Background
Methods
Subjects
Biochemical measurements
Tissue biopsies
Total RNA extraction and quantitation
Gene expression analysis of 11β-HSD-1, GCR and CD68
Immunohistochemical staining
Determination of free fatty acids in serum and tissues
Data analysis
Results
Patient data
Obese CAD group (n = 31) | Controls (n = 16) | p value | |
---|---|---|---|
Antropometric characteristics
| |||
Age (years)
| 57.4 ± 9.1 | 58.1 ±10.4 | NS |
Female–male (%)
| 66-34 | 51-49 | NS |
BMI (kg/m
2
)
| 33.6 ± 3.1 | 30.0 ±5.2 | NS |
Waist/hip ratio
| 1.1 ± 0.0 | 0.8 ± 0.1 | NS |
Risk factors
| |||
Smoking (%)
| 44 | 66 |
0.004
|
Family history of CAD (%)
| 62 | 25 |
0.0001
|
Metabolic characteristics and indices
| |||
SBP (mm Hg)
| 135.9 ± 19.4 | 129.5 ± 30.2 | NS |
DBP (mm Hg)
| 78.5 ± 8.9 | 77.3 ± 8.1 | NS |
Total-cholesterol (mg/dl)
| 196.1 ± 31.9 | 200.1 ± 31.4 | NS |
HDL-cholesterol (mg/dl)
| 41.2 ± 9.0 | 45 ± 11.9 | NS |
LDL-cholesterol (mg/dl)
| 111.5 ± 19.0 | 142.4 ± 29.7 |
0.041
|
Triglycerides (mg/dl)
| 191.0 ± 21.5 | 201.1 ± 24.1 | NS |
Cortisol (μg/dl)
| 13.1 ± 4.5 | 8.2 ± 3.1 |
0.006
|
HOMA-IR
| 3.1 ± 1.6 | 1.9 ± 1.0 |
0.005
|
C-peptid (ng/ml)
| 2.6 ± 1.1 | 3.2 ± 1.2 | NS |
Epicardial fat (cm
3
)
| 8.8 ± 5.1 | 6.0 ± 2.5 |
0.001
|
Subcutaneous fat (cm
3
)
| 138.8 ± 44.1 | 133.1 ± 39.1 | NS |
11β-HSD-1 and GCR expressions in EAT, MAT and SAT
CD68 mRNA expressions in EAT, MAT and SAT
Correlations between gene expression levels and antropometric parameters and fat volumes
Variables | 11 β-HSD1 (EAT) | 11 β-HSD1 (MAT) | 11 β-HSD1 (SAT) | GCR (EAT) | GCR (MAT) | GCR (SAT) | ||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
r | p | r | p | r | p | r | p | r | p | r | p | |
Weight | 0.442 | 0.006 | 0.340 | 0.027 | 0.570 | 0.000 | 0.133 | 0.666 | 0.691 | 0.009 | 0.492 | 0.088 |
BMI | 0.159 | 0.363 | 0.269 | 0.118 | 0.152 | 0.384 | 0.314 | 0.07 | 0.317 | 0.060 | 0.429 | 0.009 |
Waist circumference | 0.017 | 0.926 | 0.244 | 0.171 | 0.1 | 0.58 | 0.520 | 0.002 | 0.400 | 0.019 | 0.239 | 0.189 |
Fat (kg) | 0.249 | 0.15 | 0.248 | 0.151 | 0.233 | 0.177 | 0.261 | 0.135 | 0.236 | 0.18 | 0.285 | 0.102 |
Epicardial fat (cm3) | 0.447 | 0.009 | 0.240 | 0.179 | 0.330 | 0.061 | 0.121 | 0.501 | 0.146 | 0.418 | 0.202 | 0.256 |
Subcutaneous fat (cm3) | 0.068 | 0.702 | 0.207 | 0.239 | 0.117 | 0.510 | 0.207 | 0.240 | 0.243 | 0.167 | 0.176 | 0.318 |
11 β-HSD1 (EAT) | 0.398 | 0.015 | 0.574 | 0.000 | 0.394 | 0.018 | 0.292 | 0.084 | 0.381 | 0.022 | ||
11 β-HSD1 (MAT) | 0.393 | 0.016 | 0.471 | 0.004 | 0.529 | 0.001 | 0.400 | 0.16 | ||||
11 β-HSD1 (SAT) | 0.358 | 0.032 | 0.374 | 0.025 | 0.335 | 0.046 | ||||||
GCR (EAT) | 0.496 | 0.002 | 0.389 | 0.01 | ||||||||
GCR (MAT) | 0.580 | 0.000 |
Fatty acid analysis of serum and adipose tissues
In SERUM | In EAT | In MAT | In SAT | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Fatty acids | Obese CAD group (n = 15) | Controls (n = 5) | P value | Obese CAD group (n = 15) | Controls (n = 5) | P value | Obese CAD group (n = 15) | Controls (n = 5) | P value | Obese CAD group (n = 15) | Controls (n = 5) | P value |
14:0 (myristic acid) | 32.7 ± 4.8 | 32.3 ± 7.9 | NS | 9.6 ± 0.8 | 11.9 ± 1.1 | NS | 10 ± 0.9 | 9.7 ± 1.4 | NS | 10.1 ± 0.7 | 10.4 ± 0.7 | NS |
16:0 (palmitic acid) | 625.2 ± 99.7 | 419.6 ± 121.3 | NS | 108.4 ± 7.1 | 113.9 ± 10 | NS | 103.7 ± 5.7 | 98.3 ± 11.8 | NS | 105.9 ± 7.1 | 115.6 ± 6.8 | NS |
16:1n-7 (palmitoleic acid) | 67.1 ± 12.6 | 51.6 ± 12.7 | NS | 8.3 ± 0.8 | 14.6 ± 3.3 |
0.017
| 8.9 ± 0.8 | 6.7 ± 2 | NS | 10.7 ± 1.2 | 9.8 ± 2.9 | NS |
18:0 (stearic acid) | 182.8 ± 18.3 | 169.2 ± 22.8 | NS | 26.5 ± 1.9 | 24.6 ± 4.5 | NS | 23.4 ± 1.6 | 23.9 ± 4.8 | NS | 22.5 ± 1.9 | 24 ± 3.6 | NS |
18:1n-9 (oleic acid) | 671.8 ± 115.3 | 478.4 ± 71.4 | NS | 168.4 ± 17.5 | 172.7 ± 13.5 | NS | 178 ± 14.5 | 175.9 ± 18.2 | NS | 201.2 ± 15 | 217.6 ± 10.4 | NS |
18:2n-6 (linoleic acid) | 834.7 ± 100.9 | 727.4 ± 104.3 | NS | 103.2 ± 6.8 | 105.8 ± 17.6 | NS | 106.4 ± 5.5 | 99.1 ± 17.5 | NS | 118 ± 8.8 | 117 ± 11.7 | NS |
18:3n-3 (linolenic acid) | 30.2 ± 6.4 | 23.3 ± 6.6 | NS | 1.1 ± 0.3 | 0.4 ± 0.4 | NS | 0.6 ± 0.2 | 0.1 ± 0.1 | NS | 0.6 ± 0.2 | 0.2 ± 0.2 | NS |
18:4n-3 (octatetradecanonic acid) | N.D | N.D | NS | 2.8 ± 0.3 | 1.4 ± 0.6 |
0.026
| 2.2 ± 0.4 | 0.3 ± 0.3 |
0.026
| 2.1 ± 0.4 | 2.1 ± 0.8 | NS |
20:1n-9 (eicosenoic acid) | N.D | N.D | NS | 5.9 ± 0.6 | 5 ± 0.2 | NS | 6.6 ± 0.6 | 5.1 ± 0.8 | NS | 6 ± 0.5 | 6 ± 0.6 | NS |
20:3n-6 (eicosatrienoic acid) | 40.3 ± 5.6 | 42.5 ± 8.3 | NS | 2.0 ± 0.1 | 1.3 ± 0.4 |
0.038
| 1.6 ± 0.2 | 1.4 ± 0.3 | NS | 2.2 ± 0.2 | 2.3 ± 0.6 | NS |
20:4n-6 (arachidonic acid) | 136.8 ± 18 | 131 ± 19.4 | NS | 2.5 ± 0.3 | 1.9 ± 0.5 | NS | 1.7 ± 0.2 | 1.6 ± 0.3 | NS | 2.5 ± 0.3 | 1.8 ± 0.5 | NS |
20:5n-3 (eicosapentanoic acid) | N.D | N.D | NS | N.D | N.D | NS | N.D | N.D | NS | 0.4 ± 0.3 | 0.2 ± 0,1 | NS |
22:5n-3 (dokosapentaenoic acid) | N.D | N.D | NS | 1.4 ± 0.3 | 0.9 ± 0.4 | NS | 1.2 ± 0.3 | 0.4 ± 0.2 | NS | 0.6 ± 0.2 | 0.4 ± 0.2 | NS |
22:6n-3 (docosahexaenoic acid) | N.D | N.D | NS | 2 ± 0.4 | 1.3 ± 0.7 | NS | 2.8 ± 1.7 | 0.7 ± 0.4 | NS | 4.4 ± 1.8 | 1.8 ± 0.6 | NS |
24:1n-9 (nervonic acid) | 25 ± 4.6 | 33.6 ± 11.4 | NS | 1.9 ± 0.4 | 0.6 ± 0.4 | NS | 1.6 ± 0.4 | 0.7 ± 0.3 | NS | 1.6 ± 0.3 | 1.2 ± 0.4 | NS |
n-3 (ω − 3) | 35 ± 9.5 | 40.3 ± 9.2 | NS | 7.5 ± 0.6 | 4 ± 1.3 |
0.016
| 7 ± 1.7 | 1.6 ± 0.8 | NS | 8.1 ± 1.9 | 4.8 ± 0.6 | NS |
n-6 (ω − 6) | 1011.7 ± 115.3 | 900.9 ± 125.4 | NS | 107.6 ± 7 | 109 ± 18.2 | NS | 109.6 ± 5.7 | 102 ± 17.7 | NS | 122.8 ± 9 | 121.2 ± 12 | NS |
n-9 (ω − 9) | 681.8 ± 116.2 | 491.8 ± 76.4 | NS | 176.2 ± 18 | 178.4 ± 13.9 | NS | 186.2 ± 15 | 181.6 ± 19 | NS | 209 ± 15.2 | 224.9 ± 10.7 | NS |
Saturated | 840.7 ± 119.9 | 726.4 ± 107 | NS | 144.6 ± 9.2 | 150.3 ± 15.3 | NS | 137.2 ± 16.9 | 132 ± 17.7 | NS | 138.5 ± 9.1 | 150.1 ± 10.6 | NS |
Unsaturated | 1790.9 ± 237.4 | 1476.6 ± 205 | NS | 299.8 ± 20 | 306 ± 15.3 | NS | 311.7 ± 16.9 | 292 ± 36.8 | NS | 350.4 ± 19 | 360.5 ± 21 | NS |
Ratios | ||||||||||||
16:0 / 16:1 | 11 ± 1.1 | 11.1 ± 1.1 | NS | 14.8 ± 2 | 10.1 ± 2.7 | NS | 12.6 ± 0.9 | 11.8 ± 1.8 | NS | 11 ± 1 | 12.6 ± 1.5 | NS |
18:0 / 18:1 | 0.3 ± 0.03 | 0.4 ± 0.05 | NS | 0.2 ± 0.02 | 0.14 ± 0.03 | NS | 0.14 ± 0.02 | 0.12 ± 0.02 | NS | 0.11 ± 0.01 | 0.11 ± 0.01 | NS |
18:2 / 20:4 | 6.7 ± 0.7 | 5.7 ± 0.6 | NS | 41.3 ± 3.6 | 47.1 ± 5.6 | NS | 64.4 ± 6.6 | 64.5 ± 13.5 | NS | 49.2 ± 5.3 | 49.4 ± 4.7 | NS |
n-6 / n-3 | 50.3 ± 9.6 | 24.1 ± 4.4 | NS | 16.0 ± 1.5 | 26.8 ± 6.3 |
0.023
| 19.5 ± 2.3 | 43.5 ± 15.4 |
0.010
| 19.1 ± 2.3 | 25.6 ± 4.2 | NS |
Saturated / Unsaturated | 0.5 ± 0.02 | 0.5 ± 0.02 | NS | 0.5 ± 0.03 | 0.5 ± 0.04 | NS | 0.4 ± 0.02 | 0.4 ± 0.04 | NS | 0.4 ± 0.02 | 0.42 ± 0.03 | NS |
Linear multiple regression analysis
Mediastinal adipokines mRNA expressions | Variables | R2 | Standardized β | P Value |
---|---|---|---|---|
11β–HSD-1 mRNA expression
| MAT GCR mRNA levels | 0.402 | 0.448 |
0.01
|
Cortisol | 0.233 |
0.029
| ||
C:18:4n-3 | 0.189 |
0.064
| ||
HOMA-IR | −0.190 | 0.509 | ||
GCR mRNA expression
| Weight | 0.351 | 0.131 |
0.039
|
MAT 11β–HSD-1 mRNA levels | 0.785 |
0.005
| ||
Cortisol | −0.501 |
0.018
| ||
Weight | 0.756 |
0.038
| ||
CD68 mRNA expression
| MAT 11β–HSD-1mRNA levels | 0.564 | 0.480 |
0.004
|
MAT GCR mRNA levels | 0.351 |
0.019
| ||
Cortisol | 0.250 |
0.031
| ||
Weight | 0.253 | 0.065 |