Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants
Aim. To evaluate a possible involvement of the F2, F5, MTHFR gene variants into ischemic stroke pathogenesis in population of Ukraine. Methods. Polymorphic variants were analyzed in unrelated 183 stroke patients, 100 individuals from the general population of Ukraine and 88 healthy individuals elder...
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Інститут молекулярної біології і генетики НАН України
2010
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| Цитувати: | Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants / P.F. Tatarskyy, A.M. Kucherenko, S.A. Kravchenko, D.V. Shulzhenko, S.M. Kuznetsova, L.A. Livshits // Вiopolymers and Cell. — 2010. — Т. 26, № 4. — С. 299-305. — Бібліогр.: 30 назв. — англ. |
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nasplib_isofts_kiev_ua-123456789-1541362025-02-09T20:44:59Z Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants Ішемічний інсульт серед населення України: можливе залучення поліморфних варіантів генів F2 G20210A, F5 G1691A та MTHFR C677T Ишемический инсульт среди населения Украины: возможное вовлечение полиморфных вариантов генов F2 G20210A, F5 G1691A и MTHFR C677T Tatarskyy, P.F. Kucherenko, A.M. Kravchenko, S.A. Shulzhenko, D.V. Kuznetsova, S.M. Livshits, L.A. Biomedicine Aim. To evaluate a possible involvement of the F2, F5, MTHFR gene variants into ischemic stroke pathogenesis in population of Ukraine. Methods. Polymorphic variants were analyzed in unrelated 183 stroke patients, 100 individuals from the general population of Ukraine and 88 healthy individuals elder than 65 years using PCR followed by RFLP analysis. Results. Unfavourable polymorphic variants F2 20210A, F5 1691A and MTHFR 677T were observed more frequently in patients with ischemic stroke comparing to control groups. Conclusions. F5 1691A and MTHFR 677T polymorphic variants are associated with the occurrence of ischemic stroke in women. F2 20210A is associated with the occurrence of ischemic stroke in men. Cumulative risk factor for stroke development is revealed in a combination of unfavorable polymorphic variants 20210A, 1691A and 677T of F2, F5 and MTHFR genes. Мета. Провести аналіз зв’язку поліморфнних варіантів генів F2, F5, MTHFR та патогенезу інсульту серед населення України. Методи. Алельні варіанти визначали серед 183 неспоріднених пацієнтів з інсультом, 100 індивідуумів із загальної популяції та 88 – здорових індивідуумів старших 65 років, методом ПЛР та ПДРФ-аналізу. Результати. «Несприятливі» поліморфні варіанти F2 20210A, F5 1691A та MTHFR 677T спостерігалися частіше серед пацієнтів з ішемічним інсультом порівняно з контрольними групами. Висновки. Поліморфні варіанти F5 1691A та MTHFR 677Т асоційовані з випадками ішемічного інсульту у жінок, варіант F2 20210A – з випадками ішемічного інсульту у чоловіків. Комбінація виявлених мутантних варіантів 1691А, 20210А і 677T генів F5, F2 і MTHFR проявляється як адитивний фактор ризику виникнення инсульту. Цель. Провести анализ ассоциации полиморфных вариантов генов F2, F5, MTHFR и патогенеза инсульта среди населения Украины. Методы. Аллельные варианты анализировали среди 183 неродственных пациентов с инсультом, 100 индивидуумов из общей популяции Украины и 88 здоровых индивидуумов старше 65 лет методами ПЦР и ПДРФ-анализа. Результаты. «Неблагоприятные» полиморфные варианты F2 20210A, F5 1691A и MTHFR 677T выявлялись чаще у пациентов с ишемическим инсультом по сравнению с контрольными группами. Выводы. Полиморфные варианты F5 1691A и MTHFR 677Т ассоциированы со случаями ишемического инсульта у женщин, вариант F2 20210A – со случаями ишемического инсульта у мужчин. Комбинации выявленных мутантных вариантов 1691А, 20210А и 677T генов F5, F2 и MTHFR проявляются как аддитивный фактор риска возникновения инсульта. 2010 Article Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants / P.F. Tatarskyy, A.M. Kucherenko, S.A. Kravchenko, D.V. Shulzhenko, S.M. Kuznetsova, L.A. Livshits // Вiopolymers and Cell. — 2010. — Т. 26, № 4. — С. 299-305. — Бібліогр.: 30 назв. — англ. 0233-7657 DOI: http://dx.doi.org/10.7124/bc.000163 https://nasplib.isofts.kiev.ua/handle/123456789/154136 575.11 + 577.21 en Вiopolymers and Cell application/pdf Інститут молекулярної біології і генетики НАН України |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine |
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English |
| topic |
Biomedicine Biomedicine |
| spellingShingle |
Biomedicine Biomedicine Tatarskyy, P.F. Kucherenko, A.M. Kravchenko, S.A. Shulzhenko, D.V. Kuznetsova, S.M. Livshits, L.A. Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants Вiopolymers and Cell |
| description |
Aim. To evaluate a possible involvement of the F2, F5, MTHFR gene variants into ischemic stroke pathogenesis in population of Ukraine. Methods. Polymorphic variants were analyzed in unrelated 183 stroke patients, 100 individuals from the general population of Ukraine and 88 healthy individuals elder than 65 years using PCR followed by RFLP analysis. Results. Unfavourable polymorphic variants F2 20210A, F5 1691A and MTHFR 677T were observed more frequently in patients with ischemic stroke comparing to control groups. Conclusions. F5 1691A and MTHFR 677T polymorphic variants are associated with the occurrence of ischemic stroke in women. F2 20210A is associated with the occurrence of ischemic stroke in men. Cumulative risk factor for stroke development is revealed in a combination of unfavorable polymorphic variants 20210A, 1691A and 677T of F2, F5 and MTHFR genes. |
| format |
Article |
| author |
Tatarskyy, P.F. Kucherenko, A.M. Kravchenko, S.A. Shulzhenko, D.V. Kuznetsova, S.M. Livshits, L.A. |
| author_facet |
Tatarskyy, P.F. Kucherenko, A.M. Kravchenko, S.A. Shulzhenko, D.V. Kuznetsova, S.M. Livshits, L.A. |
| author_sort |
Tatarskyy, P.F. |
| title |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants |
| title_short |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants |
| title_full |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants |
| title_fullStr |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants |
| title_full_unstemmed |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants |
| title_sort |
ischemic stroke in ukrainian population: possible involvement of the f2 g20210a, f5 g1691a and mthfr c677t gene variants |
| publisher |
Інститут молекулярної біології і генетики НАН України |
| publishDate |
2010 |
| topic_facet |
Biomedicine |
| url |
https://nasplib.isofts.kiev.ua/handle/123456789/154136 |
| citation_txt |
Ischemic stroke in Ukrainian population: possible involvement of the F2 G20210A, F5 G1691A and MTHFR C677T gene variants / P.F. Tatarskyy, A.M. Kucherenko, S.A. Kravchenko, D.V. Shulzhenko, S.M. Kuznetsova, L.A. Livshits // Вiopolymers and Cell. — 2010. — Т. 26, № 4. — С. 299-305. — Бібліогр.: 30 назв. — англ. |
| series |
Вiopolymers and Cell |
| work_keys_str_mv |
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2025-11-30T14:49:31Z |
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2025-11-30T14:49:31Z |
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1850227214315945984 |
| fulltext |
BIOMEDICINE
Ischemic stroke in Ukrainian population: possible
involvement of the F2 G20210A, F5 G1691A and
MTHFR C677T gene variants
P. F. Tatarskyy1, 3, A. M. Kucherenko 1, 3, S. A. Kravchenko1,
D. V. Shulzenko2, S. M. Kuznetsova2, L. A. Livshits1
1Institute of Molecular Biology and Genetics National Academy of Sciences of Ukraine
150, Akademika Zabolotnogo Str., Kyiv, Ukraine, 03680
2Gerontology Institute of the Academy of Medical Sciences of Ukraine
67, Vyshgorodska Str., Kyiv, Ukraine, 04114
3National Taras Shevchenko University of Kyiv
64, Volodymyrska Str., Kyiv, Ukraine, 01033
tatarskyy@yahoo.com
Aim. To evaluate a possible involvement of the F2, F5, MTHFR gene variants into ischemic stroke patho-
genesis in population of Ukraine. Methods. Polymorphic variants were analyzed in unrelated 183 stroke
patients, 100 individuals from the general population of Ukraine and 88 healthy individuals elder than 65
years using PCR followed by RFLP analysis. Results. Unfavourable polymorphic variants F2 20210A, F5
1691A and MTHFR 677T were observed more frequently in patients with ischemic stroke comparing to con-
trol groups. Conclusions. F5 1691A and MTHFR 677T polymorphic variants are associated with the oc-
currence of ischemic stroke in women. F2 20210A is associated with the occurrence of ischemic stroke in
men. Cumulative risk factor for stroke development is revealed in a combination of unfavorable polymor-
phic variants 20210A, 1691A and 677T of F2, F5 and MTHFR genes.
Keywords: Factor V Leiden (F5 G1691A) gene, prothrombin (F2 G20210A) gene, methylenetetrahydrofo-
late reductase (MTHFR C677T) gene, ischemic stroke, combined genotype.
Introduction. Nowadays, stroke is one of the main
causes of invalidization and mortality in the developed
countries, being a crucial medical and social problem
[1–8].
Stroke is defined as rapidly developing clinical
signs of focal (or global) disturbance of cerebral func-
tion with symptoms, lasting 24 h or longer or leading
to death, with no apparent cause other than of vascular
origin [3]. Stroke is considered to be a complex poly-
genic disorder arising from a wide number of gene–
gene and gene–environment interactions. Since the ad-
vent of molecular genetics in medicine it has been a fo-
cus of interest to clarify a role of mutations in various
candidate genes and their impact on stroke develop-
ment. A series of studies have been carried out to ex-
plain possible mechanisms of this ischemic event. The
results of the study are not entirely consistent, but the
majority of the data support the concept that thrombo-
sis is important for ischemic stroke development [1–6].
The factor V (F5 G1691A) Leiden and G20210A
variant of the prothrombin (F2) gene are clotting fac-
tor mutations that are associated with an increased
299
ISSN 0233-7657. Biopolymers and Cell. 2010. Vol. 26. N 4
Ó Institute of Molecular Biology and Genetics NAS of Ukraine, 2010
tendency toward venous thrombosis [1, 4]. The eviden-
ce of a role for these gene variants in the risk of ische-
mic stroke is controversial [1, 4, 6].
The G to A transition at nucleotide position 20210
in the 3'-untranslated region of F2 gene plays a regu-
latory role in the gene expression [1, 5, 6, 9]. Pro-
thrombin has procoagulant, anticoagulant and antifib-
rinolytic activities, and thus a disorder involving pro-
thrombin results in hemostasis multiple imbalances [1,
5]. The F2 G20210A gene mutation is associated with
an elevated risk of deep vein thrombosis and con-
sequently ischemic stroke [1, 6].
The point mutation of factor V (F5 G1691A) leads
to the structural change in the factor V molecule and
makes it less susceptible to inactivation by activated
protein C (APC) [7], resulting in a hypercoagulable
state when more factor Va is available within the pro-
thrombinase complex, thereby increasing the genera-
tion of thrombin leading to thrombosis and, therefore,
ischemic stroke [1, 4, 6–8].
Hyperhomoñysteinemia has been documented as
an independent risk factor for stroke [1, 4]. The most
common form of genetic hyperhomocysteinemia re-
sults from production of a thermolabile variant of me-
thylenetetrahydrofolate reductase with reduced en-
zymatic activity caused by C677T substitution in the
MTHFR gene [1, 4]. Moderate elevation of homocys-
teine in the plasma is an established risk factor in the
development of ischemic stroke and venous throm-
bosis due to its toxic influence on the endothelium.
Concerning the connection of C677T MTHFR and a
risk of stroke, the results are conflicting as well [1, 4].
The aim of this study was to evaluate a possible
involvement of the F2 G20210A, F5 G1691A, MTHFR
C677T gene variants into stroke pathogenesis.
Materials and methods. Study subjects. Unrelated
individuals who were randomly selected between 2008
and 2009 from different regions of Ukraine were inclu-
ded in this study. An informed consent was obtained
from each participant prior to blood collection and
DNA extraction.
This study was approved by the Ethical Committee
of the Institute of Molecular Biology and Genetics of
the National Academy of Sciences of Ukraine and by
state institution «Institute of Gerontology of the Acade-
my of Medical Sciences of Ukraine».
A case group included 183 patients (men – 95, wo-
men – 88), who have survived after ischemic stroke and
were referred for rehabilitation. The patients were do-
cumented according to a standardized protocol com-
prising the following information: vascular risk factors
including hypertension, diabetes, cigarette smoking,
body mass index, medical history including general
medical diseases (e. g. previous cardiac, neurological,
oncological diseases) and previous medication with
particular reference to vascular diseases (TIA/stroke,
venous thrombosis); laboratory and instrumental in-
vestigations including carotid and vertebral ultraso-
und, electroencephalography, echocardiography, MRI
or CT; stroke cause and stroke severity as measured by
validated scales. A control group I consisted of 100 in-
dividuals from the general population of Ukraine
(men – 50, women – 50). A control group II included
88 healthy individuals elder then 65 years (men – 35,
women – 53) without any history of ischemic stroke.
No significant difference between the age of men and
women within each group was observed.
Genotyping. DNA was extracted from the periphe-
ral blood leukocytes by standard phenol-chloroform
extraction method [10]. The F2 gene G20210A and F5
gene G1691A allelic variants were identified by multi-
plex polymerase chain reaction (PCR) with the use of
specific oligonucleotide primers followed by MnlI rest-
riction enzyme digestion, as described by Koksal et al.,
2006 [11].
The MTHFR gene C677T allele was detected by
PCR with the use of specific oligonucleotide primers
followed by HinfI restriction enzyme digestion, as des-
cribed by Michael et al., 2008 [12].
Electrophoresis of the PCR products was perfor-
med in a 1.8 % agarose gel. PCR products 169, 221 and
294 bp long correspondently were digested and run on
7 % PAGE, the gels were stained with EtBr followed
by visualization under UV.
Statistical analysis. The statistical analyses were
performed using MDR, Genepop and Arlequin soft-
wares [13–15]. Differences were considered signifi-
cant at P < 0.05 value of Fisher exact test.
Results and discussion. Because we included
individuals who have survived after ischemic stroke
only, one could argue that this could have led to pre-
judiced results. Case fatality in stroke largely depends
300
TATARSKYY P. F. ET AL.
on age, 88 % of all deaths from stroke occurring in
people older than 65 years [4].
The average age between the groups was signi-
ficantly different. In the control group II the individuals
average age (73.9 ± 6.4) was higher than in the case
group (64.6 ± 9.1), on the other hand, the average age
of individuals in the control group I (30.2 ± 7.6) was
lower than in the other groups.
To clarify a possible involvement of the F2
G20210A, F5 G1691A, MTHFR C677T in stroke the
analysis of above mentioned SNPs was performed.
Genotypes and allele frequencies of the three polymor-
phisms are presented in Table 1.
After the molecular-genetical analysis we have es-
tablished that the F2 20210A carriers frequency in the
case group was 4.4 %, and in the control group II such
301
ISCHEMIC STROKE IN UKRAINIAN POPULATION
Group
Loci
Control group
I n = 100
Men
n = 50
Women
n = 50
Control group
I² n = 88
Men
n = 35
Women
n = 53
Case group
n = 183
Men
n = 95
Women
n = 88
F5 G1691A
Genotype, n (%)
GG 98 (98) 50 (100) 48 (96) 85 (96.6) 35 (100) 50 (94.3) 177 (96.7) 94 (98.9) 83 (94.3)
GA 2 (2) 0 (0) 2 (4) 2 (2.3) 0 (0) 2 (3.8) 6 (3.3) 1 (1.1) 5 (5.7)
AA 0 (0) 0 (0) 0 (0) 1 (1.1) 0 (0) 1 (1.9) 0 (0) 0 (0) 0 (0)
Allele, n (%)
G 198 (99) 100 (100) 98 (98) 172 (97.7) 70 (100) 102 (96.2) 360 (98.4) 189 (99.5) 171 (97.2)
À 2 (1) 0 (0) 2 (2) 4 (2.3) 0 (0) 4 (3.8) 6 (1.6) 1 (0.5) 5(2.8)
F2 G20210A
Genotype, n (%)
GG 97 (97) 48 (96) 49 (98) 88 (100) 35 (100) 53 (100) 175 (95.6) 88 (92.6) 87 (98.9)
GA 3 (3) 2 (4) 1 (2) 0 (0) 0 (0) 0 (0) 8 (4.4)** 7 (7.4) 1 (1.1)
AA 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0) 0 (0)
Allele, n (%)
G 197 (98.5) 98 (98) 99 (99) 176 (100.0) 70 (100) 106 (100) 358 (97.8) 183 (96.3) 175 (99.4)
À 3 (1.5) 2 (2) 1 (1) 0 (0.0) 0 (0) 0 (0) 8 (2.2) 7 (3.7) 1 (0.6)
MTHFR C677T
Genotype, n (%)
CC 50 (50) 23 (46) 27 (54) 44 (50) 13 (37.1) 31 (58.5) 73 (39.9) 33 (34.7) 40 (45.5)
CT 45 (45) 25 (50) 20 (40) 37 (42) 20 (57.2) 17 (32.1) 87 (47.5) 49 (51.6) 38 (43.1)
TT 5 (5) 2 (4) 3 (6) 7 (8) 2 (5.7) 5 (9.4) 23 (12.6)* 13 (13.7) 10 (11.4)
CT+ TT 50 (50) 27 (54) 23 (46) 44 (50) 22 (62.9) 22 (41.5) 110 (60.1) 62 (65.3) 48(54.5)**
Allele, n (%)
C 145 (72.5) 71 (71) 74 (74) 125 (71.0) 46 (65.7) 79 (74.5) 233 (63.7) 115 (60.5) 118 (67)
T 55 (27.5) 29 (29) 26 (26) 51 (29.0) 24 (34.3) 27 (25.5) 133 (36.3) 75 (39.5) 58 (33)
n – group size; F2: A – 20210A; G – 20210G, F5: A – 1691A; G – 1691G, MTHFR: T – 677T, C – 677C, **Significant at P < 0.05 between
case and control group II, *Significant at P < 0.05 between case and control group I.
Table 1
Genotype and allele distribution among the study groups
carriers were not found. Significant difference was fo-
und between the frequency of men and women carriers
of the F2 20210A from the case group, men had higher
frequency (7.4 %) comparing to women (1.1 %). Simi-
lar findings regarding gender specific role of the F2
20210A, as significant risk factor of stroke in men but
not in women, were reported previously [6]. We also
found the evidence that the FV Leiden mutation is ob-
served more frequently in women. A trend toward dif-
ference in the distribution of the F5 1691A was found
between women (5.7 %) and men (1.1 %) from the case
group.
We assume that these data, showing the gender-
specific prevalence of the F2 20210A and F5 1691A
mutations in the case group, suggest that the risk of
stroke development for men and women, carrying the
F2 20210A and F5 1691A mutations, is unequal. There
are extensive epidemiological data showing that pre-
menopausal females have reduced risk for cardiovas-
cular disease than men of the same age, but the meno-
pause initiates a phase of increased risk for cardiovas-
cular disease (CVD) [16– 19].
It was shown that endogenous estrogens increase
the resistance to activated protein C regardless of the
presence of the FVL mutation [6]. At the same time, se-
veral beneficial vascular effects of estrogen have been
suggested, including modification of the composition
of circulating lipoproteins, changes in blood coagula-
tion, inhibition of intravascular accumulation of col-
lagen, antiproliferative effects on vascular smooth
muscle (VSM) cells, and direct vasodilation of blood
vessels [20], and there are few reports on cardiac genes
regulated by estrogen [21].
With age the estrogen concentration in plasma de-
creases, this may explain the enhanced vascular con-
traction in aging females. Postmenopausal hypoestro-
genism has been suggested to increase arterial vascular
tone through a decrease in vasodilator neuropeptides
and an increase in vasoconstrictor peptides in the arteri-
al wall nerve terminals of the autonomous nervous sys-
tem [22].
Progesterone is expressed in both endothelial and
vascular smooth muscle cells, stimulates endothelial
nitric oxide synthase activity, NO production, and NO-
mediated relaxation, the effects of progesterone on
vascular relaxation may be tissue specific [19]. The
FVL mutation has been shown to further increase the
risk of venous thrombosis in oral contraceptive and
hormone replacement therapy users [16, 19].
Nevertheless, some evolutionary benefits for fema-
les carrying the FVL mutation, which are crucial for
successful pregnancy outcome, were described by Go-
pel et al. [23], Majerus et al. [24], and Lindqvist et al.
[25]. Women, who carry the FVL mutation lose less
blood in menstruation, have higher haemoglobin le-
vels, and possibly have a lower incidence of life-
threatening post-partum haemorrhage [25]. The posi-
tive effect on implantation was described [24] and
verified in studies, where an improved implantation
rate in the ICSI and IVF pregnancies was reported if
either the mother or the child carried the FVL mutation
[23]. Those benefits for pre-menopausal women be-
come a disadvantage with age in post-menopause
women with the loss of cardio protective function of
estrogens.
Androgens have been shown to be important for
survival, number of studies have linked androgen defi-
ciency to increased mortality in men [26, 27]. Testoste-
rone is an anabolic hormone with a wide range of bene-
ficial effects on men’s health. Androgen receptors have
been identified in the blood vessels of experimental
animals and of humans [26]. In number of studies it was
[26, 27] noted that testosterone (T) deficiency might
contribute to increased risk of CVD. It was found that
men older than 25 years old presenting with
hypertension had lower total T values [26]. The
androgen deficiency was shown to be associated with
increased levels of total cholesterol, low-density
lipoprotein, increased production of proinflammatory
factors, and increased thickness of the arterial wall and
contributes to endothelial dysfunction [26, 27]. It was
reported that endogenous T levels decline with age.
Thus, we suppose that endothelium dysfunction is
increased in aging men due to testosterone level
decline, so that it can be considered as an independent
thrombosis risk factor. The observed higher frequency
of the F2 20210A variant in men comparing to women
with stroke confirms that the F2 20210A variant might
be an additional genetic risk factor of the stroke deve-
lopment in men.
Furthermore, we have established that, the MTHFR
677T homozygous carrier’s frequency in the case
group in our study (12.6 %) was significantly higher
than in the control group I (5 %). Genotype frequency
302
TATARSKYY P. F. ET AL.
with at least one risk allele of the MTHFR 677T was
higher in women from the case group (54.5 %) than in
women from the control group II (41.5 %).
So, we can assume that carrying the MTHFR 677T,
that results in total homocysteine (tHcy) level increase
[28–30], is a risk factor for the stroke development in
women. As mentioned earlier, endothelium dysfunc-
tion is increased due to sex hormones level decline with
age. Homocysteine may impair endothelium function
by decreasing both the production of vasodilator sub-
stances and their bio-availability to smooth muscle
cells [30]. The damaged blood vessels endothelium in-
creases thrombocyte adhesion and risk of thrombotic
event. Many factors are related to circulating tHcy con-
centrations [29]. Fasting plasma tHcy concentration is
consistently higher in men than in women, and in-
creases with age [22, 29]. In addition, homocysteine
levels are reported to be lower in pre-menopausal wo-
men than in men and post-menopausal women. Be-
cause tHcy concentrations are strongly modulated by
gender and age, interactions among these factors, folate
status and the C677T mutation in the MTHFR gene lo-
cus may explain the inconsistent association of this po-
lymorphism with the risk of occlusive vascular disease.
Changes in sex hormones and tHcy levels seem to have
strongly pronounced influence on endothelium dys-
function development in women. Thus, carrying the
C677T mutation in MTHFR gene may additionally in-
crease the risk of endothelium dysfunction, and as a
result, the risk of stroke development in women with
the 677T allele is higher comparing to men.
For revealing a possible cumulative influence of
the polymorphic variants G20210A, G1691A and
C677T of F2, F5 and MTHFR genes, combined geno-
types distribution analysis was performed (tabl. 2).
After combined genotypes distribution analysis of
the polymorphic variants G20210A, G1691A and
C677T of F2, F5 and MTHFR genes, total frequency of
unfavorable combined genotypes, which included at
least one of the risk alleles of F2 and/or F5 and/or
bearing two copies of the MTHFR 677T, was higher in
men from the case group (20 %) than in men from the
control group II (5.5 %). In women from the case group
(56.8 %) frequency of total unfavorable combined
genotypes which included at least one of the F2, F5,
MTHFR genes risk alleles was higher than in women
from the control group II (41.5 %).
Therefore, our findings suggest some common
trend for interactions of unfavorable combined geno-
types in complex disorder development. The genetic
factors which are minor or insignificant, when present
alone, can exert an additive effect. Certain specific pa-
irs of mutations and clinical risk factors in combination
might yield significant complex risk factors for ische-
mic stroke. Besides the classical clinical risk factors,
rapidly increasing knowledge of unfavorable genetic
303
ISCHEMIC STROKE IN UKRAINIAN POPULATION
Gender
Genotype Case group
Control group
II
F2 F5 MTHFR n % n %
Men
GG GG CC
32 33.7 13 37.3
Women 38 43.2 31 58.5
Men†
GA GG CC
1 1.05 0 0
Women† 0 0 0 0
Men†
GG GA CC
0 0 0 0
Women† 2 2.3 0 0
Men
GG GG CT
44 46.3 20 57.2
Women† 35 39.8 14 26.4
Men†
GA GG CT
5 5.25 0 0
Women† 1 1.1 0 0
Men†
GG GA CT
0 0 0 0
Women† 2 2.3 3 5.7
Men†
GG GG TT
11 11.6 2 5.5
Women† 9 10.2 5 9.4
Men†
GA GG TT
1 1.05 0 0
Women† 0 0 0 0
Men†
GG GA TT
1 1.05 0 0
Women† 1 1.1 0 0
Men
GA or/and GA
or/and TT
19 20* 2 5.5
Women
GA or/and GA
or/and CT + TT
50 56.8* 22 41.5
n – group size; F2: A – 20210A; G – 20210G, F5: A – 1691A; G –
1691G, MTHFR: T – 677T, C – 677C; † – genotypes, considered as
unfavorable; *Significant at P < 0.05.
Table 2
Combined genotypes frequencies of the F5 G1691A, F2 G20210A
and MTHFR C677T genes variants
mutations will permit recognition of a larger popula-
tion at high risk of ischemic stroke, and this may lead to
more effective prevention.
We should point out, that larger case and control
groups for verification of the observed interactions in
our study in higher significant level are required.
Our data show that the polymorphic variant
20210A of F2 gene can be a higher risk factor for the
stroke development in men. On the other hand, for wo-
men a higher risk factor for the stroke development se-
ems to be the polymorphic variant 1691A of F5 gene.
We also assume that the polymorphic variant 677T of
MTHFR gene can be a genetic predisposition factor for
the stroke development in women. Cumulative risk fac-
tor for the stroke development is revealed in a combi-
nation of unfavorable polymorphic variants 20210A,
1691A and 677T of the F2, F5 and MTHFR genes.
The study was supported by the NAS of Ukraine.
Ï. Ô. Òà òà ðñüêèé, À. Ì. Êó ÷å ðåí êî, Ñ. À. Êðàâ ÷åí êî,
Ä. Â. Øóëü æåí êî, Ñ. Ì. Êóçíºöîâà, Ë. À. ˳âøèöü
²øåì³÷íèé ³íñóëüò ñå ðåä íà ñå ëåí íÿ Óêðà¿ íè: ìîæ ëè âå
çà ëó ÷åí íÿ ïîë³ìîð ôíèõ âàð³àíò³â ãåí³â F2 G20210A,
F5 G1691A òà MTHFR C677T
Ðå çþ ìå
Ìåòà. Ïðî âåñ òè àíàë³ç çâ’ÿç êó ïîë³ìîðô ííèõ âàð³àíò³â ãåí³â
F2, F5, MTHFR òà ïà òî ãå íå çó ³íñóëü òó ñå ðåä íà ñå ëåí íÿ Óêðà¿-
íè. Ìå òî äè. Àëåëüí³ âàð³àíòè âèç íà ÷à ëè ñå ðåä 183 íå ñïîð³äíå -
íèõ ïàö³ºíò³â ç ³íñóëü òîì, 100 ³íäèâ³äóóì³â ³ç çà ãàëü íî¿ ïî ïó ëÿ-
ö³¿ òà 88 – çäî ðî âèõ ³íäèâ³äóóì³â ñòàð øèõ 65 ðîê³â, ìå òî äîì
ÏËÐ òà ÏÄÐÔ-àíàë³çó. Ðå çóëü òà òè. «Íåñ ïðè ÿò ëèâ³» ïîë³ìîð-
ôí³ âàð³àíòè F2 20210A, F5 1691A òà MTHFR 677T ñïîñ òåð³ãà -
ëè ñÿ ÷àñò³øå ñå ðåä ïàö³ºíò³â ç ³øåì³÷íèì ³íñóëü òîì ïîð³âíÿ íî
ç êîí òðîëü íè ìè ãðó ïà ìè. Âèñ íîâ êè. Ïîë³ìîðôí³ âàð³àíòè F5
1691A òà MTHFR 677Ò àñîö³éî âàí³ ç âè ïàä êà ìè ³øåì³÷íî ãî ³í-
ñóëü òó ó æ³íîê, âàð³àíò F2 20210A – ç âè ïàä êà ìè ³øåì³÷íî ãî
³íñóëü òó ó ÷î ëîâ³ê³â. Êîìá³íàö³ÿ âè ÿâ ëå íèõ ìó òàí òíèõ âàð³àí-
ò³â 1691À, 20210À ³ 677T ãåí³â F5, F2 ³ MTHFR ïðî ÿâ ëÿºòüñÿ ÿê
àäè òèâ íèé ôàê òîð ðè çè êó âè íèê íåí íÿ èí ñóëü òó.
Êëþ ÷îâ³ ñëî âà: ãåí ôàê òî ðà V Ëåé äåí (F5 G1691A), ãåí ïðî -
òðîìá³íó (F2 G20210A), ãåí ìå òè ëåí òåò ðàã³äðî ôî ëàò ðå äóê -
òàçè (MTHFR Ñ677Ò), ³øåì³÷íèé ³íñóëüò, êîìá³íî âà íèé
ãå íî òèï.
Ï. Ô. Òà òàð ñêèé, À. Ì. Êó ÷å ðåí êî, Ñ. À. Êðàâ ÷åí êî,
Ä. Â. Øóëü æåí êî, Ñ. Ì. Êó çíeöî âà, Ë. À. Ëèâ øèö
Èøå ìè ÷åñ êèé èí ñóëüò ñðå äè íà ñå ëå íèÿ Óêðà è íû: âîç ìîæ íîå
âîâ ëå ÷å íèå ïî ëè ìîð ôíûõ âà ðè àí òîâ ãå íîâ F2 G20210A, F5
G1691A è MTHFR C677T
Ðå çþ ìå
Öåëü. Ïðî âåñ òè àíà ëèç àñ ñî öè à öèè ïî ëè ìîð ôíûõ âà ðè àí òîâ
ãå íîâ F2, F5, MTHFR è ïà òî ãå íå çà èí ñóëü òà ñðå äè íà ñå ëå íèÿ
Óêðà è íû. Ìå òî äû. Àëëåëüíûå âà ðè àí òû àíà ëè çè ðî âà ëè ñðåäè
183 íå ðî äñòâåí íûõ ïà öè åí òîâ ñ èí ñóëü òîì, 100 èí äè âè äó ó ìîâ
èç îá ùåé ïî ïó ëÿ öèè Óêðà è íû è 88 çäî ðî âûõ èí äè âè äó ó ìîâ
ñòàð øå 65 ëåò ìå òî äà ìè ÏÖÐ è ÏÄÐÔ-àíà ëè çà. Ðå çóëü òà òû.
«Íåá ëà ãîï ðè ÿò íûå» ïî ëè ìîð ôíûå âà ðè àí òû F2 20210A, F5
1691A è MTHFR 677T âû ÿâ ëÿ ëèñü ÷àùå ó ïà öè åí òîâ ñ èøå ìè -
÷åñ êèì èí ñóëü òîì ïî ñðàâ íå íèþ ñ êîí òðîëü íû ìè ãðóï ïà ìè.
Âû âî äû. Ïî ëè ìîð ôíûå âà ðè àí òû F5 1691A è MTHFR 677Ò àñ -
ñî öè è ðî âà íû ñî ñëó ÷à ÿ ìè èøå ìè ÷åñ êî ãî èí ñóëü òà ó æåí ùèí,
âà ðè àíò F2 20210A – ñî ñëó ÷à ÿ ìè èøå ìè ÷åñ êî ãî èí ñóëü òà ó
ìóæ ÷èí. Êîì áè íà öèè âû ÿâ ëåí íûõ ìó òàí òíûõ âà ðè àí òîâ
1691À, 20210À è 677T ãå íîâ F5, F2 è MTHFR ïðî ÿâ ëÿ þò ñÿ êàê
àä äè òèâ íûé ôàê òîð ðèñ êà âîç íèê íî âå íèÿ èí ñóëü òà.
Êëþ ÷å âûå ñëî âà: ãåí ôàê òî ðà V Ëåé äåí (F5 G1691A), ãåí
ïðî òðîì áèía (F2 G20210A), ãåí ìå òè ëåí òåò ðà ãèä ðî ôî ëàò-
ðå äóê òà çû (MTHFR Ñ677Ò), èøå ìè ÷åñ êèé èí ñóëüò, êîì áè íè ðî -
âàí íûé ãå íî òèï.
REFERENCES
1. Madonna P., de Stefano V., Coppola A., Cirillo F., Cerbone
A. M., Orefice G., Di Minno G. Hyperhomocysteinemia and
other inherited prothrombotic conditions in young adults with
a history of ischemic stroke // Stroke.–2002.–33, N 1.–P. 51–
56.
2. Meyer M. R., Haas E., Barton M. Gender differences of car-
diovascular disease: new perspectives for estrogen receptor
signaling // Hypertension.–2006.–47, N 6.–P. 1019–1026.
3. Markus H. Genes for stroke // J. Neurol. Neurosurg. Psych³at-
ry.–2004.–75, N 9.–P. 1229–1231.
4. Margaglione M., D’Andrea G., Giuliani N., Brancaccio V.,
De Lucia D., Grandone E., De Stefano V., Tonali P. A., Di
Minno G. Inherited prothrombotic conditions and premature
ischemic stroke: sex difference in the association with factor
V Leiden // Artherioscler. Thromb. Vasc. Biol.–1999.–19,
N 7.–P. 1751–1756.
5. De Stefano V., Chiusolo P., Paciaroni K., Casorelli I., Rossi
E., Molinari M., Servidei S., Tonali P. A., Leone G. Prothrom-
bin G20210A mutant genotype is a risk factor for cerebrovas-
cular ischemic disease in young patients // Blood.–1998.–91,
N 10.–P. 3562–3565.
6. Lalouschek W., Schillinger M., Hsieh K., Endler G., Tents-
chert S., Lang W., Cheng S., Mannhalter C. Matched case-
control study on factor V Leiden and the prothrombin
G20210A mutation in patients with ischemic stroke/transient
ischemic attack up to the age of 60 years // Stroke.–2005.–36,
N 7.–P. 1405–1409.
7. Bertina R. M., Koeleman B. P., Koster T., Rosendaal F. R.,
Dirven R. J., de Ronde H., van der Velden P. A., Reitsma P. H.
Mutation in blood coagulation factor V associated with resis-
tance to activated protein C // Nature.–1994.–369, N 6475.–
P. 64–67.
8. Ridker P. M., Hennekens C. H., Lindpaintner K., Stampfer M.
J., Eisenberg P. R., Miletich J. P. Mutation in the gene coding
for coagulation factor V and the risk of myocardial infarction,
stroke, and venous thrombosis in apparently healthy men // N.
Engl. J. Med.–1995.–332, N 14.–P. 912–917.
9. Poort S. R., Rosendaal F. R., Reitsma P. H., Bertina R. A
common genetic variation in the 39-untranslated region of the
304
TATARSKYY P. F. ET AL.
prothrombin gene is associated with elevated plasma pro-
thrombin levels and increase in venous thrombosis // Blood.–
1996.–88, N 10.–P. 3698–3703.
10. Maniatis T., Fritsch E. F., Sambrook J. Molecular cloning: a
laboratory manual.–New York: Cold Spring Harbor Lab.
publ., 1982.–545 p.
11. Koksal V., Baris I., Etlik O. Primer-engineered multiplex
PCR–RFLP for detection of MTHFR C677T, prothrombin
G20210A and factor V Leiden mutations // Exp. Mol. Pa-
thol.–2007.–83, N 1.–P. 1–3.
12. Michael S., Qamar R., Akhtar F., Khan W. A., Ahmed A.
C677T polymorphism in the methylenetetrahydrofolate re-
ductase gene is associated with primary closed angle glauco-
ma // Mol. Vis.–2008.–14.–P. 661–665.
13. Moore J. H. Computational analysis of gene-gene interacti-
ons using multifactor dimensionality reduction // Expert Rev.
Mol. Diagnost.–2004.–4, N 6.–P. 795–803.
14. Raymond M., Rousset F. GENEPOP (version 1.2): popula-
tion genetics software for exact tests and ecumenicism // J.
Hered.–1995.–86, N 3.–P. 248–249.
15. Excoffier L., Laval G., Schneider S. Arlequin ver. 3.0: An in-
tegrated software package for population genetics data analy-
sis // Evol. Bioinform. Online.–2005.–1.–P. 47–50.
16. Henkens C., Bom V., Seinen A., van der Meer J. Sensitivity to
activated protein C: influence of oral contraceptives and sex
// Thromb. Haemost.–1995.–73, N 3.–P. 402–404.
17. Jensen-Urstad K., Johansson J. Gender difference in age-re-
lated changes in vascular function // J. Int. Med.–2001.– 250,
N 1.–P. 29–36.
18. Stampfer M. J., Colditz G. A., Willett W. C. Menopause and
heart disease. A review // Ann. N. Y. Acad. Sci.–1990.–
592.–P. 193–203.
19. Koledova V. V., Khalil R. A. Sex hormone replacement thera-
py and modulation of vascular functionin cardiovascular
disease // Expert Rev. Cardiovasc. Ther.–2007.–5, N 4.–
P. 777–789.
20. Orshal J. M., Khalil R. A. Gender, sex hormones, and vascu-
lar tone // Am. J. Physiol. Regul. Integr. Comp. Physiol.–
2004.–286, N 2.–P. R233–R249.
21. Otsuki M., Gao H., Dahlman-Wright K., Ohlsson C., Eguchi
N., Urade Y., Gustafsson J. A. Specific regulation of lipoca-
lin-type prostaglandin D synthase in mouse heart by estrogen
receptor beta // Mol. Endocrinol.–2003.–17, N 9.–P. 1844–
1855.
22. Di Carlo C., Di Spiezio Sardo A., Bifulco G., Tommaselli G.
A., Guerra G., Rippa E., Mandato V. D., Nappi C. Postmeno-
pausal hypoestrogenism increases vasoconstrictor neuropep-
tides and decreases vasodilator neuropeptides content in arte-
rial-wall autonomic terminations // Fertil. Steril.–2007.–88,
N 1.–P. 95–99.
23. Gopel W., Ludwig M., Junge A. K., Kohlmann T., Diedrich
K., Moller J. Selection pressure for the factor-V-Leiden mu-
tation and embryo implantation // Lancet.–2001.–358,
N 9289.–P. 1238–1239.
24. Majerus P. W. Human genetics. Bad blood by mutation //
Nature.–1994.–369, N 6475.–P. 14–15.
25. Lindqvist P. G., Zoller B., Dahlback B. Improved hemoglobin
status and reduced menstrual blood loss among female carri-
ers of factor V Leiden an evolutionary advantage? // Thromb.
Haemost.–2001.–86, N 4.–P. 1122–1123.
26. Liu P. Y., Death A. K., Handelsman D. J. Androgens and car-
diovascular disease // Endocrinol. Rev.–2003.–24, N 3.–
P. 313–340.
27. Traish A. M., Saad F., Feeley R. J., Guay A. The dark side of
testosterone deficiency: III. Cardiovascular disease // J.
Androl.–2009.–30, N 5.–P. 477–494.
28. Russo G. T., Friso S., Jacques P. F., Rogers G., Cucinotta D.,
Wilson P. W., Ordovas J. M., Rosenberg I. H., Selhub J. Fra-
mingham Offspring Study Cohort. Age and gender affect the
relation between methylenetetrahydrofolate reductase C677T
genotype and fasting plasma homocysteine concentrations in
the Framingham Offspring Study Cohort // J. Nutr.–2003.–
133, N 11.–P. 3416–3421.
29. Dierkes J., Jeckel A., Ambrosch A., Westphal S., Luley C.,
Boeing H. Factors explaining the difference of total homocys-
teine between men and women in the European Investigation
Into Cancer and Nutrition Potsdam Study // Metabolism.–
2001.–50, N 6.–P. 640–645.
30. Woo K. S., Chook P., Lolin Y. I., Cheung A. S., Chan L. T.,
Sun Y. Y., Sanderson J. E., Metreweli C., Celermajer D. S.
Hyperhomocyst(e)inemia is a risk factor for arterial endothe-
lial dysfunction in humans // Circulation.–1997.–96, N 8.–
P. 2542–2544.
ÓÄÊ 575.11 + 577.21
Received 20.03.10
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