporal changes in individual metabolite levels in the urine samples of HFHC animals at weeks 3, 6, 9, 15, 21, 26, 35 and 42.

The Y axis values were calculated based on the measured metabolite concentrations in the untreated control group with the formula Cm/Cc, where Cm represents the respective metabolite concentration in the HFHC group and Cc represents that in the control group. The error bars represent the standard deviation of the mean.

Figure 18

Figure 18. Multivariate data analyses of the 1H-NMR spectra of the water-soluble fraction of liver extracts at different time points: ▴, week 0; ♦, week 4; ▪, week 24; and •, week 42. R2X = 0.582; Q2 = 0.494.

Figure 19

Figure 19. PCA plots derived from 1H -NMR spectra of the organic phase of liver extracts at different time points: ▴, week 0; ♦, week 4; ▪, week 24; and •, week 42; R2X = 0.753; Q2 = 0.703.

Figure 20

Figure 20. Schematic diagram of the hamster metabolic pathways and the relative levels of the major compounds within these pathways at various times in the development of hyperlipidaemia.

1, 2, and 3 stand for plasma samples taken at weeks 4, 21 and 35, respectively, while a, b and c stand for urine samples taken at weeks 6, 21 and 42, respectively.