The aim of this study is to assess the association of two polymorphisms, the cartilage
intermediate layer protein 2 (
Increased blood lipid and lipoprotein levels, low HDL cholesterol concentration, glucose intolerance, hypertension, and obesity have emerged as some of the most serious public health concerns in recent decades. These variables are closely related to a number of pathological disorders including cardiovascular disease (CVD). Although recent increases in CVD risk factors often reflect lifestyle changes, genetic factors also play a substantial role. Genome-wide association studies have revealed the association of DNA polymorphisms in both the
The
Angiotensin converting enzyme (
The aim of this study is to examine if genetic variants in both
This study is a part of cross-sectional survey conducted in Slovakia between 2009 and 2013 to analyze the effect of menopause on biomarkers of health in pre- and postmenopausal women. The entire study sample comprised 341 women ranging in age from 39 to 65 years (mean age =
Data concerning lifestyle habits including physical activity, smoking, and their health status and menstrual cycle characteristics were investigated by assisted questionnaire.
Women were divided according to their menopausal status (MS) into pre- and postmenopausal groups, in accordance with the WHO definition [
All anthropometrical parameters were measured by professional anthropologists and the same instruments were used for all subjects. Anthropometric measurements were taken using standard anthropometric technique [
Venous blood was collected after overnight fasting. The plasma was separated and biochemical analysis of gamma glutamyl transpeptidase (GMT), alanine aminotransferase (ALT), uric acid (UA), total cholesterol (TC), bilirubin (Bil), fasting blood glucose (Glc), total cholesterol (TC), TG, HDL-C, apolipoprotein A1 (apoA1), and apolipoprotein B (apoB) was carried out by routine laboratory methods in the Department of Clinical Laboratories of the Bratislava Alpha Medical. The LDL-C levels were calculated using the Friedewald formula [
Genomic DNA was extracted from peripheral blood samples using the SiMax Genomic DNA Extraction kit (Ecoli). PCR was used to detect the presence of the insertion (I) and deletion (D) alleles in intron 16 of the
The Chi-square test was used to analyze whether the genotypic distribution and the alleles were in accordance with the Hardy-Weinberg equilibrium. An analysis of covariance was used to determine differences in variables over the menopausal status,
The mean values of anthropometric, biochemical characteristics and blood pressure of women in the entire sample by menopausal status are shown in Table
Anthropometrical, biochemical variables and blood pressure in Slovak women by menopausal status.
Parameter | Total | Premenopause | Postmenopause |
|
---|---|---|---|---|
|
|
|
||
Age (years)* |
|
|
|
|
Weight (kg)* |
|
|
|
0.786 |
WC (cm) |
|
|
|
0.722 |
HC (cm)* |
|
|
|
0.804 |
BMI |
|
|
|
0.307 |
WHR |
|
|
|
0.602 |
GMT (µkat/L)* |
|
|
|
|
ALT (µkat/L)* |
|
|
|
|
UA (µmol/L) |
|
|
|
0.075 |
TC (mmol/L) |
|
|
|
0.840 |
TG (mmol/L)* |
|
|
|
0.621 |
| ||||
|
|
|
||
| ||||
Bilirubin (µmol/L)* |
|
|
|
0.831 |
Glucose (mmol/L)* |
|
|
|
0.103 |
| ||||
|
|
|
||
| ||||
HDL-C (mmol/L) |
|
|
|
0.141 |
LDL-C (mmol/L) |
|
|
|
0.734 |
apoA1 (g/L)* |
|
|
|
0.522 |
apoB (g/L) |
|
|
|
0.711 |
apoB-to-apoA1 |
|
|
|
0.537 |
TC-to-HDL-C* |
|
|
|
0.176 |
non-HDL-C |
|
|
|
0.624 |
LDL-C-to-HDL-C* |
|
|
|
0.171 |
|
|
|
|
0.239 |
| ||||
|
|
|
||
| ||||
sBP (mmHg)* |
|
|
|
0.581 |
dBP (mmHg)* |
|
|
|
0.508 |
WC: waist circumference; HC: hip circumference; BMI: body mass index; WHR: waist-to-hip ratio; GMT: gamma glutamyl transpeptidase, ALT: alanine aminotransferase; UA: uric acid; TC: total cholesterol; TG: triglycerides, HDL-C: high density lipoprotein cholesterol; LDL-C: low density lipoprotein; apoA1: apolipoprotein A1; apoB: apolipoprotein B; sBP: systolic blood pressure; dBP: diastolic blood pressure. Values represent mean ± SD.
The genotype distribution and allele frequencies of the
To address the association and impact of the
Anthropometrical, biochemical variables and blood pressure according to
Parameter |
|
Menopause status * |
||||
---|---|---|---|---|---|---|
GG | GT/TT |
|
|
|
| |
|
|
|||||
Weight (kg) |
|
|
0.303 | 0.583 | ||
WC (cm) |
|
|
0.479 | 0.489 | ||
HC (cm) |
|
|
0.012 | 0.911 | ||
BMI (kg/m2) |
|
|
1.203 | 0.274 | ||
WHR |
|
|
1.386 | 0.240 | ||
GMT (µkat/L) |
|
|
0.910 | 0.341 | ||
ALT (µkat/L) |
|
|
0.949 | 0.331 | ||
UA (µmol/L) |
|
|
0.000 | 0.996 | ||
TC (mmol/L) |
|
|
3.629 | 0.058 | ||
TG (mmol/L) |
|
|
2.733 | 0.099 | ||
| ||||||
|
|
|||||
| ||||||
Bilirubin (µmol/L) |
|
|
0.963 | 0.327 | ||
Glucose (mmol/L) |
|
|
1.032 | 0.310 | ||
| ||||||
|
|
|
||||
| ||||||
HDL-C (mmol/L) |
|
|
7.810 |
|
|
0.882 |
LDL-C (mmol/L) |
|
|
6.076 |
|
|
0.557 |
apoA1 (g/L) |
|
|
0.689 | 0.407 | ||
apoB (g/L) |
|
|
9.289 |
|
|
0.821 |
apoB-to-apoA1 |
|
|
10.572 |
|
|
0.500 |
TC-to-HDL-C |
|
|
8.601 |
|
|
0.304 |
non-HDL-C |
|
|
7.497 |
|
|
0.485 |
LDL-C-to-HDL-C |
|
|
8.454 |
|
|
0.352 |
|
|
|
2.989 | 0.085 | ||
| ||||||
|
|
|||||
| ||||||
sBP (mmHg) |
|
|
0.956 | 0.329 | ||
dBP (mmHg) |
|
|
0.052 | 0.820 |
WC: waist circumference; HC: hip circumference; BMI: body mass index; WHR: waist-to-hip ratio; GMT: gamma glutamyl transpeptidase, ALT: alanine aminotransferase; UA: uric acid; TC: total cholesterol; TG: triglycerides, HDL-C: high density lipoprotein cholesterol; LDL-C: low density lipoprotein; apoA1: apolipoprotein A1; apoB: apolipoprotein B; sBP: systolic blood pressure; dBP: diastolic blood pressure. Values represent mean ± SD.
Bonferonni pairwise comparisons between
Parameter |
|
Estimated marginal mean | SE | 95% CI | Mean difference | SE |
|
95% CI for difference | |||
---|---|---|---|---|---|---|---|---|---|---|---|
HDL-C | GT/TT | 1.74 | 0.07 | 1.60 | 1.88 | GT/TT versus GG | 0.20 | 0.07 |
|
0.06 | 0.35 |
GG | 1.54 | 0.02 | 1.49 | 1.58 | |||||||
LDL-C | GT/TT | 2.84 | 0.18 | 2.49 | 3.19 | GT/TT versus GG |
|
0.19 |
|
|
|
GG | 3.29 | 0.06 | 3.17 | 3.40 | |||||||
apoB | GT/TT | 0.81 | 0.05 | 0.72 | 0.90 | GT/TT versus GG |
|
0.05 |
|
|
|
GG | 0.95 | 0.01 | 0.92 | 0.98 | |||||||
apoB-to-apoA1 | GT/TT | 0.47 | 0.03 | 0.41 | 0.53 | GT/TT versus GG |
|
0.03 |
|
|
|
GG | 0.58 | 0.01 | 0.56 | 0.60 | |||||||
TC-to-HDL-C | GT/TT | 3.18 | 0.20 | 2.79 | 3.56 | GT/TT versus GG |
|
0.03 |
|
|
|
GG | 3.76 | 0.06 | 3.64 | 3.88 | |||||||
non-HDL-C | GT/TT | 3.39 | 0.19 | 3.01 | 3.77 | GT/TT versus GG |
|
0.20 |
|
|
|
GG | 3.92 | 0.06 | 3.80 | 4.04 | |||||||
LDL-C-to-HDL-C | GT/TT | 1.81 | 0.17 | 1.48 | 2.14 | GT/TT versus GG |
|
0.17 |
|
|
|
GG | 2.29 | 0.05 | 2.19 | 2.40 |
Based on estimated marginal means.
Table
Anthropometrical, biochemical variables and blood pressure according to
Parameter |
|
DD versus ID versus II | II versus ID/DD | Menopausal status * |
|||||
---|---|---|---|---|---|---|---|---|---|
DD | ID | II |
|
|
|
|
|
| |
|
|
|
|||||||
Weight (kg) |
|
|
|
0.062 | 0.940 | 0.024 | 0.877 | ||
WC (cm) |
|
|
|
0.697 | 0.499 | 1.018 | 0.314 | ||
HC (cm) |
|
|
|
0.549 | 0.578 | 0.347 | 0.556 | ||
BMI (kg/m2) |
|
|
|
0.021 | 0.979 | 0.027 | 0.868 | ||
WHR |
|
|
|
2.422 | 0.090 | 1.162 | 0.282 | ||
GMT (µkat/L) |
|
|
|
0.135 | 0.873 | 0.121 | 0.729 | ||
ALT (µkat/L) |
|
|
|
1.109 | 0.331 | 1.990 | 0.159 | ||
UA (µmol/L) |
|
|
|
0.096 | 0.908 | 0.031 | 0.860 | ||
TC (mmol/L) |
|
|
|
2.189 | 0.114 | 4.393 |
|
|
0.377 |
TG (mmol/L) |
|
|
|
1.091 | 0.337 | 0.008 | 0.927 | ||
| |||||||||
|
|
|
|||||||
| |||||||||
Bilirubin (µmol/L) |
|
|
|
1.354 | 0.260 | 0.712 | 0.399 | ||
| |||||||||
|
|
|
|||||||
| |||||||||
Glucose (mmol/L) |
|
|
|
0.070 | 0.933 | 0.008 | 0.928 | ||
| |||||||||
|
|
|
|||||||
| |||||||||
HDL-C (mmol/L) |
|
|
|
0.393 | 0.676 | 0.480 | 0.489 | ||
LDL-C (mmol/L) |
|
|
|
2.619 | 0.075 | 5.011 |
|
|
0.975 |
apoA1 (g/L) |
|
|
|
1.855 | 0.158 | 3.646 | 0.057 | ||
apoB (g/L) |
|
|
|
1.603 | 0.203 | 3.172 | 0.076 | ||
apoB-to-apoA1 |
|
|
|
0.019 | 0.981 | 0.035 | 0.851 | ||
TC-to-HDL-C |
|
|
|
0.548 | 0.579 | 0.981 | 0.323 | ||
non-HDL-C |
|
|
|
2.268 | 0.105 | 4.549 |
|
|
0.951 |
LDL-C-to-HDL-C |
|
|
|
0.660 | 0.517 | 1.319 | 0.252 | ||
|
|
|
|
0.492 | 0.612 | 0.004 | 0.949 | ||
| |||||||||
|
|
|
|||||||
| |||||||||
sBP (mmHg) |
|
|
|
0.781 | 0.459 | 1.509 | 0.220 | ||
dBP (mmHg) |
|
|
|
0.260 | 0.771 | 0.451 | 0.503 |
WC: waist circumference; HC: hip circumference; BMI: body mass index; WHR: waist-to-hip ratio; GMT: gamma glutamyl transpeptidase, ALT: alanine aminotransferase; UA: uric acid; TC: total cholesterol; TG: triglycerides, HDL-C: high density lipoprotein cholesterol; LDL-C: low density lipoprotein; apoA1: apolipoprotein A1; apoB: apolipoprotein B; sBP: systolic blood pressure; dBP: diastolic blood pressure. Values represent mean ± SD. aAdjusted for age; badjusted for age, BMI, and WHR.
Bonferonni pairwise comparisons between
Parameter |
|
Estimated marginal mean | SE | 95% CI | Mean difference | SE |
|
95% CI for difference | |||
---|---|---|---|---|---|---|---|---|---|---|---|
LDL-C | II | 3.02 | 0.11 | 2.79 | 3.24 | II versus ID/DD | −0.29 | 0.13 |
|
|
|
DD/ID | 3.31 | 0.06 | 3.18 | 3.43 | |||||||
TC | II | 5.21 | 0.12 | 4.98 | 5.45 | II versus ID/DD | −0.29 | 0.14 |
|
|
|
DD/ID | 5.50 | 0.07 | 5.37 | 5.63 | |||||||
non-HDL-C | II | 3.62 | 0.12 | 3.38 | 3.87 | II versus ID/DD | −0.33 | 0.14 |
|
|
|
DD/ID | 3.95 | 0.07 | 3.81 | 4.08 |
Based on estimated marginal means.
Adjustment for multiple comparisons: Bonferroni.
A stepwise regression analysis was used to test the independent impact of
Regression analysis of selected confounder effects on lipid, and lipoprotein levels, and atherogenic indices in Slovak women.
Dependent variable | Independent variable |
|
SE | Beta |
|
95.0% CI for |
Collinearity statistics tolerance | |
---|---|---|---|---|---|---|---|---|
TC | ln age | 1.72 | 0.52 | 0.19 | 0.001 | 0.70 | 2.73 | 0.99 |
|
Current smoker | 0.32 | 0.09 | 0.20 | 0.001 | 0.14 | 0.50 | 0.98 |
|
0.40 | 0.14 | 0.16 | 0.005 | 0.12 | 0.69 | 0.98 | |
|
||||||||
Excluded variables: menopausal status, BMI, WHR, former sport activities, and recent sport activities, current smoker. | ||||||||
LDL-C | ln age | 1.25 | 0.51 | 0.15 | 0.016 | 0.24 | 2.26 | 0.99 |
|
Current smoker | 0.29 | 0.09 | 0.19 | 0.002 | 0.11 | 0.47 | 0.98 |
|
0.37 | 0.14 | 0.16 | 0.009 | 0.10 | 0.65 | 0.98 | |
|
0.42 | 0.20 | 0.13 | 0.035 | 0.03 | 0.81 | 1.00 | |
|
||||||||
Excluded variables: menopausal status, BMI, WHR, former sport activities, and recent sport activities. | ||||||||
non-HDL-C | ln age | 1.52 | 0.59 | 0.16 | 0.010 | 0.37 | 2.67 | 0.91 |
|
WHR | 2.39 | 0.84 | 0.17 | 0.005 | 0.73 | 4.05 | 0.91 |
Current smoker | 0.33 | 0.10 | 0.20 | 0.001 | 0.14 | 0.53 | 0.98 | |
|
0.41 | 0.16 | 0.16 | 0.008 | 0.11 | 0.72 | 0.97 | |
|
0.52 | 0.22 | 0.14 | 0.018 | 0.09 | 0.94 | 1.00 | |
|
||||||||
Excluded variables: menopausal status, BMI, former sport activities, and recent sport activities. | ||||||||
HDL-C | ln age | 0.49 | 0.20 | 0.14 | 0.017 | 0.09 | 0.89 | 0.92 |
|
ln BMI | −0.59 | 0.14 | −0.28 | <0.001 | −0.86 | −0.32 | 0.69 |
WHR | −0.89 | 0.35 | −0.17 | 0.010 | −1.57 | −0.21 | 0.67 | |
|
−0.21 | 0.08 | −0.14 | 0.009 | −0.36 | −0.05 | 0.99 | |
|
||||||||
Excluded variables: menopausal status, former sport activities, recent sport activities, and current smoker. | ||||||||
apoB | ln BMI | 0.26 | 0.08 | 0.19 | 0.001 | 0.10 | 0.41 | 0.99 |
|
Current smoker | 0.05 | 0.02 | 0.12 | 0.038 | 0.00 | 0.09 | 1.00 |
|
0.14 | 0.05 | 0.15 | 0.011 | 0.03 | 0.24 | 1.00 | |
|
||||||||
Excluded variables: age, menopausal status, WHR, former sport activities, and recent sport activities. | ||||||||
apoB-to-apoA1 | ln BMI | 0.23 | 0.05 | 0.25 | <0.001 | 0.13 | 0.34 | 0.99 |
|
Current smoker | 0.04 | 0.02 | 0.13 | 0.026 | 0.00 | 0.07 | 1.00 |
|
0.11 | 0.04 | 0.16 | 0.004 | 0.03 | 0.18 | 1.00 | |
|
||||||||
Excluded variables: age, menopausal status, WHR, former sport activities, and recent sport activities. | ||||||||
ln TC-to-HDL-C | ln BMI | 0.37 | 0.10 | 0.24 | <0.001 | 0.17 | 0.57 | 0.70 |
|
WHR | 0.71 | 0.25 | 0.18 | 0.005 | 0.21 | 1.20 | 0.70 |
Current smoker | 0.06 | 0.03 | 0.13 | 0.022 | 0.01 | 0.11 | 1.00 | |
|
0.17 | 0.06 | 0.16 | 0.004 | 0.06 | 0.29 | 1.00 | |
|
||||||||
Excluded variables: age, menopausal status, former sport activities, and recent sport activities. | ||||||||
ln LDL-C-to-HDL-C | ln BMI | 0.61 | 0.12 | 0.28 | <0.001 | 0.37 | 0.85 | 0.99 |
|
Current smoker | 0.08 | 0.04 | 0.13 | 0.024 | 0.01 | 0.15 | 1.00 |
|
0.25 | 0.08 | 0.17 | 0.003 | 0.08 | 0.42 | 1.00 | |
|
||||||||
Excluded variables: age, menopausal status, WHR, former sport activities, and recent sport activities. |
Table
The common impact of
Dependent variable | ||||||
---|---|---|---|---|---|---|
Factor | LDL-C | non-HDL-C | ||||
|
|
|
Mean | SD | Mean | SD |
II | GT/TT | 7 |
|
0.74 |
|
0.74 |
GG | 56 | 3.09 | 0.82 | 3.73 | 0.98 | |
ID/DD | GT/TT | 21 | 2.95 | 0.70 | 3.51 | 0.81 |
GG | 197 |
|
0.98 |
|
1.09 | |
|
|
|
The association between the
Association between
Association between
In this study, we determined a profound impact of the
When evaluating the impact of
We have considered common influence of menopausal status and
In addition, the findings in this study showed a lack of association between the
In accordance with other studies [
The results of this study indicate that the minor T allele of the
The authors wish to thank Dr. Pavel Blažíček for biochemical analysis and Dr. Ladislava Wsólová for statistical advice. This study was supported by the Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic (VEGA 1/0247/09, 1/0493/13).