Increased Expression Profile and Functionality of TLR6 in Peripheral Blood Mononuclear Cells and Hepatocytes of Morbidly Obese Patients with Non-Alcoholic Fatty Liver Disease
Abstract
:1. Introduction
2. Results
2.1. Baseline Patient Characteristics and Histologic Analysis
2.2. Toll-Like Receptor 2 (TLR2) and TLR6 Expression Profile and Functionality in Peripheral Blood Mononuclear Cells (PBMCs)
2.3. TLR6 and TLR2 Liver Immunohistochemistry and Cytokine Expression Analysis
3. Discussion
4. Materials and Methods
4.1. Patients
4.2. TLR Expression Profile in PBMCs
4.3. Assessment of TLR Function in Circulating Monocytes
4.4. Histology Assessment
4.5. Immunohistochemistry
4.6. Cytokine Gene Expression Analysis
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
NAFLD | Non-alcoholic fatty liver disease |
NASH | Non-alcoholic steatohepatitis |
TLR | Toll-like receptor |
PBMCs | Peripheral blood mononuclear cells |
MO | Morbidly obese |
AST | aspartate aminotransferase |
ALT | alanine aminotransferase |
GGT | γ-glutamyl transpeptidase |
AP | alkaline phosphatase |
HOMA | homeostasis model assessment |
MFI | Mean fluorescence intensity |
IL | Interleukin |
TNF | Tumor necrosis factor |
NAFL | Non-alcoholic fatty liver |
IHC | Immunohistochemistry |
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Patients’ Characteristics | Overall (n = 40) | Non-NASH (n = 17) | NASH (n = 17) | p |
---|---|---|---|---|
Male/female | 15/25 (37.5/62.5) | 3/14 (17.6/82.4) | 11/6 (64.7/35.3) | 0.007 |
Age | 43.78 (10.33) | 41.00 (9.01) | 42.76 (10.9) | NS |
Metabolic syndrome | 23 (57.5) | 6 (35.3) | 13 (76.5) | 0.02 |
Obstructive sleep apnea | 27 (67.5) | 8 (47.1) | 15 (88.2) | 0.01 |
BMI (kg/m2) | 48.17 (4.93) | 49.52 (5.7) | 47.79 (3.2) | NS |
Waist circumference (cm) | 139.48 (11.24) | 135.94 (9.5) | 143.47 (12.4) | NS |
Hip circumference (cm) | 144.61 (10.41) | 146.47 (11.9) | 143.62 (8.1) | NS |
Systolic blood pressure (mm Hg) | 136.27 (13.99) | 132.81 (9.2) | 142.56 (16.4) | 0.05 |
Diastolic blood pressure (mm Hg) | 82.89 (14.44) | 83.25 (15.2) | 82.94 (16.2) | NS |
LDL-Cholesterol (mg/dL) | 102.38 (33.11) | 109.13 (31.9) | 91.44 (31.7) | NS |
HDL-Cholesterol (mg/dL) | 40.35 (10.14) | 43.80 (10.5) | 37.31 (8.8) | NS |
Total cholesterol (mg/dL) | 179.97 (40.38) | 184.24 (39.4) | 172.63 (41.2) | NS |
Tryglicerides (mg/dL) | 167.57 (93.19) | 128.44 (62.9) | 208.30 (107.5) | 0.02 |
Homocysteine (µmol/L) | 11.29 (3.49) | 11.72 (4.6) | 10.84 (2.64) | NS |
cCRP (mg/L) | 9.82 (10.73) | 13.34 (15.1) | 8.08 (5.4) | NS |
AST (U/L) | 28.11 (12.40) | 23.00 (7.6) | 34.88 (14.5) | 0.007 |
ALT (U/L) | 33.51 (18.52) | 23.53 (7.5) | 45.59 (21.8) | 0.001 |
GGT (U/L) | 32.08 (21.12) | 29.50 (16.4) | 35.41 (26.3) | NS |
Alkaline phosphatase (U/L) | 70.89 (17.86) | 72.69 (18.6) | 68.65 (18.8) | NS |
Bilirubin (mg/dL) | 0.77 (0.31) | 0.67 (0.2) | 0.85 (0.4) | NS |
Albumin (mg/dL) | 4.16 (0.27) | 4.18 (0.2) | 4.15 (0.3) | NS |
Ferritin (mg/dL) | 159.60 (160.38) | 99.88 (79.3) | 261.36 (201.5) | 0.02 |
HbA1c (%) | 6.22 (1.58) | 5.48 (0.3) | 6.99 (2.1) | 0.008 |
Platelets (103/µL) | 240.87 (62.04) | 269.35 (58.9) | 213.53 (51.4) | 0.006 |
Prothrombin time (%) | 77.79 (9.08) | 75.71 (7.8) | 78.82 (7.1) | NS |
HOMA index | 4.01 (4.24) | 3.29 (1.5) | 5.76 (5.7) | NS |
25-OH-Vit D (ng/mL) | 15.95 (5.74) | 15.18 (6.0) | 15.47 (4.0) | NS |
Liver Histology according to Brunt’s Classification | |
---|---|
Steatosis grade (%) | |
0: minimal or <5% | 10 (25) |
1: 5%–33% | 14 (35) |
2: >33%–66% | 3 (7.5) |
3: >66% | 7 (17.5) |
Lobular inflammation grade (%) | |
0: no foci | 15 (44.1) |
1: <2 foci per 200 × field | 18 (52.9) |
2: 2–4 foci per 200 × field | 0 (0) |
3: >4 foci per 200 × field | 1 (2.9) |
Ballooning grade (%) | |
0: none | 23 (67.6) |
1: few balloon cells | 4 (11.8) |
2: many cells, prominent ballooning | 7 (20.6) |
NAFLD activity index score (NAS) (%) | |
0: normal liver histology | 5 (14.7) |
1–2: not steatohepatitis | 12 (35.3) |
3–4: possible or borderline steatohepatitis | 14 (41.2) |
5–8: definite steatohepatitis | 3 (8.8) |
Fibrosis grade (%) | |
F0: none | 5 (14.7) |
F1: perisinusoidal or periportal | 21 (61.8) |
F2: perisinusoidal and portal/periportal | 7 (20.6) |
F3: bridging fibrosis | 1 (2.9) |
F4: cirrhosis | 0 (0) |
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Arias-Loste, M.T.; Iruzubieta, P.; Puente, Á.; Ramos, D.; Santa Cruz, C.; Estébanez, Á.; Llerena, S.; Alonso-Martín, C.; San Segundo, D.; Álvarez, L.; et al. Increased Expression Profile and Functionality of TLR6 in Peripheral Blood Mononuclear Cells and Hepatocytes of Morbidly Obese Patients with Non-Alcoholic Fatty Liver Disease. Int. J. Mol. Sci. 2016, 17, 1878. https://doi.org/10.3390/ijms17111878
Arias-Loste MT, Iruzubieta P, Puente Á, Ramos D, Santa Cruz C, Estébanez Á, Llerena S, Alonso-Martín C, San Segundo D, Álvarez L, et al. Increased Expression Profile and Functionality of TLR6 in Peripheral Blood Mononuclear Cells and Hepatocytes of Morbidly Obese Patients with Non-Alcoholic Fatty Liver Disease. International Journal of Molecular Sciences. 2016; 17(11):1878. https://doi.org/10.3390/ijms17111878
Chicago/Turabian StyleArias-Loste, María Teresa, Paula Iruzubieta, Ángela Puente, David Ramos, Carolina Santa Cruz, Ángel Estébanez, Susana Llerena, Carmen Alonso-Martín, David San Segundo, Lorena Álvarez, and et al. 2016. "Increased Expression Profile and Functionality of TLR6 in Peripheral Blood Mononuclear Cells and Hepatocytes of Morbidly Obese Patients with Non-Alcoholic Fatty Liver Disease" International Journal of Molecular Sciences 17, no. 11: 1878. https://doi.org/10.3390/ijms17111878