Plasma Metabolomics Study on the Impact of Different CRF Levels on MetS Risk Factors
<p>Flowchart of different metabolic syndrome MetS and cardiorespiratory fitness CRF groupings. Participants were categorized into the low-risk group and the high-risk group (HM) if they had ≥3 MetS-related risk factors; CRF was categorized into low CRF level (LC), medium CRF level (MC), and high CRF level (HC) using the 3-quartile method; participants were classified into four groups based on the CRF level and the number of MetS-related risk factors: low CRF and low MS risk factor group (LCLM), low CRF and high MS risk factor group (LCHM), high CRF and low MS risk factor group (HCLM), and high CRF and high MS risk factor group (HCHM).</p> "> Figure 2
<p>(<b>a</b>) Total signal intensity in the amine/phenol-based channel sub-sample analysis; (<b>b</b>) Background peak mass distribution in the amine/phenol-based channel sub-sample analysis; (<b>c</b>) Retention time assay results; (<b>d</b>) Distribution of identified metabolites in different tiers of the database.</p> "> Figure 3
<p>PLS-DA Score Plot: (<b>a</b>) LC vs. MC vs. HC group; (<b>b</b>) LM vs. HM group (<b>c</b>) LCLM vs. HCLM; (<b>d</b>) LCHM vs. HCHM; (<b>e</b>) LCLM vs. LCHM; (<b>f</b>) HCLM vs. HCHM; (<b>g</b>) LCLM vs. HCHM; (<b>h</b>) LCHM vs. HCLM.</p> "> Figure 4
<p>The OPLS-DA score plots (<b>A</b>–<b>J</b>) depict the scores for each group, while the response ranking test plots (<b>a</b>–<b>j</b>) display the results of the response ranking test. In the ranking test, the horizontal axis represents the correlation between the <span class="html-italic">Y</span> values of the random grouping and the <span class="html-italic">Y</span> values of the original grouping, while the vertical axis represents the R<sup>2</sup> and Q<sup>2</sup> scores.</p> "> Figure 5
<p>Differential metabolite volcano plot (<b>a</b>) HC vs. LC, (<b>b</b>) HM vs. LM, (<b>c</b>) HCLM vs. LCLM, (<b>d</b>) HCHM vs. LCHM, (<b>e</b>) LCHM vs. LCLM, (<b>f</b>) HCHM vs. LCLM. The horizontal axis represents the fold change in metabolite expression across different subgroups [log2(FoldChange)], while the vertical axis indicates the significance level of differences [−log10 (<span class="html-italic">p</span>-value)]. Each point on the plot represents a metabolite, with red indicating a significant increase, blue indicating a significant decrease, and black indicating no significant difference. Plasma metabolites were visualized on a volcano plot based on their fold change (FC) values, <span class="html-italic">p</span>-values, and <span class="html-italic">q</span>-values. VIP value indicates the contribution of each variable to the PLS-DA model. Differential metabolites were identified based on VIP values > 1, fold change (FC) > 1.2 or <0.83, <span class="html-italic">p</span> < 0.05, and <span class="html-italic">q</span> < 0.25.</p> "> Figure 6
<p>Venn diagram analysis of potential biomarkers in different groups.</p> "> Figure 7
<p>Histogram of trends in common differential metabolites by group. “*” indicating differential metabolite FC > 1.2 or <0.83, <span class="html-italic">p</span> < 0.05, and <span class="html-italic">q</span> < 0.25; “**” indicating differential metabolite FC > 1.2 or <0.83, <span class="html-italic">p</span> < 0.01, and <span class="html-italic">q</span> < 0.10; black column: HCHM group vs. LCLM group; grey column: HC group vs. LC group; white column: HM group vs. LM group. (<b>a</b>) 2-oxoglutaric acid; (<b>b</b>) L-arginine; (<b>c</b>) L-serine; (<b>d</b>) cis-aconitic acid; (<b>e</b>) L-glutamine; (<b>f</b>) L-valine.</p> "> Figure 8
<p>Bubble diagram of metabolic pathways of differential metabolites in each group. (<b>a</b>–<b>f</b>) represent the metabolic pathway bubble diagrams for each group, (<b>a</b>) HC vs. LC group; (<b>b</b>) HM vs. LM group; (<b>c</b>) HCLM vs. LCLM group; (<b>d</b>) HCHM vs. LCHM group; (<b>e</b>) LCHM group vs. LCLM group (<b>f</b>) HCHM vs. LCLM group (1): arginine biosynthesis; (2): tricarboxylic acid cycle(TCA); (3): cysteine and methionine metabolism; (4): glycine, serine, and threonine metabolism; (5): arginine and proline metabolism; (6): alanine, aspartic acid, and glutamate metabolism; (7): aminotransferase-tRNA biosynthesis; (8): Butyric acid metabolism; (9): glyoxylate and dicarboxylic acid metabolism; (10): purine metabolism; (11): glutathione metabolism; (12): glycerophospholipid metabolism; (13): D-glutamine and D-glutamate metabolism; (14): pyruvate metabolism; (15): glycolysis/glycohydrogenation; (16): taurine and hypotaurine metabolism. The graph’s <span class="html-italic">X</span>-axis represents the pathway influence factor, and the <span class="html-italic">Y</span>-axis shows the enrichment analysis’s <span class="html-italic">p</span>-value. The bigger the circle, the more influential it is; the darker the circle’s color, the lower the <span class="html-italic">p</span>-value and the more significant the enrichment.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Subjects
2.2. Cardiorespiratory Fitness Testing and Grouping
2.3. Blood Sample Collection
2.4. Metabolite Extraction
2.5. Protein Precipitation
2.6. Sample Labeling
2.7. Sample Mixture
2.8. Analysis Condition and Data Quality Control and Metabolite Identification Results
2.9. Statistical Analysis
3. Results
3.1. Subjects
3.2. PLS-DA
3.3. OPLS-DA
3.4. Volcano Plot Analysis
3.5. Venn Diagram Analysis
3.6. Metabolite Pathway Analysis
4. Discussion
4.1. Differences in Plasma Metabolites between CRF Levels and Different Degrees of Mets Risk Factors
4.2. Higher CRF Levels Reduce the Risk of MetS Risk Factors
4.3. Limitation
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Name | Conditions of Association |
---|---|
chromatograph | Agilent 1290 Ultra High Performance Liquid Chromatography−6546 Quadrupole-Time of Flight Mass Spectrometer |
column | Agilent eclipse plus reversed-phase C18 column (150 mm × 2.1 mm, 1.8 µm particle size) |
mobile phase A | 0.1% (v/v) Formic acid–water |
mobile phase B | 0.1% (v/v) Formic acid–acetonitrile |
Gradient elution | t = 0 min, 25% MPB; t = 10 min, 99% MPB; t = 13 min,99% MPB t = 15 min, 99% MPB; t = 15.1 min, 25% MPB; t = 18 min, 25% MPB |
Flow rate | 400 µL/min |
column temperature | 40 °C |
Scan range | m/z 220−1000 Da |
Variables | CRF Group | MS Group | |||
---|---|---|---|---|---|
LC (n = 26) | MC (n = 26) | HC (n = 27) | LM (n = 49) | HM (n = 41) | |
Age (year) | 54.19 ± 6.01 | 53.38 ± 6.33 * | 52.85 ± 6.66 | 53.63 ± 5.98 | 52.17 ± 6.55 |
Height (cm) | 160.94 ± 6.53 | 167 ± 8.09 | 168.63 ± 6.52 * | 163.12 ± 6.71 | 168.63 ± 7.59 & |
Weight (kg) | 61.42 ± 9.01 | 67.91 ± 15.63 | 76.58 ± 24.03 * | 61.26 ± 8.40 | 77.98 ± 20.88 & |
BMI (kg/m2) | 23.63 ± 2.38 | 24.11 ± 3.89 | 26.85 ± 8.33 | 22.96 ± 2.15 | 27.30 ± 6.82 & |
WC (cm) | 80.30 ± 6.93 | 85.98 ± 13.05 | 88.56 ± 8.96 * | 80.07 ± 7.00 | 90.08 ± 11.28 & |
SBP (mmHg) | 121.81 ± 17.90 | 126.69 ± 17.85 | 130.67 ± 17.70 | 122.45 ± 17.83 | 129.51 ± 15.87 |
DBP (mmHg) | 79.42 ± 12.27 | 78.42 ± 12.81 | 82.85 ± 12.61 | 77.88 ± 12.75 | 82.34 ± 10.83 |
FPG (mmol/L) | 5.39 ± 0.85 | 5.59 ± 1.43 | 5.26 ± 0.58 | 5.17 ± 0.73 | 5.67 ± 1.14 & |
TG (mmol/L) | 1.68 ± 1.07 | 1.65 ± 1.26 | 2.31 ± 2.00 | 1.30 ± 0.73 | 2.61 ± 1.78 & |
TC (mmol/L) | 5.33 ± 1.00 | 5.11 ± 1.49 | 5.01 ± 1.41 | 5.22 ± 1.25 | 4.99 ± 1.27 & |
HDL-C (mmol/L) | 1.21 ± 0.43 | 1.17 ± 0.33 | 0.92 ± 2.07 *# | 1.27 ± 0.37 | 0.88 ± 0.13 & |
LDL-C (mmol/L) | 3.57 ± 0.85 | 3.33 ± 1.27 | 3.41 ± 1.28 | 3.50 ± 1.15 | 3.29 ± 1.04 |
Hcy (μmmol/L) | 11.46 ± 4.35 | 13.41 ± 4.77 | 13.18 ± 2.33 | 11.90 ± 3.80 | 13.66 ± 4.10 & |
VO2max | 28.90 ± 1.37 | 35.78 ± 3.41 * | 43.18 ± 1.46 *# | 34.54 ± 6.21 | 38.04 ± 5.92 & |
Variables | CRF + MS Group | |||
---|---|---|---|---|
LCLM (n = 18) | LCHM (n = 8) | HCLM (n = 12) | HCHM (n = 15) | |
Age(year) | 53.39 ± 6.01 | 56.00 ± 6.00 | 54.17 ± 6.71 | 51.80 ± 6.65 |
Height(cm) | 158.78 ± 4.20 | 165.81 ± 8.38 | 168.26 ± 5.15 * | 168.93 ± 7.61 * |
Weight(kg) | 57.98 ± 6.65 | 69.16 ± 9.16 * | 67.72 ± 8.16 * | 83.66 ± 29.98 * |
BMI(kg/m2) | 22.97 ± 2.07 | 25.13 ± 2.47 * | 23.84 ± 1.82 | 29.25 ± 10.61 * |
WC(cm) | 78.54 ± 5.66 | 84.25 ± 8.24 * | 84.68 ± 7.28 * | 91.66 ± 9.18 *& |
SBP(mmHg) | 119.06 ± 18.59 | 128.00 ± 15.55 | 121.75 ± 17.92 | 137.80 ± 14.39 *& |
DBP(mmHg) | 79.17 ± 12.32 | 80.00 ± 13.00 | 77.67 ± 12.28 | 87.00 ± 11.63 |
FPG(mmol/L) | 5.44 ± 0.99 | 5.28 ± 0.43 | 4.99 ± 0.30 | 5.47 ± 0.66 & |
TG(mmol/L) | 1.44 ± 1.03 | 2.21 ± 1.02 | 1.25 ± 0.36 # | 3.15 ± 2.37 *& |
TC(mmol/L) | 5.56 ± 0.91 | 4.81 ± 1.06 | 4.95 ± 1.73 | 5.05 ± 1.16 |
HDL-C(mmol/L) | 1.33 ± 0.46 | 0.93 ± 0.08 * | 1.07 ± 0.23 | 0.82 ± 0.94 *#& |
LDL-C(mmol/L) | 3.74 ± 0.74 | 3.20 ± 1.01 | 3.50 ± 1.77 | 3.34 ± 0.77 |
Hcy(μmmol/L) | 10.74 ± 3.59 | 13.09 ± 5.67 | 13.03 ± 3.26 | 13.30 ± 1.31 * |
VO2max | 28.90 ± 1.38 | 28.90 ± 1.24 | 43.30 ± 1.71 *# | 43.08 ± 1.29 *# |
Group | R2X | R2Y | Q2 |
---|---|---|---|
LC vs. MC vs. HC | 0.144 | 0.481 | −0.063 |
LM vs.HM | 0.322 | 0.967 | 0.520 |
LCLM vs. HCLM | 0.249 | 0.981 | 0.557 |
LCHM vs. HCHM | 0.377 | 0.992 | 0.625 |
LCLM vs. LCHM | 0.365 | 0.992 | 0.715 |
HCLM vs. HCHM | 0.290 | 0.946 | 0.195 |
LCLM vs. HCHM | 0.368 | 0.997 | 0.813 |
LCHM vs. HCLM | 0.333 | 0.998 | 0.296 |
Group | R2X | R2Y | Q2 |
---|---|---|---|
LC vs. HC | 0.259 | 0.974 | 0.607 |
LC vs. MC | 0.162 | 0.930 | 0.310 |
MC vs. HC | 0.139 | 0.810 | −0.152 |
LM vs. HM | 0.415 | 0.998 | 0.520 |
LCLM vs. HCLM | 0.296 | 0.994 | 0.548 |
LCHM vs. HCHM | 0.262 | 0.979 | 0.520 |
LCLM vs. LCHM | 0.287 | 0.966 | 0.613 |
HCLM vs. HCHM | 0.321 | 0.989 | 0.028 |
LCLM vs. HCHM | 0.342 | 0.997 | 0.651 |
LCHM vs. HCLM | 0.192 | 0.958 | 0.459 |
Compounds | CRF Group | MS Group | CRF + MS Group | |||
---|---|---|---|---|---|---|
HC vs. LC | HM vs. LM | HCLM vs. LCLM | HCHM vs. LCHM | LCHM vs. LCLM | HCHM vs. LCLM | |
L-Methionine | 0.772 * | 1.446 ** | - | 0.457 * | 2.214 * | - |
γ-Aminobutyric acid | 1.284 * | 0.701 * | - | - | - | - |
2-Oxoglutaric acid | 2.204 ** | 0.824 * | 2.872 ** | 1.481 * | - | 1.784 * |
L-Arginine | 1.343 * | 0.787 * | - | 1.251 | 0.746 ** | 1.429 ** |
L-Serine | 1.268 * | 0.725 ** | - | - | 0.703 ** | 1.404 ** |
cis-Aconitic acid | 1.681 ** | 0.809 ** | 2.026 ** | - | - | 1.289 * |
L-Glutamine | 0.581 * | 1.227 * | - | 0.783 * | - | 0.824 * |
L-Valine | 0.804 * | 1.419 ** | - | 0.787 * | - | 0.759 * |
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Fei, X.; Huang, Q.; Lin, J. Plasma Metabolomics Study on the Impact of Different CRF Levels on MetS Risk Factors. Metabolites 2024, 14, 415. https://doi.org/10.3390/metabo14080415
Fei X, Huang Q, Lin J. Plasma Metabolomics Study on the Impact of Different CRF Levels on MetS Risk Factors. Metabolites. 2024; 14(8):415. https://doi.org/10.3390/metabo14080415
Chicago/Turabian StyleFei, Xiaoxiao, Qiqi Huang, and Jiashi Lin. 2024. "Plasma Metabolomics Study on the Impact of Different CRF Levels on MetS Risk Factors" Metabolites 14, no. 8: 415. https://doi.org/10.3390/metabo14080415