**3. Experiment design**

In our experiments we studied the indicators of lipid and lipoprotein metabolism in the blood plasma and the liver under the experimental metabolic syndrome (MS) in Syrian hamsters of different sex and age.

In the experiments purebred male rats with 180-220 g of the body weight were used. The animals were kept in vivarium on a balanced diet. During 21 days the animals were given low alcoholic beverages from grapes of red and white grades *per os* daily. These beverages were introduced in the maximum effective doses of 9 mg of polyphenols/100 g the body weight. Taking into account the fact that the polyphenol content in the beverages investigated was quite low, the effective dose was introduced 3 times a day by 2 ml of liquids per 100 g of the animal's body weight. Control animals were introduced the corresponding volume of the saline solution. Ethanol was given in the corresponding dose.

450 Lipoproteins – Role in Health and Diseases

the LDL uptake by macrophages, therefore, it inhibits atherogenesis. This allows suggesting that the therapeutic effect of the medicine may not be connected with its ability to decrease the

The medicine is absorbed slowly when taken internally, it is readily soluble in the adipose tissue releasing gradually into the bloodstream, and so its action is kept for a long time (up

When using probucol in MultiVitamins and Probucol (MVP) research the renewal of the endothelium function in patients with IHD, decrease of restenosis cases after coronary angioplasty (when taking it at least 4 weeks before the procedure and further treatment during 6 months) was observed. Other antioxidants (α-tocopherol in high doses (700 mg per

Combined application of the endogenous antiradical antioxidants is of particular interest. In HPS (Heart Protection Study) along with the study of the simvastatin effectiveness the prophylactic action of antioxidants was investigated. The use of the vitamin complex (600 mg of vitamin E, 250 mg of vitamin C and 20 mg of β-carotene per day) lasted in average 5.5 years and did not reveal any differences in placebo groups and groups taking vitamins. Moreover, if the tendency exists, it reflects increasing of vascular events in the antioxidant intent-to-treat group. The action of antioxidants was compared with the effect of the combined use of simvastatin and nicotinic acid (niacin). Moreover, one of the groups received simvastatine+niacin and antioxidants. Angiographic and clinical data of this study

Unfortunately, a great part of the compounds synthesized, which are used for pharmacocorrection of these states, are xenobiotics, so they can activate the free-radical formation process. Synthetic antioxidants, in particular probucol, can not be recommended

The lack of antioxidant medicines popularity and the absence of traditions of their common use in practical medicine are caused a number of reasons: unsatisfactory previous study of this issue, complexity of adequate estimation of oxidation state parameters in the organism and the absence of the effective medicines with the antioxidant activity that are able to

Therefore, the main indications for using antioxidants are excessively activated free-radical oxidation processes accompanying different pathologies. The choice of specific medicines, correct indications and contraindications for their use has not been developed yet and

In our experiments we studied the indicators of lipid and lipoprotein metabolism in the blood plasma and the liver under the experimental metabolic syndrome (MS) in Syrian

LDL level. There is no clinical evidence of this hypothesis at the moment.

to 6 months after discontinuation of the treatment).

day), β-carotene and vitamin C) turned out to be ineffective.

were also disappointing with respect to the use of antioxidants.

for patient use because they decrease the HDL-C level.

quickly reduce the consequences of the oxidative stress.

require further research.

**3. Experiment design** 

hamsters of different sex and age.

Stress was caused by immobilization on the abdomen for 3 hours [19]. Animals were decapitated 3 hours after the immobilization. The blood was collected to get the serum. The liver was perfused by the cold extraction medium (0.25 M sucrose in 0.025 M tris-HCl, pH 7.5), homogenized in the Potter homogenizer with 2 ml of the extraction medium per 1 g of the liver. All manipulations with animals were held under chloralose-urethane anaesthesia.

To distribute the plasma lipoproteins the samples were centrifuged at 65,000 rpm (342,000 *g*) for 4 h at 4°C in the Optima XL-100K ultracentrifuge (Beckman Coulter) set at slow acceleration and deceleration [20]. Samples were fractionated within 1 h of centrifugation.

Lipids were extracted with chloroform and methanol (1:2 v/v) twice, as described by Bligh et al [21], and the supernatant was collected for determination of TG and FFA. TG and FFA were determined by enzymatic colorimetric methods with commercial kits (Zhongsheng, Beijing, China). The total cholesterol content was detected with the help of standard enzymatic cholesteroloxidase kits of "Boehringer Mannheim GmbH diagnostica" firm (Germany). The total lipid concentration was determined with the help of a standard kit "Eagle Diagnostics" (USA) – the reaction with vanillin reagent.

Determination of the lipid peroxide product quantity was performed in heptaneisopropanol extracts [22]. The optical density was measured at the wavelength of 220 nm (for compounds with isolated double bonds), 232 nm (for diene conjugates) and 278 nm – for ketodienes and conjugate trienes.

The TBA content was determined on the spectrophotometer with the help of the reaction with thiobarbituric acid [23].

A modified version of the high performance liquid chromatography (HPLC) procedure developed by Stacewicz-Sapuntzakis et al. [24] was used to measure vitamins E in the plasma. The HPLC system included a 150 × 3.9 mm Nova-pak C18 (4 microns) column with a guard-pak pre-column (both from Waters, Milford, MA), Waters Millipore TCM column heater, Waters 490 multi-wavelength detector, Hitachi 655–61 processor, Hitachi 655A-11 liquid chromatography, and BioRad autosampler AS-100.

The serum ascorbic acid concentrations were measured as described by using HPLC [25] with salicylsalicylic acid as a deproteinizing agent, metaphosphoric acid as a stabilizer.

The serum PON1 activity was measured by the rate of generation of p-nitrophenol determined at 405 nm according to MacKness B et al. [26].

The plasma cholesterol ester transfer protein (CETP) activity was examined using the modifications of Khosla et al. [44]. The CETP activity in duplicate10-μL aliquots of the plasma was determined after incubations with 3H-cholesterol ester (CE)-labeled HDL3 and LDL. Radioactivity transferred from 3H-HDL3 to LDL (measured in the supernatant after precipitation with heparin/MnCl2+) was used to calculate the CETP activity (expressed as the percentage of radioactivity transferred from 3H-HDL3 to LDL per 16 h of incubation).

To measure endothelium-bound LPL, the perfusion solution was changed to buffer containing 1% fatty acid–free BSA and heparin (5 units/ml). The coronary effluent was collected in timed fractions over 10 min and assayed for the LPL activity by measuring the hydrolysis of a sonicated [3H]triolein substrate emulsion [27].

The plasma LCAT activity was measured by determination of the amount of radioactivity in each spot calculating the free cholesterol/ total cholesterol ratio in each plasma sample before and after the LCAT reaction and thus estimating the esterification rate [28]. The fractional esterification rate *(%* . h') expressed as the percentage of the free cholesterol esterified in the plasma sample per hour.

The HL activity was evaluated using the glycerol-stabilized emulsion of triolein and egg phosphatidylcholine containing glycerol-tri[9,10(n)-3H] oleate by determination of the radioactivity amount during incubation [29].

**Statistical analysis**. All data were analyzed for statistical significance with SPSS 13.0 software. The data were presented as means ± standard deviation. Statistical analysis used one-way ANOVA. P<0.05 was considered to be statistically significant.
