Introduction
Subject and method
Ethics approval and consent to participate
Sampling of blood
Biochemical analysis
Immunochemical assays
RNA extraction, cDNA synthesis and real time PCR for ICAM-1 mRNA gene expression
Statistical analysis
Results
Clinical and laboratory characteristics of the study subjects
Control group (mean ± SD) (n = 120) | PCO patients (mean ± SD) (n = 180) | P | |
---|---|---|---|
Age (years) | 31.38 ± 7.88 | 31.95 ± 7.42 | 0.527 |
Systolic blood pressure (mm Hg) | 127.6 ± 5.62 | 131.06 ± 7.021 | <0.001* |
Diastolic blood pressure (mm Hg) | 86.2 ± 3.94 | 87.31 ± 6.074 | 0.076 |
Hirsutism score | 5.53 ± 0.472 | 13.54 ± 2.51 | <0.001* |
Body mass index (kg/m2) | 30.28 ± 57 | 31.889.99 | 0.103 |
Waist/hip ratio | 1.11 ± 0.26 | 1.14 ± 0.30 | 0.392 |
FMI (kg/m2) | 7.66 ± 1.45 | 7.86 ± 1.579 | 0.260 |
FFMI (kg/m2) | 21.81 ± 4.13 | 22.42 ± 4.47 | 0.233 |
Ovarian volume | 4.33 ± 1.1 | 10.64 ± 1.3 | <0.001* |
AFC | 5.33 ± 1.1 | 12.14 ± 1.4 | <0.001* |
Total cholesterol (mg/dL) | 179.47 ± 19.7 | 198.8 ± 52.38 | <0.001* |
Triglycerides (mg/dL) | 150.23 ± 20.84 | 187.6 ± 62.14 | <0.001* |
LDL cholesterol (mg/dL) | 117.48 ± 19.05 | 165.98 ± 60.15 | <0.001* |
HDL cholesterol (mg/dL) | 56.58 ± 6.04 | 39.2 ± 13.63 | <0.001* |
Fasting plasma glucose (mg/dL) | 87.65 ± 9.14 | 108.2 ± 21.62 | <0.001* |
2-h plasma glucose (mg/dL) | 127.6 ± 24.2 | 163.67 ± 41.7 | <0.001* |
HbA1c (%) | 5.82 ± 0.182 | 6.35 ± 0.977 | <0.001* |
Fasting serum insulin (lU/mL) | 7.14 ± 1.427 | 20.05 ± 7.3 | <0.001* |
HOMA-IR | 1.55 ± 0.39 | 4.63 ± 1.87 | <0.001* |
HOMA-β | 169.7 ± 18.14 | 122.01 ± 43.6 | <0.001* |
FSH (mIU/mL) | 6.17 ± 1.53 | 6.6 ± 1.26 | <0.001* |
LH (mIU/mL) | 6.95 ± 0.646 | 10.3 ± 1.23 | <0.001* |
LH/FSH | 1.24 ± 0.54 | 1.6 ± 0.278 | <0.001* |
SHBG (nmol/L) | 53.58 ± 6.04 | 28.8 ± 4.71 | <0.001* |
DHEA-S (mg/mL) | 1.22 ± 0.33 | 2.37 ± 0.316 | <0.001* |
Androstenedione (ng/mL) | 1.41 ± 0.508 | 1.62 ± 0.557 | <0.001* |
Total testosterone (ng/mL) | 0.67 ± 0.18 | 1.1 ± 0.218 | <0.001* |
Free androgen index | 1.03 ± 0.36 | 2.56 ± 1.22 | <0.001* |
hs-CRP (μg/ml) | 2.61 ± 0.51 | 5.56 ± 0.98 | <0.001* |
WBC count (cell×103/μl) | 4.74 ± 0.397 | 6.97 ± 2.31 | <0.001* |
CIMT (mm) | 0.69 ± 0.06 | 1.18 ± 0.46 | <0.001* |
Serum ICAM-1 (ng/ml) | 87.8 ± 49.5 | 445.17 ± 184.95 | <0.001* |
Relative ICAM expression levels | 0.75 ± 0.48 | 4.57 ± 2.16 | <0.001* |
Clinical and laboratory characteristics of women with PCOS
Parameters | PCO patients (n = 180) | ||
---|---|---|---|
NGT Group (mean ± SD) (n = 75) | IGT Group (mean ± SD) (n = 65) | T2DM Group (mean ± SD) (n = 40) | |
Systolic blood pressure (mm Hg) | 127.76 ± 7.21 | 134.49 ± 4.74a | 139.67 ± 7.04b,c |
Diastolic blood pressure (mm Hg) | 83.8 ± 3.86 | 87.85 ± 6.33a | 90.98 ± 5.80b,c |
Hirsutism score | 13.78 ± 2.594 | 13.42 ± 2.9 | 13.29 ± 1.42 |
Body mass index (kg/m2) | 32.96 ± 14.25 | 30.11 ± 4.41 | 32.65 ± 5.73 |
Waist/hip ratio | 0.96 ± 0.212 | 1.34 ± 0.27a | 1.17 ± 0.27b,c |
Ovarian volume | 9.88 ± 0.491 | 10.75 ± 1.21a | 11.89 ± 1.4b,c |
AFC | 11.38 ± 0.491 | 12.25 ± 1.17a | 13.39 ± 1.6b,c |
FMI (kg/m2) | 6.20 ± 0.68 | 8.7 ± 0.63a | 9.49 ± 0.63b,c |
FFMI (kg/m2) | 17.65 ± 1.96 | 25.02 ± 1.82a | 27.17 ± 1.06b,c |
Total cholesterol (mg/dL) | 191.21 ± 54.43 | 201.5 ± 54.43 | 209.02 ± 43.32 |
Triglycerides (mg/dL) | 174.02 ± 55.3 | 184.2 ± 57.19 | 216.04 ± 73.1b,c |
LDL cholesterol (mg/dL) | 168.15 ± 61.11 | 163.17 ± 57.96 | 166.46 ± 63.11 |
HDL cholesterol (mg/dL) | 41.46 ± 11.78 | 36.21 ± 13.85a | 39.8 ± 15.8 |
FPG (mg/dL) | 91.1 ± 6.84 | 108.16 ± 12.50a | 140.5 ± 12.7b,c |
2-h blood glucose (mg/dL) | 130.28 ± 23.2 | 171.94 ± 28.68a | 212.8 ± 30.1b,c |
FSI (lU/mL) | 13.5 ± 3.289 | 26.96 ± 5.51a | 21.1 ± 3.34b,c |
HOMA-IR | 2.85 ± .855 | 6.25 ± 1.44a | 5.34 ± 0.647b,c |
HOMA-β | 139.23 ± 54.11 | 124.4 ± 35.3a | 108.6 ± 41.55b,c |
HbA1c (%) | 5.66 ± 0.35 | 6.37 ± 0.615a | 7.62 ± 0.983b,c |
Total testosterone (ng/mL) | 0.9 ± 0.231 | 0.99 ± 0.304a | 1.07 ± 0.155b,c |
Free androgen index | 1.813 ± 0.98 | 2.88 ± 0.81a | 3.4 ± 0.69b,c |
FSH (mIU/mL) | 6.02 ± 1.314 | 7.107 ± 1.06a | 7.05 ± 0.985b |
LH (mIU/mL) | 9.45 ± 0.491 | 10.44 ± 1.15a | 11.98 ± 0.201b,c |
LH/FSH | 1.63 ± 0.341 | 1.48 ± 0.144a | 1.73 ± 0.241c |
SHBG (nmol/L) | 33.54 ± 3.108 | 26.23 ± 2.011a | 24.197 ± 0.96b,c |
DHEA-S (mg/mL) | 2.17 ± 0.245 | 2.43 ± 0.241a | 2.66 ± 0.286b,c |
Androstenedione (ng/mL) | 1.23 ± 0.371 | 1.91 ± 0.539a | 1.88 ± 0.43 |
hs-CRP (μg/ml) | 4.92 ± 0.761 | 5.79 ± 0.760a | 6.37 ± 0.91b,c |
WBC count (cell×103/μl) | 4.79 ± 0.713 | 8.45 ± 2.27a | 8.58 ± 1.281b,c |
CIMT (mm) | 0.96 ± 0.263 | 1.09 ± 0.36a | 1.74 ± 0.467b,c |
Serum ICAM-1 (ng/ml) | 281.3 ± 95.29 | 509.4 ± 82.16a | 594.6 ± 156.12b,c |
Relative ICAM expression levels | 2.68 ± 0.85 | 5.7 ± 1.52a | 6.05 ± 2.119b,c |
Comparison of expression and serum levels of ICAM-1 in studied groups
Correlation between serum and expression levels of ICAM-1 with clinical and biochemical parameters of PCOS patients
Characteristics | Serum ICAM-1 levels | ICAM-1 expression levels | ||
---|---|---|---|---|
r | p | r | p | |
Hirsutism score | 0.095 | 0.205 | 0.095- | 0.210 |
Body mass index (kg/m2) | 0.086 | 0.253 | 0.073 | 0.332 |
Waist/hip ratio | 0.494 | <0.001* | 0.501 | <0.001* |
Ovarian volume | 0.187 | 0.012* | 0.180 | 0.016 |
AFC | 0.187 | <0.001* | 0.180 | <0.001* |
FMI% | 0.689 | <0.001* | 0.709 | <0.001* |
FFMI% | 0.677 | <0.001* | 0 .666 | <0.001* |
Total cholesterol (mg/dL) | 0.111 | 0.137 | 0.114 | 0.128 |
Triglycerides (mg/dL) | 0.019 | 0.797 | 0.030 | 0.694 |
LDL cholesterol (mg/dL) | 0.014 | 0.853 | 0.008 | 0.912 |
HDL cholesterol (mg/dL) | 0.036 | 0.631 | 0.044 | 0.561 |
FPG (mg/dL) | 0.435 | <0.001* | 0.477 | <0.001* |
2-h blood glucose (mg/dL) | 0.471 | <0.001* | 0.438 | <0.001* |
FSI (lU/mL) | 0.854 | <0.001* | 0.794 | <0.001* |
HOMA-IR | 0.874 | <0.001* | 0.932 | <0.001* |
HbA1c (%) | 0.404 | <0.001* | 0.369 | <0.001* |
HOMA-β | −0.147 | <0.001* | 0.222 | <0.001* |
Total testosterone (ng/mL) | 0.114 | <0.001* | 0.121 | <0.001* |
Free androgen index | 0.466 | <0.001* | 0.469 | <0.001* |
FSH (mIU/mL) | 0.502 | <0.001* | 0.518 | <0.001* |
LH (mIU/mL) | 0.549 | <0.001* | 0.561 | <0.001* |
SHBG (nmol/L) | −0.658 | <0.001* | 0.639 | <0.001* |
DHEA-S (mg/mL) | 0.269 | <0.001* | 0.262 | <0.001* |
Androstenedione (ng/mL) | 0.613 | <0.001* | 0.625 | <0.001* |
hs-CRP (μg/ml) | 0.342 | <0.001* | 0.374 | <0.001* |
WBC count (cell×103/μl) | 0.602 | <0.001* | 0.650 | <0.001* |
CIMT (mm) | 0.252 | <0.001* | 0.257 | <0.001* |
Linear regression analysis with serum ICAM-1 levels as dependent variable in PCOS groups
Model | Unstandardized Coefficients | Standardized Coefficients | t | P value | 95% C.I. | |||
---|---|---|---|---|---|---|---|---|
B | SE | Beta | Lower Bound | Upper Bound | ||||
1 | (Constant) | 135.851 | 80.832 | 1.681 | 0.095 | −23.727- | 295.428 | |
Total cholesterol | −0.019- | 0.103 | 0.006- | 0.190 | 0.850 | −0.222- | 0.183 | |
LDL cholesterol | −0.001- | 0.087 | 0.000 | 0.014 | 0.989 | −0.172- | 0.170 | |
FPG | −0.163- | 0.456 | 0.019 | 0.357 | 0.721 | −1.063- | 0.737 | |
HOMA-IR | 90.239 | 4.100 | 0.917 | 22.008 | < 0.001* | 82.144 | 98.334 | |
Body mass index | −0.215- | 0.537 | 0.012- | 0.400 | 0.689 | −1.274- | 0.844 | |
LH | 1.267 | 7.087 | 0.008 | 0.179 | 0.858 | −12.725- | 15.259 | |
hs-CRP | −4.453- | 7.328 | 0.024 | 0.608 | 0.544 | −18.919- | 10.014 | |
WBC count | 2.683 | 3.020 | 0.034 | 0.888 | 0.376 | −3.280- | 8.645 | |
Ovarian volume | −7.053- | 5.234 | 0.050 | 1.347 | 0.180 | −17.386- | 3.280 | |
Hirsutism score | −2.981- | 2.232 | 0.040 | 1.336 | 0.184 | −7.388- | 1.426 | |
CIMT | 23.455 | 15.458 | 0.059 | 1.517 | 0.131 | −7.061- | 53.972 |
Linear regression analysis with ICAM-1 expression levels as dependent variable in PCOS groups
Model | Unstandardized Coefficients | Standardized Coefficients | t | P value | 95% C.I. | |||
---|---|---|---|---|---|---|---|---|
B | SE | Beta | Lower Bound | Upper Bound | ||||
1 | (Constant) | −0.928 | 1.221 | 0.760 | 0.448 | −3.340 | 1.483 | |
Total cholesterol | −0.001 | 0.002 | 0.017 | 0.456 | 0.649 | −0.004 | 0.002 | |
LDL cholesterol | 0.000 | 0.001 | 0.012 | 0.340 | 0.734 | −0.002 | 0.003 | |
FPG | 0.004 | 0.007 | 0.036 | 0.526 | 0.599 | −0.010 | 0.017 | |
HOMA-IR | 0.829 | 0.062 | 0.720 | 13.389 | < 0.001* | 0.707 | 0.952 | |
Body mass index | 0.000 | 0.008 | 0.002 | 0.052 | 0.958 | −0.016 | 0.016 | |
LH | 0.114 | 0.107 | 0.065 | 1.069 | 0.287 | −0.097 | 0.326 | |
hs-CRP | 0.139 | 0.111 | 0.063 | 1.251 | 0.213 | −0.080 | 0.357 | |
WBC count | 0.152 | 0.046 | 0.162 | 3.328 | 0.001* | 0.062 | 0.242 | |
Ovarian volume | −0.178- | 0.079 | 0.0107 | 2.247 | 0.026* | −0.334 | −0.022 | |
Hirsutism score | 0.012 | 0.034 | 0.014 | 0.368 | 0.713 | −0.054 | 0.079 | |
CIMT | 0.039 | 0.234 | 0.008 | 0.168 | 0.867 | −0.422 | 0.500 |