Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay
Abstract
:1. Introduction
2. Experimental Section
2.1. Development of an Assay for LP-Z Particle Quantification
2.2. Evaluation of LP-Z Assay Performance
2.3. Isolation of LP-Z Particles from High Bilirubin Samples
2.4. Agarose Gel Electrophoresis and Staining
2.5. Lipid and ApoB Content of Isolated Lipoproteins
2.6. Targeted Lipidomic and Metabolomic Analysis
2.7. Protein Delipidation and Solubilization
2.8. Tryptic Protein Digestion
2.9. Proteomic Analysis
2.10. Polyacrylamide Gel Electrophoresis
2.11. Quantification of LP-Z in Samples from Subjects with Liver Disease or Controls
2.12. Statistical Analysis
3. Results
3.1. Analytical Performance of the LP-Z NMR Assay
3.2. Characterization of Isolated LP-Z Particles
3.3. Quantification of LPZ Particles in Serum of Subjects with Various Liver Diseases
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Lipid Related Parameters | Normal Control 1 | Normal Control 2 | High Bilirubin 1 | High Bilirubin 2 | High Bilirubin 3 | High Bilirubin 4 |
---|---|---|---|---|---|---|
Bilirubin (mg/dL) | ND | ND | 22.2 | 13.4 | 18.9 | 24.6 |
TG (mg/dL) | 48.0 | 59.9 | 145.9 | 347.8 | 443.8 | 771.2 |
TC (mg/dL) | 121.3 | 189.3 | 90.7 | 194.8 | 265.6 | 343.5 |
LDL-C (mg/dL) | 77.1 | 82.9 | 80.1 | 169.3 | 98.7 | 84.3 |
HDL-C (mg/dL) | 38.4 | 102.3 | 6.4 | 4.9 | 7.7 | 6.7 |
Apo A-I (mg/dL) | 102.0 | 169.4 | 19.7 | 14.7 | 23.1 | 21.6 |
Apo B (mg/dL) | 60.8 | 57.1 | 175.0 | 364.0 | 368.3 | 304.1 |
TRL-P (nmol/L) | 26.8 | 18.6 | 16.3 | 81.5 | 985.8 | 651.8 |
VL-TRL-P (nmol/L) | 0.1 | 0.0 | 0.0 | 0.0 | 0.0 | 3.5 |
L-TRL-P (nmol/L) | 0.0 | 0.0 | 0.0 | 0.0 | 2.2 | 6.6 |
M-TRL-P (nmol/L) | 4.5 | 1.5 | 16.3 | 81.5 | 63.0 | 369.2 |
S-TRL-P (nmol/L) | 22.2 | 17.1 | 0.0 | 0.0 | 0.0 | 0.0 |
VS-TRL-P (nmol/L) | 0.0 | 0.0 | 0.0 | 0.0 | 920.6 | 272.5 |
LDL-P (nmol/L) | 1112 | 1170 | 3427 | 7026 | 5693 | 4531 |
L-LDL-P (nmol/L) | 220.9 | 0.0 | 0.0 | 0.0 | 0.0 | 123.8 |
M-LDL-P (nmol/L) | 695.3 | 429.3 | 0.0 | 0.0 | 0.0 | 0.0 |
S-LDL-P (nmol/L) | 195.6 | 740.2 | 458.8 | 1085.2 | 0.0 | 0.0 |
LP-Z (nmol/L) | 0.0 | 0.0 | 2968 | 5941 | 5693 | 4407 |
HDL-P (µmol/L) | 19.4 | 20.8 | 3.8 | 4.9 | 7.7 | 6.3 |
L-HDL-P (µmol/L) | 0.9 | 7.2 | 0.3 | 0.0 | 0.0 | 0.0 |
M-HDL-P (µmol/L) | 4.1 | 6.2 | 0.0 | 0.0 | 0.0 | 0.0 |
S-HDL-P (µmol/L) | 14.4 | 7.4 | 3.6 | 4.9 | 7.7 | 6.3 |
TRLZ (nm) | 36.4 | 37.7 | 42.0 | 40.3 | 50.1 | 51.0 |
LDLZ (nm) | 21.1 | 20.1 | 19.0 | 19.0 | 19.9 | 22.4 |
HDLZ (nm) | 8.8 | 10.6 | 8.3 | 7.4 | 7.4 | 7.5 |
Mass % Composition | Normal Control 1 | Normal Control 2 | High Bilirubin 1 | High Bilirubin 2 | High Bilirubin 3 | High Bilirubin 4 |
---|---|---|---|---|---|---|
Protein | 21.9 | 20.5 | 21.4 | 22.6 | 23.9 | 22.2 |
Phospholipids | 20.5 | 21.3 | 26.6 | 23.3 | 23.9 | 26.7 |
Triglycerides (TG) | 11.4 | 15.8 | 29.2 | 44.1 | 32.7 | 34.6 |
Free cholesterol (FC) | 7.4 | 5.4 | 8.6 | 6.2 | 8.6 | 9.2 |
Cholesteryl esters (CE) | 38.8 | 37.0 | 14.2 | 3.8 | 10.8 | 7.4 |
FC/CE ratio | 0.2 | 0.1 | 0.6 | 1.6 | 0.8 | 1.2 |
TG/CE ratio | 0.3 | 0.4 | 2.1 | 11.6 | 3.0 | 4.7 |
Subjects (# of Samples Tested) | TG (mg/dL) | TC (mg/dL) | LP-Z (nmol/L) |
---|---|---|---|
Normal healthy adults (769) | 148 ± 94 | 186 ± 36 | 0.0 |
Primary biliary cholangitis (PBC) (11) | 113 ± 30 | 173 ± 21 | 0.0 |
Primary sclerosing cholangitis (PSC) (11) | 131 ± 61 | 179 ± 48 | 0.0 |
Autoimmune hepatitis (AIH) (19) | 116 ± 64 | 170 ± 48 | 0.0 |
Alcoholic liver disease (ALD) (47) | 99 ± 72 | 133 ± 46 | 1550 ± 1257 * |
T2DM and metabolic syndrome (138) | 172 ± 124 | 192 ± 31 | 0.0 |
Hypertriglyceridemia (≥500 mg/dL TG) (1970) | 730 ± 266 | 273 ± 58 | 1690 ± 772 ** |
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Bedi, S.; Garcia, E.; Jeyarajah, E.J.; Shalaurova, I.; Perez-Matos, M.C.; Jiang, Z.G.; Dullaart, R.P.F.; Matyus, S.P.; Kirk, W.J.; Otvos, J.D.; et al. Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay. J. Clin. Med. 2020, 9, 2915. https://doi.org/10.3390/jcm9092915
Bedi S, Garcia E, Jeyarajah EJ, Shalaurova I, Perez-Matos MC, Jiang ZG, Dullaart RPF, Matyus SP, Kirk WJ, Otvos JD, et al. Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay. Journal of Clinical Medicine. 2020; 9(9):2915. https://doi.org/10.3390/jcm9092915
Chicago/Turabian StyleBedi, Shimpi, Erwin Garcia, Elias J. Jeyarajah, Irina Shalaurova, Maria Camila Perez-Matos, Z. Gordon Jiang, Robin P. F. Dullaart, Steven P. Matyus, William J. Kirk, James D. Otvos, and et al. 2020. "Characterization of LP-Z Lipoprotein Particles and Quantification in Subjects with Liver Disease Using a Newly Developed NMR-Based Assay" Journal of Clinical Medicine 9, no. 9: 2915. https://doi.org/10.3390/jcm9092915