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Page 4 of 9 Nemtsova et al. Vessel Plus 2018;2:27 I http://dx.doi.org/10.20517/2574-1209.2018.48

Determination of total cholesterol (TC), triglycerides (TG) and cholesterol of high density lipoproteins
(HDL cholesterol) were performed in serum enzymatically by photocolorimetric method with sets
produced by Human (Germany). The content of cholesterol in the low density lipoprotein (LDL cholesterol)
was calculated by the formula of Friedewald W. T. with consideration of measurement in mmol/L: LDL
cholesterol = cholesterol - (HDL cholesterol + TG/2.22).

Determination of the concentration of fasting glucose was performed by the glucose oxidase method using
analyzer Humolizer (made in Germany). The level of glycated hemoglobin (HbA1c) was measured by
enzyme immunoassay (ELISA) using a set of reagents Hummer (USA). To determine the insulin resistance
(IR) index HOMA-IR was used, which was calculated with the formula: [(Glucose fasting) × (fasting
insulin)] mmoL/mL/22.5.

Besides the indicators of carbohydrate and lipid metabolism all patients underwent measurement of the
concentration of insulin in blood serum by the method of ELISA using a kits DRG Instrument Gmbh
(Germany) on a semi-automatic ELISA analyzer “ImmunoChem-2100”, HighTechnology, Inc. (USA).

To study the antioxidative system, the activity of glutathione peroxidase (GPO) and the level of sulfhydryl
groups (SH-groups) were assessed. GPO plays an important role in protecting biological cell membranes
against oxidative damage by increasing the concentration of reduced glutathione (oxidised glutathione
ratio - GSSG-R) in the process of aerobic glycolysis. SH-groups are the organic compounds that contain
a sulphydryl group. Among all the antioxidants that are available in the body, they constitute the major
portion of the total body antioxidants and they play a significant role in defense against reactive oxygen
species (ROS). The level of malonic dialdehyde (MDA) was used as a marker of the lipid peroxidation
and oxidative system activity. The activity of GPO (KF 1.11.1.9) in Ethylenediaminetetraacetic acid
(EDTA)-hemolysate was determined by the decrease in the content of reduced glutathione during
a 5-min incubation of a test sample of hemolysate in the presence of oxidizing substrate - cumene
[12]
hydroperoxide by the photometric method . The SH-groups and MDA were determined in serum using
[12]
a photometric method . The following reagents were used: thiobarbituric acid (Organika, Germany),
dithiobisnitrobenzoic acid (Merck, Germany), restored glutathione (Sigma-Aldrich, Germany), cumene
hydroperoxide (Merck, Germany). The determination of 8-hydroxy-2-deoxyguanosine (8-OH-dG) in blood
serum, as one of the biomarkers of oxidative damage, was carried out by ELISA with kits “Bio-Vendor”
(Czech Republic).

Ultrasound examination of the common CIMCT was performed according to the standard procedure on
the device “LOGIQ5”.

The results obtained are presented as the average value of parameters (M) and standard error (m).
Processing of statistical data was performed using the software package “Statistics for Windows 8.0”. The
student criterion and Pearson’s chi-squared test were used to estimate the differences between groups with
normal distribution. The differences were considered statistically significant at P < 0.05.

The study was performed in compliance with the basic provisions of the Helsinki declaration of the world
medical association on ethical principles of scientific and medical research involving humans (1964-2000)
and the order of the ministry of health of Ukraine dated 23.09.2009 № 690. The article is a fragment of
the research work of the department of clinical pharmacology and internal medicine of Kharkiv National
Medical University “Optimization of diagnosis and treatment of comorbid pathology (HT and diabetes
mellitus type 2) on the basis of evaluation of cardio-hemodynamics, metabolism and pharmacogenetic