Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism
Abstract
:1. Introduction
2. Materials and Methods
2.1. Human Subjects Sample Collection
2.2. Isolation of PBMCs and T Cells
2.3. Immunoblotting
2.4. ROS Quantification
2.5. Mitochondrial Measures
2.6. Mitochondrial Membrane Potential
2.7. Measurement of Cellular Antioxidants
2.8. Measurement of Plasma Free Fatty Acid
2.9. Cytokine Analysis
2.10. Small Interference RNA
2.11. Statistical Analyses
3. Results
3.1. Palmitate Decreased PBMC Membrane Potential
3.2. Palmitate Altered the Cellular Redox Balance
3.3. Palmitate Lowered the Expression of Nicotinamide Nucleotide Transhydrogenase
3.4. Palmitate Increased Proinflammatory Cytokine Production by PBMCs
3.5. NNT Protein Knockdown Recapitulated Palmitate-Induced Changes in PBMCs
3.6. PBMCs from Subjects with Obesity Mimicked Characteristics of Redox Imbalance Caused by Palmitate
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Lean | Obese | |
---|---|---|
Total N | 12 | 8 |
Age, years [mean (range)] | 33 (28–45) | 39.6 (30–47) |
A1c, % [mean (range)] | 5.1 (4.5–5.6) | 5.3 (5–5.5) |
BMI, kg m−2 [mean (range)] * | 22.71 (21.1–24.4.) | 33.94 (30.4–40.8) |
Females [N (%)] | 7 (58.33%) | 4 (50%) |
Males [N (%)] | 5 (41.66%) | 4 (50%) |
Protein | Catalog # | Company |
---|---|---|
NNT | ab110352 | Abcam |
m-aconitase | ARP40390_P050 | Aviva biologicals |
Hsp60 | 12165S | Cell signaling technology |
GCLC | ARP54577_P050 | Aviva biologicals |
IDH2 | ab131263 | Abcam |
B-Actin | 3700 | Cell signaling technology |
Malic enzyme | 12399S | Cell signaling technology |
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McCambridge, G.; Agrawal, M.; Keady, A.; Kern, P.A.; Hasturk, H.; Nikolajczyk, B.S.; Bharath, L.P. Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism. Biomolecules 2019, 9, 79. https://doi.org/10.3390/biom9020079
McCambridge G, Agrawal M, Keady A, Kern PA, Hasturk H, Nikolajczyk BS, Bharath LP. Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism. Biomolecules. 2019; 9(2):79. https://doi.org/10.3390/biom9020079
Chicago/Turabian StyleMcCambridge, Grace, Madhur Agrawal, Alanna Keady, Philip A. Kern, Hatice Hasturk, Barbara S. Nikolajczyk, and Leena P. Bharath. 2019. "Saturated Fatty Acid Activates T Cell Inflammation Through a Nicotinamide Nucleotide Transhydrogenase (NNT)-Dependent Mechanism" Biomolecules 9, no. 2: 79. https://doi.org/10.3390/biom9020079