GPAT Gene Silencing in Muscle Reduces Diacylglycerols Content and Improves Insulin Action in Diet-Induced Insulin Resistance
Abstract
:1. Introduction
2. Results
2.1. Insulin Sensitivity
2.2. Plasma Free Fatty Acids (FFA) Concentration
2.3. Fatty Acid Transporters
2.3.1. CD36
2.3.2. FABPpm
2.3.3. FATP1
2.4. Skeletal Muscle Short and Long Chain Acyl-CoA
2.5. Skeletal Muscle CPT1B Content.
2.6. Skeletal Muscle Short and Long-Chain Acyl-Carnitine Content
2.7. Skeletal Muscle mRNA and Protein Content of GPAT
2.8. Skeletal Muscle DAG Level
2.9. Skeletal Muscle TAG Level
2.10. Skeletal Muscle Ceramide Content
2.11. Insulin Cascade Activation/Inhibition
2.11.1. IR
2.11.2. IRS-1
2.11.3. PI3K
2.11.4. Akt Phosphorylation
2.11.5. AS160
2.12. Glucotransporter 4 (GLUT4)
2.13. Skeletal Muscle Glucose Uptake
3. Discussion
4. Materials and Methods
4.1. Plasmids and In Vivo Electroporation
4.2. Lipid Measurements
4.2.1. Plasma FFA
4.2.2. Sphingolipids
4.2.3. Diacylglycerols
4.2.4. Malonyl-CoA and Long-Chain Acyl-CoA
4.2.5. Acyl-carnitines
4.2.6. Triacylglycerols
4.3. Western Blotting
4.4. Real-Time PCR
4.5. Insulin-Stimulated Glucose Uptake
4.6. Plasma Insulin and Glucose Concentration
4.7. Oral Glucose Tolerance Test (OGTT)
4.8. Insulin Tolerance Test (ITT)
4.9. HOMA-IR (Homeostatic Model Assessment of Insulin Resistance)
4.10. Protein Concentration
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
HFD | High-fat diet |
GPAT | Glycerol-3-phosphate acyltransferase |
DAG | Diacylglycerols |
T2D | Type 2 diabetes |
FFA | Free fatty acids |
FA | Fatty acids |
FAT/CD36 | Fatty acid translocase |
FABPpm | Fatty acid binding protein |
FATP1–6 | Fatty acid transport protein family |
ACS | Acyl-Co synthase |
LCA-CoA | Long chain acyl-CoA |
TAG | Triacylglycerols |
Cer | Ceramides |
CPT1 | Carnitine palmitoyltransferase 1 |
IMCL | Intramyocellular lipids |
PKC | Protein kinase C |
Akt/PKB | Protein kinase B |
PPA2 | Phosphatase A2 |
LC/MS | Liquid chromatography mass spectrometry |
HOMA-IR | Homeostatic model assessment |
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Control | HFD | |
---|---|---|
Glucose (mg/dL) | 109 ± 14 | 184 ± 22 a |
Insulin (μU/mL) | 25.1 ± 2.9 | 41.3 ± 2.2 a |
OGTT AUC (×103) | 2.32 ± 0.12 | 4.27 ± 0.21 a |
ITT AUC (×103) | 0.78 ± 0.04 | 1.86 ± 0.12 a |
HOMA-IR | 1.12 ± 0.09 | 3.14 ± 0.45 a |
16:0/16:0 | 16:0/18:0 | 16:0/18:1 | 16:0/18:2 | 18:0/18:0 | 18:0/18:1 | 18:0/18:2 | 18:1/18:1 | 18:2/18:2 | 18:0/20:0 | |
---|---|---|---|---|---|---|---|---|---|---|
Control | 13.41 ± 1.87 | 83.47 ± 11.87 | 32.18 ± 4.97 | 38.09 ± 5.91 | 2.10 ± 0.23 | 21.88 ± 3.11 | 0.96 ± 0.19 | 20.98 ± 2.31 | 24.91 ± 2.63 | 3.85 ± 0.58 |
HFD(+GPAT) | 16.15 ± 2.47 a | 144.52 ± 16.79 a | 39.36 ± 4.67 a | 66.52 ± 11.60 a | 2.92 ± 0.23 a | 22.90 ± 3.65 | 1.98 ± 0.33 a | 23.37 ± 2.21 a | 34.00 ± 5.15 a | 7.23 ± 0.96 a |
HFD(-GPAT) | 11.52 ± 1.11 b | 59.86 ± 10.31 a,b | 24.15 ± 1.21 a,b | 21.31 ± 3.68 a,b | 0.91 ± 0.11 a,b | 16.74 ± 2.42 a,b | 1.53 ± 0.13 a,b | 16.73 ± 1.25 a,b | 18.68 ± 1.72 a,b | 4.90 ± 0.50 a,b |
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Kojta, I.; Zabielski, P.; Roszczyc-Owsiejczuk, K.; Imierska, M.; Sokołowska, E.; Błachnio-Zabielska, A. GPAT Gene Silencing in Muscle Reduces Diacylglycerols Content and Improves Insulin Action in Diet-Induced Insulin Resistance. Int. J. Mol. Sci. 2020, 21, 7369. https://doi.org/10.3390/ijms21197369
Kojta I, Zabielski P, Roszczyc-Owsiejczuk K, Imierska M, Sokołowska E, Błachnio-Zabielska A. GPAT Gene Silencing in Muscle Reduces Diacylglycerols Content and Improves Insulin Action in Diet-Induced Insulin Resistance. International Journal of Molecular Sciences. 2020; 21(19):7369. https://doi.org/10.3390/ijms21197369
Chicago/Turabian StyleKojta, Iwona, Piotr Zabielski, Kamila Roszczyc-Owsiejczuk, Monika Imierska, Emilia Sokołowska, and Agnieszka Błachnio-Zabielska. 2020. "GPAT Gene Silencing in Muscle Reduces Diacylglycerols Content and Improves Insulin Action in Diet-Induced Insulin Resistance" International Journal of Molecular Sciences 21, no. 19: 7369. https://doi.org/10.3390/ijms21197369