TIGAR Deficiency Blunts Angiotensin-II-Induced Cardiac Hypertrophy in Mice
<p>Effect of TIGAR deficiency on systolic function in Ang-II-induced hypertension. (<b>A</b>) Schematic of the experimental design. (<b>B</b>) Systolic blood pressure after four weeks of Ang-II infusion measured by the tail-cuff method. (<b>C</b>) The representative echocardiographic images of WT and TIGAR KO mice subjected to either vehicle or Ang-II infusion for four weeks. (<b>D</b>–<b>F</b>) Left ventricular (LV) ejection fraction (EF), fractional shortening (FS), and LV mass measured by echocardiography in the indicated groups (n = 8). * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 2
<p>Effect of TIGAR deficiency on diastolic function in Ang-II-induced hypertension. (<b>A</b>) The representative pulsed-wave Doppler and tissue Doppler images from an apical four-chamber view of WT and TIGAR KO mice infused with either vehicle or Ang-II for four weeks. (<b>B</b>) The isovolumic relaxation time (IVRT) was increased in the WT and TIGAR KO mice after Ang-II infusion for four weeks. (<b>C</b>) The myocardial performance index (MPI) was also increased in the WT and TIGAR KO mice after Ang-II infusion for four weeks. (<b>D</b>) The representative pulsed-wave Doppler images of the proximal left coronary arteries of WT and TIGAR KO mice infused with either vehicle or Ang-II for four weeks. (<b>E</b>) The coronary flow reserve (CFR) was not affected by Ang-II infusion. n = 8, * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 3
<p>Knockout of TIGAR attenuates Ang-II-induced cardiac hypertrophy. (<b>A</b>) The ratio of heart weight to tibia length in the indicated groups (n = 8). (<b>B</b>,<b>C</b>) The representative images of wheat germ agglutinin (WGA)–stained frozen heart sections and cardiomyocyte hypertrophy were assessed by cross-sectional areas in the indicated groups (n = 4–5). A minimum of 100 cardiomyocytes from each LV section of each mouse were measured. Bar = 50 μm. (<b>D</b>,<b>E</b>) The representative images of Picrosirius red-stained paraffin-embedded heart sections and quantification of the percentage of interstitial fibrosis area in the indicated groups (n = 3–4). Bar = 50 μm. (<b>F</b>–<b>H</b>) The representative images of Picrosirius red-stained paraffin-embedded heart sections show coronary arteries and perivascular fibrosis and quantification of perivascular fibrosis index and vascular remodeling in the indicated groups. Bar = 25 μm. n = 3–4, * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01, *** <span class="html-italic">p</span> < 0.001, **** <span class="html-italic">p</span> < 0.0001.</p> "> Figure 4
<p>Knockout of TIGAR increases glycolytic enzyme PFK-1 and glucose transporter Glut-4. (<b>A</b>–<b>F</b>) Representative immunoblots and quantitative analysis of TIGAR, PFKFB3, PFK-1, Glut-1, Glut-4, and corresponding GAPDH or β-tubulin in the indicated groups. The expression of PFK-1 and Glut-4 decreased in the WT mice after Ang-II infusion but not in the TIGAR KO mice. n = 3–4, * <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 5
<p>Effect of TIGAR deficiency on glycolytic enzymes in Ang-II-induced hypertension. (<b>A</b>) Cardiac F2,6-BP level was determined the coupled-enzymatic assay and expressed as the fold change to the WT sham group. n = 4. (<b>B</b>) Cardiac PFK-1 activity was determined by the coupled-enzymatic assay and expressed as the OD<sub>340nm</sub>/min/mg protein. n = 7–8. * <span class="html-italic">p</span> < 0.05, ** <span class="html-italic">p</span> < 0.01.</p> ">
Abstract
:1. Introduction
2. Results
2.1. TIGAR Deficiency Does Not Affect Systolic Function in Ang-II-Induced Hypertension
2.2. TIGAR Deficiency Does Not Affect Diastolic Function in Ang-II-Induced Hypertension
2.3. Knockout of TIGAR Attenuates Ang-II-Induced Cardiac Hypertrophy
2.4. Knockout of TIGAR Increases Glycolytic Enzyme PFK-1 and Glucose Transporter Glut-4 Expression
3. Discussion
4. Materials and Methods
4.1. Mice
4.2. Measurement of Blood Pressure
4.3. Echocardiography
4.4. Tissue F2,6-BP Assay
4.5. Phosphofructokinase Activity Assay
4.6. Histology and Immunofluorescence
4.7. Immunoblot Analysis
4.8. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Parameters | WT | TIGAR KO | ||
---|---|---|---|---|
Vehicle (n = 8) | Ang-II (n = 8) | Vehicle (n = 8) | Ang-II (n = 8) | |
Heart Rate | 449 ± 10 | 479 ± 4 * | 465 ± 4 | 478 ± 6 |
Diameter; s (mm) | 2.71 ± 0.09 | 2.57 ± 0.11 | 2.60 ± 0.07 | 2.42 ± 0.17 |
Diameter; d (mm) | 4.00 ± 0.11 | 3.56 ± 0.13 | 3.88 ± 0.06 | 3.40 ± 0.15 # |
Volume; s (µL) | 27.65 ± 2.14 | 24.56 ± 2.52 | 24.99 ± 1.57 | 22.28 ± 4.60 |
Volume; d (µL) | 70.50 ± 4.23 | 53.95 ± 4.73 | 65.48 ± 2.41 | 48.59 ± 5.64 # |
SV (µL) | 42.85 ± 2.57 | 29.39 ± 2.96 * | 40.49 ± 1.24 | 26.31 ± 1.44 # |
CO (mL/min) | 19.17 ± 1.21 | 14.14 ± 1.50 * | 18.84 ± 0.60 | 12.59 ± 0.70 # |
LVAW; s (mm) | 1.12 ± 0.03 | 1.23 ± 0.04 | 1.15 ± 0.03 | 1.11 ± 0.04 |
LVAW; d (mm) | 0.76 ± 0.02 | 0.92 ± 0.03 * | 0.79 ± 0.03 | 0.81 ± 0.03 |
LVPW; s (mm) | 1.07 ± 0.04 | 1.20 ± 0.06 | 1.02 ± 0.03 | 1.07 ± 0.05 |
LVPW; d (mm) | 0.75 ± 0.05 | 0.96 ± 0.06 * | 0.66 ± 0.02 | 0.84 ± 0.04 # |
Diastolic Index | WT | TIGAR KO | ||
---|---|---|---|---|
Vehicle (n = 8) | Ang-II (n = 8) | Vehicle (n = 8) | Ang-II (n = 8) | |
E (cm/s) | 84.37 ± 2.57 | 75.89 ± 3.12 | 82.28 ± 2.11 | 68.39 ± 3.85 # |
A (cm/s) | 61.58 ± 2.30 | 55.34 ± 3.28 | 61.58 ± 2.81 | 47.55 ± 2.87 # |
E/A ratio | 1.38 ± 0.06 | 1.41 ± 31.83 | 1.35 ± 0.06 | 1.51 ± 0.16 |
e’ (cm/s) | 2.95 ± 0.13 | 2.54 ± 0.26 | 2.83 ± 0.05 | 2.20 ± 0.15 # |
a’ (cm/s) | 2.51 ± 0.09 | 2.18 ± 0.24 | 2.50 ± 0.06 | 1.75 ± 0.10 # |
E/e’ ratio | 29.22 ± 1.83 | 31.83 ± 3.10 | 29.16 ± 0.95 | 31.75 ± 2.05 |
IVCT (ms) | 8.92 ± 0.51 | 8.88 ± 0.74 | 9.72 ± 0.52 | 9.28 ± 0.39 |
AET (ms) | 49.50 ± 0.90 | 49.33 ± 1.52 | 50.38 ± 1.18 | 52.06 ± 2.89 |
Coronary Flow Velocity | WT | TIGAR KO | ||
---|---|---|---|---|
Vehicle (n = 8) | Ang-II (n = 8) | Vehicle (n = 8) | Ang-II (n = 8) | |
Sbaseline (cm/s) | 5.67 ± 0.44 | 9.13 ± 1.05 | 6.63 ± 0.54 | 11.46 ± 3.65 |
Dbaseline (cm/s) | 28.58 ± 1.65 | 36.33 ± 3.19 | 30.59 ± 1.45 | 33.80 ± 3.41 |
Shyperremia (cm/s) | 25.64 ± 1.78 | 35.93 ± 3.41 * | 28.70 ± 1.83 | 30.08 ± 3.19 |
Dhyperremia (cm/s) | 83.73 ± 2.92 | 88.67 ± 4.37 | 82.24 ± 2.76 | 75.32 ± 4.37 |
Primary Antibody | Catalog # | Host | Vender | MW (kDa) | Dilution |
---|---|---|---|---|---|
GAPDH | #2118 | rabbit | Cell Signaling, Danvers, MA, USA | 37 | 1:5000 |
Glut-1 | NB110-39113 | rabbit | Novus Biologicals, Centennial, CO, USA | 55 | 1:1000 |
Glut-4 | #2213 | mouse | Cell Signaling, Danvers, MA, USA | 50 | 1:1000 |
PFK-1 | sc-377346 | rabbit | Santa Cruz, Dallas, TX, USA | 85 | 1:1000 |
PFKFB3 | ab181861 | rabbit | Abcam, Boston, MA, USA | 58 | 1:1000 |
TIGAR | sc-67273 | rabbit | Santa Cruz, Dallas, TX, USA | 30 | 1:1000 |
β-tubulin | #86298 | mouse | Cell Signaling, Danvers, MA, USA | 55 | 1:2000 |
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He, X.; Williams, Q.A.; Cantrell, A.C.; Besanson, J.; Zeng, H.; Chen, J.-X. TIGAR Deficiency Blunts Angiotensin-II-Induced Cardiac Hypertrophy in Mice. Int. J. Mol. Sci. 2024, 25, 2433. https://doi.org/10.3390/ijms25042433
He X, Williams QA, Cantrell AC, Besanson J, Zeng H, Chen J-X. TIGAR Deficiency Blunts Angiotensin-II-Induced Cardiac Hypertrophy in Mice. International Journal of Molecular Sciences. 2024; 25(4):2433. https://doi.org/10.3390/ijms25042433
Chicago/Turabian StyleHe, Xiaochen, Quinesha A. Williams, Aubrey C. Cantrell, Jessie Besanson, Heng Zeng, and Jian-Xiong Chen. 2024. "TIGAR Deficiency Blunts Angiotensin-II-Induced Cardiac Hypertrophy in Mice" International Journal of Molecular Sciences 25, no. 4: 2433. https://doi.org/10.3390/ijms25042433