Figure 4 and 5 show the results from the genotyping of 12 genetic variants in risk genes in diabetic patients with and without CVD.
1.1. Results from the genotyping of Factor V Leiden and HR2
Altogether for all patients a frequency of 5,5% is found – more than two-fold increase than the world population frequency of 1,9% and 2,9% in Europe. According to 1000 Genomes database the frequency of the heterozygotes is 2% and in our cohort we found it 11%. No connection between the mutation of FV Leiden and cardiovascular complications has been established, even in the group of the patients without CVD a higher frequency of the mutation is found – 9,4%. We found also a higher than population frequency for FV H1299R (R2) – 9,7% in comparison to world frequency 5,7% and 6% in Europe.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
FV (Leiden)
|
2.5%
|
9.4%
|
5.5%
|
FV G/A
|
5%
|
18.8%
|
11%
|
FV A/A
|
0
|
0
|
0
|
FV (R2)
|
8.8%
|
10.7%
|
9.7%
|
FV H/R
|
17.6%
|
21.4%
|
19.4%
|
FV R/R
|
0
|
0
|
0
|
Table 2 – Allelic and genotypic frequencies of FV G1691A (Leiden) and FV H1299R (R2)
1.2. Results from the genotyping of Prothrombin
We found a frequency of 1,4% and it is comparable to the world population frequency of 0,8% and to that in Europe – 1,1%.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
Prothrombin 20210A
|
2.5%
|
0
|
1.4%
|
Prothrombin G/A
|
5%
|
0
|
2.8%
|
Prothrombin A/A
|
0
|
0
|
0
|
Table 3 - Allelic and genotypic frequencies of Prothrombin G20210A
1.3. Results from the genotyping of PAI – 1
We found higher frequency of the pathogenic allele – 58,6% compared to 26,9% world population frequency according to Ensambl and 38,9% in Europe. The frequency of the homozygotes is 31,4% in comparison to 20,9% in the world and 29,4% in Europe.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
PAI-1 4G
|
65%
|
50%
|
58.6%
|
PAI-1 4G/5G
|
60%
|
46.7%
|
54.3%
|
PAI-1 4G/4G
|
35%
|
26.7%
|
31.4%
|
Table 4 - Allelic and genotypic frequencies of PAI-1 4G/5G
1.4. Results from the genotyping of Factor XIII
In our cohort was found lower frequency of the minor allele of 11,1% compared to 21,9% world population frequency and 25,2% in Europe. It is important to note that in the group with CVD the frequency is even lower – 7,5%, which suggests a protective role of that genetic variant.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
Factor XIII 34L
|
7.5%
|
15.6%
|
11.1%
|
Factor XIII V/L
|
15%
|
31.2%
|
22.2%
|
Factor XIII L/L
|
0
|
0
|
0
|
Table 5 - Allelic and genotypic frequencies of Factor XIII V34L
1.5. Results from the genotyping of β-Fibrinogen
The allelic frequency in our group is 22,2% which is higher than the world population frequency – 16,9%, and that in Europe – 20,3%. According to 1000 Genomes database the population frequency of the heterozygotes is 22% and we found it 38,9%. It increases statistically significant in the group with CVD compared to the one without CVD – 55% versus 18,7% - figure 11.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
β-Fibrinogen - 455 A
|
27.5%
|
15.6%
|
22.2%
|
β-Fibrinogen - 455 G/A
|
55%
p<0.03
|
18.7%
|
38.9%
|
β-Fibrinogen -455A/A
|
0
|
6.2%
|
2.7%
|
Table 6 - Allelic and genotypic frequencies of β-Fibrinogen -455 G/A
1.6. Results from the genotyping of HPA1
The found from us allelic frequency is 12,5% and is comparable to the world population frequency – 12,1%; in Europe – 15,2%.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
HPA-1b
|
7.5%
|
18.7%
|
12.5%
|
HPA-1a/1b
|
15%
|
37.4%
|
25%
|
HPA-1b/1b
|
0
|
0
|
0
|
Table 7 - Allelic and genotypic frequencies of GPIIIa L33P (HPA-1)
In order to conclude about the factors contributing to congenital thrombophilia we found higher frequencies for most of them than in the world population frequency but not reaching statistical significance. The highest frequency is that of the PAI-1 variant in patients with DM. The frequency of Factor XIII polymorphism is lower than that in world population frequency which is in accordance to the suggested protective role of the polymorphism. When comparing the frequencies in the groups with and without CVD only the variants of PAI-1 and Fibrinogen show higher frequency in the group with CVD – figure 13.
1.7. Results from the genotyping of MTHFR
The allelic frequency of MTHFR 677T we found is 25% and is a little lower than that of world population – 31%, and in Europe – 32%. The allelic frequency of MTHFR 1298C in our study is 38,9% and is higher than that in world – 29%, and in Europe – 32%.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
MTHFR 677T
|
22.5%
|
28.1%
|
25%
|
MTHFR C/T
|
25%
|
31.2%
|
27.8%
|
MTHFR T/T
|
10%
|
12.5%
|
11.1%
|
MTHFR 1298C
|
47.5%
|
28.1%
|
38.9%
|
MTHFR A/C
|
55%
|
31.2%
|
44.4%
|
MTHFR C/C
|
20%
|
12.5%
|
16.7%
|
Table 8 - Allelic and genotypic frequencies of MTHFR C677T и MTHFR A1298C
1.8. Results from the genotyping of ACE
We found a frequency of the homozygotes of the pathologic allele that is 36,1% and is higher than population frequency in Europe – 25%.
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
ACE Del
|
55%
|
65.6%
|
59.7%
|
ACE I/D
|
50%
|
43.7%
|
47.2%
|
ACE D/D
|
30%
|
43.7%
|
36.1%
|
Table 9 - Allelic and genotypic frequencies of ACE I/D
1.9. Results from the genotyping of ApoB
The mutation was not found in any of the patients and its world population frequency is 1:5000.
1.10. Results from the genotyping of ApoE
The frequency of the risk allele E4 we found is 13,9% and is comparable to the world population frequency – 13,8%q and that in Europe – 16,1%
Allele/Genotype
|
DM with CVD
|
DM without CVD
|
All
|
Apo E3/E4
|
10%
|
18.7%
|
13.9%
|
Apo E2/E4
|
5%
|
0
|
2.8%
|
Apo E3/E3
|
85%
|
81.3%
|
83.3%
|
Table 10 - Allelic and genotypic frequencies of Apo E2/E3/E4