The Gene and Protein Expression of the Main Components of the Lipolytic System in Human Myocardium and Heart Perivascular Adipose Tissue. Effect of Coronary Atherosclerosis
Abstract
:1. Introduction
2. Results
2.1. Patient Clinical Characteristics
2.2. Myocardial and Perivascular Adipose Tissue Expression of ATGL, CGI-58, G0S2, and HSL at the Transcript (mRNA) and Protein Levels in CAD and NCAD Patients
2.2.1. Myocardium
2.2.2. Perivascular Adipose Tissue (PVAT)
2.3. Compounds Involved in Fatty Acid Metabolism i.e., β-HAD, CS, COX4/1, FAS, SREBP-1c, GPAT1, FAT/CD36, LPL, and FABP4 at the Transcript (mRNA) and Protein Levels in Myocardium and Perivascular Adipose Tissue of CAD and NCAD Patients
2.3.1. Myocardium
2.3.2. Perivascular Adipose Tissue (PVAT)
2.4. Myocardial Expression of Proteins Contributing to Inflammation, i.e., COX-2, NFκβ and 15-LO at the Protein Level of CAD and NCAD Patients
2.5. Myocardial Lipid Content and Fatty Acid Composition in CAD and NCAD Patients
2.6. Correlation of ATGL Expression and TG Content in Human Myocardium
3. Discussion
4. Materials and Methods
4.1. Subjects
4.2. Quantitative Real-Time PCR
4.3. Protein Extraction and Western Blot
4.4. The Myocardial Lipids
4.5. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ATGL | Adipose triglyceride lipase |
β-HAD | Beta-hydroxyacyl CoA dehydrogenase |
CGI-58 | Comparative gene identification 58 |
CAD | Coronary artery disease |
COX-2 | cyclooxygenase-2 |
DG | Diacylglycerols |
FABP-4 | Fatty acid-binding protein 4 |
FASN | Fatty acid synthase |
FAT/CD36 | Fatty acid translocase (cluster of differentiation 36) |
FFA | Free fatty acids |
G0S2 | G0/G1 switch protein 2 |
GPAT1 | Glycerol-3-phosphate acyltransferase 1 |
HSL | Hormone sensitive lipase |
15-LO | Lipoxygenase |
COX4/1 | Mitochondrial electron transport chain (cytochrome c oxidase subunit IV isoform 1) |
MG | Monoacylglycerol |
NFκβ | Nuclear factor kappa B |
NCAD | Patients with no atherosclerosis qualified for a valve replacement |
PVAT | Perivascular adipose tissue |
SREBP-1c | Sterol regulatory element-binding protein 1 |
TG | Triacylglycerols |
CS | Tricarboxylic acid cycle (citrate synthase) |
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Item | Control (NCAD) (n = 11) | Coronary Atherosclerosis (CAD) (n = 42) | p Value |
---|---|---|---|
Age (years) | 61.1 ± 8.3 | 63.6 ± 8.3 | 0.37 |
Sex (M/F) | 5/6 (45.4/55.6%) | 40/2 (95.3/4.7%) * | 0.008 |
BMI (kg/m2) | 26.7 ± 5.2 | 27.3 ± 3.2 | 0.68 |
Hypertension # | 5 (45.4%) | 34 (80.9%) * | 0.02 |
Hyperlipidemia | 8 (72.7%) | 32 (76.1%) | 1.0 |
History of infarction | 0 | 17 (40.4%) * | 0.012 |
Left ventricular ejection fraction (%) | 52.2 ± 6.8 | 53.1 ± 9.5 | 0.76 |
Fasting blood glucose (mg/dL) | 95.5 ± 10.8 | 96.2 ± 14.1 | 0.94 |
Creatinine clearance (ml/min) | 86.2 ± 23.6 | 91.9 ± 0.2 | 0.37 |
Total cholesterol (mg/dL) | 198.1 ± 30.2 | 183.4 ± 41.7 | 0.32 |
HDL cholesterol (mg/dL) | 53.9 ± 10.5 | 46.6 ± 11.3 | 0.065 |
LDL cholesterol (mg/dL) | 120.4 ± 17.9 | 109.9 ± 36.6 | 0.45 |
Triglyceride (mg/dL) | 112.4 ± 22.5 | 152.9 ± 83.7 | 0.14 |
C-reactive protein (mg/L) | 2.0 ± 2.0 | 4.9 ± 9.5 | 0.25 |
Hemoglobin (g/dL) | 13.3 ± 1.1 | 13.7 ± 1.4 | 0.42 |
Platelets (×103/mm3) | 195.8 ± 29.9 | 232.8 ± 49.6 * | 0.03 |
HbA1C (%) | 5.5 ± 1.1 | 5.9 ± 0.3 | 0.43 |
Alanine aminotransferase (U/L) | 25.1 ± 12.7 | 52.9 ± 42.6 * | 0.02 |
Aspartate aminotransferase (U/L) | 30.6 ± 15.7 | 39.9 ± 28.4 | 0.27 |
Medication: | |||
ACEI/ ARB | 4 (36.4%) | 40 (95.2%) * | 0.00001 |
Beta-blockers | 10 (90.9%) | 41 (97.6%) | 0.78 |
Statins | 6 (54.5%) | 41 (97.6%) * | 0.000001 |
Proton pump inhibitors | 10 (90.9%) | 39 (92.8%) | 0.83 |
Spironolaktone | 3 (27.3%) | 7 (16.6%) | 0.42 |
Furosemide | 4 (36.4%) | 8 (19.0%) | 0.22 |
Aspirin | None | 42 (100%) | 0.00 |
Composition | NCAD | CAD |
---|---|---|
Myristic acid (14:0) | 4282.14 ± 2613.1 (3.66%) | 9158.87 ± 5719.6 (3.33%) * |
Palmitic acid (16:0) | 38,589.25 ± 25,242.1 (31.9%) | 84,158.08 ± 45,434.1 (31.31%) * |
Palmitooleic acid (16:1) | 10,635.3 ± 7981.3 (8.48%) | 27,838.65 ± 16,791.5 (10.22%) * |
Stearic acid (18:0) | 5371.43 ± 2945.7 (4.95%) | 9332.44 ± 5345.3 (3.64%) * |
Oleic acid (18:1n9c) | 48,556.93 ± 35,024.1 (39.48%) | 109,021.54 ± 57,447.9 (40.86%) * |
Linoleic acid (18:2n6c) | 10,304.8 ± 8165.3 (8.69%) | 20,929.94 ± 11,997.4 (7.93%) * |
Arachidic acid (20:0) | 70.65 ± 35.7 (0.08%) | 122.86 ± 72.2 (0.05%) * |
Linolenic acid (C18:9n3) | 1855.38 ± 1434.9 (1.6%) | 3744.42 ± 2893.9 (1.4%) |
Behenic acid (22:0) | 34.93 ± 18.2 (0.04%) | 110.63 ± 57.7 (0.04%) * |
Arachidonic acid (20:4n6) | 634.53 ± 429.7 (0.59%) | 1469.99 ± 767 (0.58%) * |
Eicosapentaenoic acid (20:5n3) | 17.01 ± 11.9 (0.02%) | 116.3 ± 100.7 (0.04%) * |
Nervonic acid (24:1) | 16.52 ± 9.8 (0.02%) | 25.71 ± 14.2 (0.01%) |
Docosahexaenoic acid (22:6n3) | 498.41 ± 353.5 (0.42%) | 1292.91 ± 805.5 (0.5%) * |
Lignoceric acid (24:0) | 104.82 ± 71.7 (0.09%) | 272.75 ± 145.9 (0.11%) * |
Saturated | 48,453.23 ± 30,585.3 (40.71%) | 103,155.63 ± 56,045.7 (38.47%) * |
Unsaturated | 72,518.89 ± 51,264.1 (59.29%) | 164,439.46 ± 87,040.3 (61.53%) * |
Total | 120,972.12 ± 81,391.2 (100%) | 267,595.09 ± 140,943.4 (100%) * |
Composition | NCAD | CAD |
---|---|---|
Myristic acid (14:0) | 47.98 ± 8.3 (9.9%) | 58.16 ± 25.1 (9.4%) |
Palmitic acid (16:0) | 184.72 ± 59.6 (36.3%) | 232.98 ± 97.1 (37.4%) |
Palmitooleic acid (16:1) | 24.18 ± 12.3 (4.6%) | 40.08 ± 27.1 (6%) |
Stearic acid (18:0) | 96.22 ± 18.5 (19.8%) | 85.84 ± 19 (15%) |
Oleic acid (18:1n9c) | 81.82 ± 41.3 (15.5%) | 118.47 ± 61.2 (18.5%) |
Linoleic acid (18:2n6c) | 29.97 ± 16 (5.7%) | 39.83 ± 17 (6.4%) * |
Arachidic acid (20:0) | 1.74 ± 0.4 (0.4%) | 2.02 ± 1 (0.3%) |
Linolenic acid (C18:9n3) | 4.67 ± 2.1 (0.9%) | 4.71 ± 3 (0.7%) |
Behenic acid (22:0) | 1.28 ± 0.5 (0.3%) | 1.38 ± 0.4 (0.2%) |
Arachidonic acid (20:4n6) | 26.26 ± 13.2 (5.1%) | 22.09 ± 6.5 (4.1%) |
Eicosapentaenoic acid (20:5n3) | 1.94 ± 0.8 (0.4%) | 1.86 ± 1 (0.3%) |
Nervonic acid (24:1) | 0.82 ± 0.3 (0.2%) | 0.89 ± 0.3 (0.2%) |
Docosahexaenoic acid (22:6n3) | 3.69 ± 2.1 (0.7%) | 4.47 ± 1.7 (0.8%) |
Lignoceric acid (24:0) | 1.03 ± 0.7 (0.2%) | 2.66 ± 2.3 (0.5%) * |
Saturated | 332.98 ± 77.1 (66.9%) | 383.04 ± 134.5 (62.9%) |
Unsaturated | 173.34 ± 82.9 (33.1%) | 232.41 ± 105.2 (37.1%) |
Total | 506.32 ± 157.5 (100%) | 615.45 ± 234.7 (100%) |
Composition | NCAD | CAD |
---|---|---|
Myristic acid (14:0) | 26.68 ± 5.7 (5.9%) | 26.38 ± 12 (5.3%) |
Palmitic acid (16:0) | 167.38 ± 42.2 (36.3%) | 173.44 ± 30.1 (35.9%) |
Palmitooleic acid (16:1) | 13.63 ± 5.9 (2.9%) | 15.16 ± 5.9 (3.1%) |
Stearic acid (18:0) | 103.15 ± 19.5 (22.9%) | 102.87 ± 18.5 (21.6%) |
Oleic acid (18:1n9c) | 74.79 ± 35.1 (15.4%) | 88.02 ± 17.5 (18.3%) |
Linoleic acid (18:2n6c) | 32.77 ± 15.5 (6.7%) | 31.88 ± 11.9 (6.5%) |
Arachidic acid (20:0) | 1.46 ± 0.3 (0.3%) | 1.74 ± 0.6 (0.4%) |
Linolenic acid (C18:9n3) | 5.15 ± 2.5 (1.1%) | 3.91 ± 2.4 (0.8%) |
Behenic acid (22:0) | 0.99 ± 0.3 (0.2%) | 1.14 ± 0.5 (0.2%) |
Arachidonic acid (20:4n6) | 31.65 ± 19.2 (6.8%) | 29.96 ± 19.6 (6.1%) |
Eicosapentaenoic acid (20:5n3) | 0.93 ± 0.2 (0.2%) | 1.63 ± 1 (0.3%) * |
Nervonic acid (24:1) | 0.59 ± 0.2 (0.1%) | 0.68 ± 0.4 (0.1%) |
Docosahexaenoic acid (22:6n3) | 3.93 ± 3.3 (0.8%) | 2.55 ± 1.2 (0.5%) |
Lignoceric acid (24:0) | 1.5 ± 0.6 (0.3%) | 3.16 ± 3 (0.7%) |
Saturated | 301.17 ± 60.8 (66%) | 308.72 ± 47 (64.1%) |
Unsaturated | 163.43 ± 70.6 (34%) | 173.8 ± 36.9 (35.9%) |
Total | 463.31 ± 128.8 (100%) | 482.52 ± 75.9 (100%) |
Gene | Primer Sequence | |
---|---|---|
Forward | Reverse | |
GAPDH | 5′-AAGCCTGCCGGTGACTAAC-3′ | 5′-GTTAAAAGCAGCCCTGGTGAC-3′ |
ATGL | 5′-GCTTCCTCGGCGTCTACTAC-3′ | 5′-CAATGAACTTGGCACCAGCC-3′ |
G0S2 | 5′-ACCACAAGCATCCACCAA-3′ | 5′-GCATTTATCCTTCCTCCCTA-3′ |
CGI-58 | 5′-AGACCCAGGTTTGACAGTGATG-3′ | 5′-AGTAAGCAGCAGCCAAGAATCC-3′ |
HSL | 5′-CACGATGGGTGGAATGGTGG-3′ | 5′-ACCAGCGACTGTGTCATTGT-3′ |
FABP4 | 5′-GGGCCAGGAATTTGACGAAG-3′ | 5′-AACTCTCGTGGAAGTGACGC-3′ |
FAT/CD36 | 5′-AAGTCACTGCGACATGATTAATGG-3′ | 5′-GAACTGCAATACCTGGCTTTTCTC-3′ |
GPAT1 | 5′-AACCCCAGTATCCCGTCTTT-3′ | 5′-CAGTCACATTGGTGGCAAAC-3′ |
β-HAD | 5′-CTTGCTCCGAGAGGGAGTC-3′ | 5′-AGCTCGTAGCTGGGAGGAAC-3′ |
CS | 5′-GATTGTGCCCAATGTCCTCT-3′ | 5′-TTCATCTCCGTCATGCCATA-3′ |
COX4/1 | 5′-GGTCACGCCGATCCATATAAG-3′ | 5′-TCTGTGTGTGTACGAGCTCATGA-3′ |
LPL | 5′–GAGATTTCTCTGTATGGCACC-3′ | 5′–CTGCAAATGAGACACTTTCTC-3′ |
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Knapp, M.; Górski, J.; Lewkowicz, J.; Lisowska, A.; Gil, M.; Wójcik, B.; Hirnle, T.; Chabowski, A.; Mikłosz, A. The Gene and Protein Expression of the Main Components of the Lipolytic System in Human Myocardium and Heart Perivascular Adipose Tissue. Effect of Coronary Atherosclerosis. Int. J. Mol. Sci. 2020, 21, 737. https://doi.org/10.3390/ijms21030737
Knapp M, Górski J, Lewkowicz J, Lisowska A, Gil M, Wójcik B, Hirnle T, Chabowski A, Mikłosz A. The Gene and Protein Expression of the Main Components of the Lipolytic System in Human Myocardium and Heart Perivascular Adipose Tissue. Effect of Coronary Atherosclerosis. International Journal of Molecular Sciences. 2020; 21(3):737. https://doi.org/10.3390/ijms21030737
Chicago/Turabian StyleKnapp, Małgorzata, Jan Górski, Janina Lewkowicz, Anna Lisowska, Monika Gil, Beata Wójcik, Tomasz Hirnle, Adrian Chabowski, and Agnieszka Mikłosz. 2020. "The Gene and Protein Expression of the Main Components of the Lipolytic System in Human Myocardium and Heart Perivascular Adipose Tissue. Effect of Coronary Atherosclerosis" International Journal of Molecular Sciences 21, no. 3: 737. https://doi.org/10.3390/ijms21030737