The importance of MTHFR, MTR, MTRR

and CSE expression levels in Caucasian

women with preeclampsia

Agnieszka Seremak-Mrozikiewicz, Anna Bogacz,

Joanna Bartkowiak-Wieczorek, Hubert Wolski,
Boguslaw Czerny, Malgorzata Gorska-Paukszta,
Krzysztof
Drews and Reproductive Biology 188 (2015) 113
European Journal of Obstetrics
& Gynecology

Introduction

Introduction
Methionine and homocysteine metabolism

(MHM):
Methylenetetrahydrofolate reductase

(MTHFR)
Methionine synthase (MTR)
Methionine synthase reductase (MTRR) and
CSE
Normal pregnant women had low plasma

concentrations of homocysteine during

gestation whereas hyperhomocysteinemia
occurred in women with PE

Introduction

Materials and methods

Patients
Caucasian of Polish origin recruited in Division of

Perinatology and Womens Diseases, Poznan

University of Medical Sciences
90 pregnant women with PE (29 women with mild
PE and 61 women with severe PE)
control group included 74 healthy pregnant
women
Exclusion criteria : diabetes mellitus, obesity,
cardiovascular diseases, chronic hypertension,
renal and endocrinal diseases, thromboembolism,
age lower than 18 years, multifetal pregnancy.

Materials and methods

Data collection
In each woman the blood pressure was

measured sitting or lying with the

sphygmomanometer (twice with 6 h interval).
The urine sample was analyzed and the
proteinuria was recognized by presence of
protein 30 mg/dl in sample (or in test strip as
result 1+).
Immediately after birth the fragment from the
central part of placenta was collected
sampled in maternal side in order to obtain
villous and decidua

Materials and methods

Laboratory analysis
Total cellular RNA was isolated from the

placenta tissue using TriPure Isolation Reagent

Concentrations and the purity of RNA were
determined by measuring the absorbance at
260 and 280 nm in a spectrophotometer
Complementary DNA was synthesized from 2
mg of total RNA in a total volume of 20 ml using
the Transcriptor First Strand cDNA Synthesis Kit
Level of mRNA expression was analyzed by
using RT-PCR

Table 1 Sequences of primers used for the RT-PCR analysis and

PCR conditions

Materials and methods

Statistical Analysis
mRNA content for studied genes was

expressed as mean SEM.

The experimental data were analyzed using
the SPSS 17.0 for Windows software.
Mean values were compared by one-way
ANOVA test.
The value of p < 0.05 was considered as
statistically significant.

Results

Results

Fig. 1. The placental expression level of MTHFR, MTR, MTRR, and

CTH in pregnant women with PE. Control group was defined as
100%. Data were presented as mean SEM.
*** p < 0.0001 as compared to the control group (one-way
ANOVA test).

Results

Fig. 2. The placental expression level of MTHFR, MTR, MTRR, and CTH in
pregnant women with mild and severe PE. Control group was defined as
100%. Data were presented as mean SEM. * p < 0.05, ** p < 0.001, *** p
< 0.0001 as compared to the control group (one-way ANOVA test).

DISCUSSION

DISCUSSION
Sepulveda et al
Increase of mrna level of MTR in placental

tissue obtained from patients with PE

Associated with decrease 2methoxyestradiol (2-ME) synthesized by
cathechol-o-methyl-transferase (COMT)
that induces the differentiation of the
endovascular cytotrophoblast cells in the
presence of hypoxia
Low levels of circulating 2-ME at the first
trimester of pregnancy in patients who
later developed preeclampsia

DISCUSSION
Kanasaki et al.
Similar effect on low levels of 2-ME in women
with PE was observed during the third trimester
of pregnancy
Not only a decrease of COMT expression but
alterations in the methionine and
homocysteine metabolism could be involved in
the pathogenesis of PE
SAM plays an important role because acts as
the principal methyl donor within cells
Low levels of SAM may be caused by a
decreased activity of MTHFR and/or MTR

DISCUSSION
Decrease of MTRR mRNA level may be

caused by many factors such as diet rich in

folic acid and vitamin B12, administration
of the drugs (methyldopa).
A slight increase of the placental CSE
expression level between preeclamptic
women and control group indicates that
homocysteine may be eliminated through
the alternate transsulfuration pathway
which is supported by the increased mRNA
levels of CSE and/or CBS.

Conclusions

CRITICAL APPRAISAL

CRITICAL APPRAISAL
Questions

Answers

Explanation

1. Study design,
survey or registration?

Survey

See page 1:
The control group consisted of 74
healthy pregnant women and 90
patients with diagnosed pre-eclampsia.
Total RNA was isolated from placenta
and the mRNA level of examined
genes was to determine using realtime PCR.

2. Inductive or
deductive reasoning?

Deductive
(theory
prediction
observation
conclusions)

See page 1-2:

Theory: Writers explained the theory of
vasodilator factors that play an
important role in genetic background
of PE.
Prediction: Changes in the expression
levels of enzymes involved in the MHM
cycle may be related to development
of PE
Conclusions: Elevated RNA expression
of MTR in placenta of preeclamptic
patients is probably results of a
potential compensation mechanism of

CRITICAL APPRAISAL
Questions
3 and 4 Type and
scales of each
variables in this
study

Answers

Explanation
Dependent variables:
Preeclampsia (nominal)
Independent Variables:
Age (ratio)
Gestational age at delivery (ratio)
Blood pressure (ratio)
Height (ratio)
Body wght before and after
pregnancy (ratio)
Infant birth weight (ratio)
Number of pregancy (ratio)
Delivery mode (nominal)
Hemoglobin (ratio)
Eritrocytes (ratio)
Leukocytes (ratio)
Platelets (ratio)
Hematocrit (ratio)
Protein in urine (ratio)
Levels of MTHFR, MTR, MTRR, CTH

CRITICAL APPRAISAL
Questions
5. Type of data,
primary, secondary
or tertiary
resources?

Answers
Primary

Explanation
Blood pressure, protein urine, and
placental fragments was obtained
during research

6. Group or ungroup Ungroup

data?

Data was taken primarily and

analyzed by researchers

7. Ad hoc or
routine data?

Ad hoc

Data was collected for some

period of time during study

8. Measures of
Central Tendency,
Position and
Dispersion

Central
Tendency :
Mean
Dispersian:
SEM

Some variables expressed as

Mean SEM

CRITICAL APPRAISAL
Questions

Answers

Explanation

9. Tables that was Frequency

used to present
tables
results of this
study

See page 114 and 115

10. Graph used in Histogram

this paper

See page 116

CRITICAL APPRAISAL
Questions
11. Quality of
research data

Answers
High
quality

Explanation
1. Used primary data
2. Good operational definition
3. Using valid measurement
technique

CRITICAL APPRAISAL
Questions

Answers

Explanation

12. Bias

Low risk of
bias

Almost all known confounding

factors was controlled

13. Sample size

calculation for
this study

Not
mentioned

14. Sampling
technique

Total
sampling

All studies that met inclusion and

exclusion criteria were included
in this study

CRITICAL APPRAISAL
Questions
15 and 16.
Statistical
analysis

Answers
Bivariate
analysis:
one-way
ANOVA test
to compare
mean
values

Explanation
The experimental data were
analyzed using the SPSS 17.0 for
Windows software. Mean values
were compared by one-way
ANOVA test. The value of p < 0.05
was considered as statistically
significant.

17. Error to
None
conclude
statistical analysis
results
18. How was the
presentation of
results in this
paper?

Good

Writers clearly explained

background, methods and
results in this study. They
compare results with another

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