Plasma Metabolomics Profile of “Insulin Sensitive” Male Hypogonadism after Testosterone Replacement Therapy
<p>(<b>A</b>) Metabolic Set Enrichment Analysis (MSEA) showing the most altered metabolites revealed in the plasma of hypogonadal men before and after testosterone replacement treatment (TRT). Color intensity (white-to-red) reflects increasing statistical significance, while the circle diameter covaries with pathway impact. The graph was obtained by plotting on the <span class="html-italic">y</span>-axis the –log of <span class="html-italic">p</span>-values from pathway enrichment analysis and on the x-axis the pathway impact values derived from pathway topology analysis. (<b>B</b>) Metabolic Pathway Analysis (MetPA). All the matched pathways are displayed as circles. The color and size of each circle are based on the <span class="html-italic">p</span>-value and pathway impact value, respectively. The graph was obtained by plotting on the <span class="html-italic">y</span>-axis the −log of <span class="html-italic">p</span>-values from the pathway enrichment analysis and on the <span class="html-italic">x</span>-axis the pathway impact values derived from the pathway topology analysis.</p> "> Figure 2
<p>Metabolomic profile of glucose metabolism. (<b>A</b>) All glycolytic intermediates were upregulated after TRT. (<b>B</b>) Intermediates of pentose phosphate pathway were restored to that similar to control. (<b>C</b>) Decrease of glutathione disulphide as a marker of the improvement of oxidative stress. Deregulation of level of NAD and NADH. (<b>D</b>) The glycerol shuttle was not active to contribute to the oxidative phosphorylation pathway in the mitochondria. It was used for phospholipid synthesis. All data are shown as mean ± SEM of fold-change relative to the metabolite levels in controls. * <span class="html-italic">p</span> < 0.05.</p> "> Figure 3
<p>Metabolism involved in acetyl-CoA catabolism. Intermediates of Tricarboxylic acid (TCA) cycle measured in the plasma of hypogonadal patients. We found an overall decreased level of TCA-cycle metabolites. (<b>A</b>) Level of mevalonic acid increased after therapy, yet the concentration of cholesterol did not improve. (<b>B</b>) Acetyl-carnitine and fatty acid oxidation are significantly reduced after TRT. (<b>C</b>) Levels of energy metabolites production AMP and ATP. (<b>D</b>) Intermediates of the glutaminolysis pathway. This stepwise became downregulated after TRT. All data are shown as mean ± SEM of fold-change relative to the metabolite levels in controls. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001.</p> "> Figure 4
<p>(<b>A</b>) The downregulation of glutaminolysis led to a stop of the activation of the malate aspartate cycle, as shown by the levels of glutamate and aspartate. (<b>B</b>) Decreased levels of branched-significantly decreased. (<b>C</b>) Plasma amino acids that were significantly decreased. (<b>D</b>) Plasma amino acids that were significantly increased. (<b>E</b>) Restoration of proline and lysine involved in collagen fibers formation. All data are shown as mean ± SEM of fold-change relative to the metabolite levels in controls. * <span class="html-italic">p</span> < 0.05.</p> "> Figure 5
<p>Schematic model summarizing change in carnosine metabolism. The carnosine production from β-alanine increased in response to testosterone therapy. All data are shown as mean ± SEM of fold-change relative to the metabolite levels in controls. * <span class="html-italic">p</span> < 0.05.</p> "> Figure 6
<p>Overview of the energy production in hypogonadic insulin-sensivity male after TRT. Pyruvate produced by glycolysis is converted into lactate rather than alanine. Lactate can both be directly used as main energy source in highly oxidative cells such as brain, lung, heart or enter in liver or kidney and converted in glucose, that is used to produce energy (Cori Cycle).</p> ">
Abstract
:1. Introduction
2. Results
3. Discussion
4. Materials and Methods
4.1. Patients Samples: Study Design and Participants
4.2. Study Treatment
4.3. UHPLC-HRMS
4.4. Metabolomic Data Processing and Statistical Analysis
5. Conclusions
6. Patents
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
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Control (A) | Hypogonadic IS (B) | IS after TRT (C) | p-Value | Tukey HSD p-Value | |
---|---|---|---|---|---|
Subjects | n-20 | n-20 | n-20 | - | - |
Age | 42.54 ± 13.67 | 42.18 ± 16.01 | 42.18 ± 16.01 | - | - |
BMI (Kg/m2) | 24.91 ± 4.01 | 25.44 ± 3.04 | 25.24 ± 3.09 | 0.93 | - |
Testosterone (nmol/L) | 20.02 ± 7.47 | 6.35 ± 4.35 | 19.20 ± 9.10 | 0.0001 *** p | (A vs B) 0.001 ** p (B vs C) 0.001 ** p |
Glucose (mg/100 mL) | 84.72 ± 4.38 | 81.45 ± 12.51 | 86.90 ± 6.77 | 0.3 | - |
Insuline (mUI/L) | 7.77 ± 3.13 | 6.72 ± 2.88 | 6.99 ± 2.98 | 0.69 | - |
HOMAi | 1.79 ± 0.86 | 1.97 ± 0.67 | 1.47 ± 0.70 | 0.92 | - |
Tg (mmol/L) | 96.36 ± 51.39 | 117.90 ± 62.73 | 125.54 ± 63.18 | 0.5 | - |
Cholesterol (mmol/L) | 196.72 ± 29.18 | 212.81 ± 42.6 | 210.18 ± 53.18 | 0.6 | - |
HDL Cholesterol (mmol/L) | 53.90 ± 11.98 | 52.63 ± 15.01 | 47.45 ± 15.47 | 0.54 | - |
LDL Cholesterol (mmol/L) | 133.45 ± 33.07 | 136.36 ± 38.74 | 127.36 ± 44.31 | 0.85 | - |
Molecule | M.W. | Hypog. (%) | Post Testost. (%) |
---|---|---|---|
D-Glucose 6-Phosphate (260.14) | 260.14 | 100 | 105 |
beta-D-Fructose 1,6-bisphosphate (340.114) | 340.114 | 350 | 300 |
D-Glyceraldehyde 3-phosphate (170.06) | 170.06 | 5 | 10 |
Phosphoenolpyruvate (168.04) | 168.04 | −1 | 20 |
Lactate (90.08) | 90.08 | 10 | 280 |
D-Glucono-1,5-lactone (178.14) | 178.14 | 70 | 30 |
D-Ribulose 5-phosphate (230.11) | 230.11 | 60 | 15 |
D-Erythrose 4-phosphate (200.084) | 200.084 | −20 | −98.9 |
6-P-D-Gluconate (276.135) | 276.135 | 90 | 10 |
Sedoheptulose 1,7-bisphosphate (370.14) | 370.14 | 100 | −75 |
NADH (663.43) | 663.43 | 280 | 80 |
NAD (663.43) | 663.43 | 10 | −20 |
Glutathione disulfide (610.6) | 610.6 | 98 | −90 |
Glycerol-3-Phosphate (172.074) | 172.074 | −40 | 20 |
Dyhydroxyacetone-3P (170.06) | 170.06 | 5 | −30 |
Mevalonate (148.16) | 148.16 | 290 | 700 |
Acetyl-CoA (809.57) | 809.57 | −48 | 20 |
Acetyl-carnitine (203.236) | 203.236 | −20 | −90 |
Citrate (192.124) | 192.124 | −99.6 | −99.55 |
Oxaloacetate (132.07) | 132.07 | −18 | −1 |
Malate (134.0874) | 134.0874 | 300 | 25 |
Succinate (118.09) | 118.09 | 10 | −5 |
2-oxoglutarato (146.11) | 146.11 | 4 | 1 |
Oxalosuccinate (190.11) | 190.11 | 5 | −15 |
cis-Aconitate (174.108) | 174.108 | −20 | −100 |
Glutamate (147.13) | 147.13 | −48 | 90 |
Glutamine (146.14) | 146.14 | −5 | 2 |
AMP (347.2212) | 347.2212 | −40 | −20 |
ATP (507.18) | 507.18 | −1 | −60 |
Aspartate (133.11) | 133.11 | -60 | −55 |
Leucine/isoleucine (131.17) | 131.17 | −15 | −47 |
Valine (117.15) | 117.15 | −15 | −40 |
Tyrosine (181.19) | 181.19 | −1 | −48 |
Phenylanine (165.19) | 165.19 | −5 | −48 |
Cysteine (121.16) | 121.16 | 200 | −85 |
Tryptophan (204.23) | 204.23 | −2 | −50 |
Methionine (149.21) | 149.21 | −1 | −25 |
Alanine (89.09) | 89.09 | 280 | −55 |
Serine (105.09) | 105.09 | 299 | 60 |
Threonine (119.1192) | 119.1192 | −5 | 50 |
Asparagine (132.12) | 132.12 | −97 | −90 |
L-Arginine (174.2) | 174.2 | 60 | 200 |
Proline (115.13) | 115.13 | 50 | 47 |
L-Lysine (146.19) | 146.19 | 50 | 25 |
Histidine (155.1546) | 155.1546 | −99 | 110 |
Uracil (112.09) | 112.09 | −48 | −40 |
Carnosine (226.3) | 226.3 | −55 | −18 |
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Zolla, L.; Ceci, M. Plasma Metabolomics Profile of “Insulin Sensitive” Male Hypogonadism after Testosterone Replacement Therapy. Int. J. Mol. Sci. 2022, 23, 1916. https://doi.org/10.3390/ijms23031916
Zolla L, Ceci M. Plasma Metabolomics Profile of “Insulin Sensitive” Male Hypogonadism after Testosterone Replacement Therapy. International Journal of Molecular Sciences. 2022; 23(3):1916. https://doi.org/10.3390/ijms23031916
Chicago/Turabian StyleZolla, Lello, and Marcello Ceci. 2022. "Plasma Metabolomics Profile of “Insulin Sensitive” Male Hypogonadism after Testosterone Replacement Therapy" International Journal of Molecular Sciences 23, no. 3: 1916. https://doi.org/10.3390/ijms23031916