ABOUT DEUTERIUM DEPLETED WATER INFLUENCE ON INDICATORS OF PROOXIDE-ANTIOXIDE SYSTEM AT LABORATORY ANIMALS

Barishev M.G., Basov A.A., Bolotin S.N., Dzhimak S.S., Dolgov M.A.,

Fedosov S.R., Shashkov D.I.

By nuclear magnetic resonance spectroscopy established decrease in the maintenance of deuterium in blood plasma, erythrocytes and homogenate of hearts at the laboratory rats infected Staphylococcus aureus, using deuterium depleted water within a month. At rats also infected Staphylococcus aureus, using the usual water, any changes of isotope structure it is not revealed. By method EPR spectroscopy it is defined that the quantity of the paramagnetic centers heart and a liver of the rats took deuterium depleted water after lyophilization, has decreased in 4 times in comparison with control group that is testifies about reduction of quantity of free radicals and favorable influence of deuterium depleted water on animals organism.

Deuterium depleted water, NMR spectroscopy, EPR spectroscopy, free radicals, oxidative stress

This work is executed with financial support of the Russian fund of basic researches, the project ¹11-04-96523-r_south_c.

In modern preventive medicine the great attention is given to the correction of metabolic infringements in organism on nonclinic stage for the purpose of the prevention of development of proof complications. One of nonspecific protective systems of the organism is antioxide system (ÀÎS) [1].

It is known that antioxidants regulated normal activity of a human body, in particular processes of lipids oxidation, fibers and nucleinic acids in which result in cages highly active substances of oxygen and free radicals are formed[2,3]. In physiological conditions free radicals are necessary for normal breath, metabolism and destruction of alien bacteria.

At the same time at superfluous strengthening of uncontrollable reactions free radicals oxidations the typical pathological process characterized by strengthened formation of primary radicals (superoxide anion-radical, nitrogen oxide), jet molecules (singlet oxygen, a hydrogen peroxide, hydroperoxides), secondary radicals hydroxyl radical, alkils, alkoxiles, peroxiles) against oppression fermental (superoxidazmutaza can develop, the catalase, glutathione reductase) and non fermental (vitamins, thiols, polyphenols) links of antioxidå systems (ÀÎS) that is accompanied by disbalance of functioning protective organism systems and decrease in its nonspecific resistance to pathological processes [4].

The food is one of the environment factor, essentially influencing to the health, working capacity and life expectancy of the person. Now, besides an optimum parity of food components and mineral substances in a diet, the great attention is given to the influence nutrients to the indicators endogen antioxidå system and level free radicals oxidations in an organism that can promote increase in duration of active ability to live and preventive maintenance of complications of some diseases, in pantogenesis which the essential role is played by the phenomena of oxidizing stress (diabetes, an atherosclerosis, a bronchial asthma, tumoral and inflammatory diseases of various localization, etc.).

The greatest interest is represente the revealing possibility antioxide means, specifically operating on concrete links of processes free radicals of oxidation, and, except that, products interfering superfluous formation peroxide oxidation and the last eliminating toxic effects [5].

Important selection for the further tests of such substances which at introduction in clinical practice, along with high efficiency antioxide actions, would show a minimum by-effects represented.

The water with the lowered maintenance of a deuterium is one of the possible factor that raising working of the OAS.

It is known that the person in physiological conditions consumes 2-3 liters of liquid in a day, therefore changes of structure of a food by the expense of water with the lower maintenance of a deuterium can influence indicators ÀÎS. Biological effects of water with the lower maintenance of a deuterium are described in this article [6].

The purpose of this research is studying influence of water with the lower maintenance of a deuterium (concerning natural level) on indicators prooxide-antioxide system and the quantitative maintenance of a deuterium in fabrics of laboratory rats in one-monthly experiment [7].

Reception gomogenate fabrics carried out by the mechanical crushing of bodies (heart, liver, kidneys) before reception of homogeneous weight with the subsequent in centrifuge at 3000 rpm within 20 minutes. The supernatant received took away by means of a micropipette in volume of 2 ml.

For definition of the quantitative maintenance of free radicals by method EPR spectroscopy in heart, liver and their kidneys subjected liophilic to drying (the device: a dryer liophilic LS-1000).

The definition of the concentration of a deuterium in biological liquids and gomogenat fabrics have been spent on a pulse nuclear magnetic resonance spectrometer JEOL JNM-ECA 400MHz. Shooting of spectra was spent on corresponding resonant frequency of kernels of a deuterium – 61.4 MHz. Shooting parameters: 6.7 with (acquisition time), 20 with (relaxation delay), 5.6 mks (x-pulse), 0.15 Hz (resolution). Shooting temperature – 25 ◦C, thus accuracy of stabilization 0.2 ◦C. Measurements were spent with the using of an ampoule of 5 mm in which has been strictly fixed soldered a capillary containing kolibration in the defined concentration scale mix deuterium and non deuterium of dimethyl sulfoxide (DMSO), giving ²D a nuclear magnetic resonance a signal in the field of 3,4 m.p. (rather (ÑD₃)₄Si),while ²D a nuclear magnetic resonance signal HDO is in area of 4,7 m.p. (Rather (ÑD₃)₄Si).

Peaks of received signals are displayed in a picture 1.

Handling of the received spectra consisted in determination of a ratio integral intensity ²D a nuclear magnetic resonance of signal HDO, containing in the researched sample rather ²D a nuclear magnetic resonance of signal DMSO-D₁ which intensity, in turn, has been defined under the same conditions concerning standards – samples of water with precisely certain content of deuterium (3,7 ppm, 51 ppm, 150 ppm). Measurements of each sample were led repeatedly for reduction of errors of experiment. Thus accuracy of determination of the content of deuterium in biological samples is 2± ppm.

Picture 1.- Relation of the integral intensity ²D a nuclear magnetic resonance of signal HDO, rather ²D a nuclear magnetic resonance of signal DMSO-D₁

The measurement of spectra EPR spent at ambient temperature on spectrometer JES FA300 (JEOL, Japan) in a X-range. The measurement conditions: the microwave oven capacity 1 mW, frequency of microwave radiation of 9144 MHz, amplitude of high-pitched modulation 0,1 ìÒ. Samples measured in a quartz ampoule (5 mm), the weight of a hinge plate in a resonator zone is 0,0300 g. Concentration of the paramagnetic centres in samples defined by comparison with a standard sample signal (TEMPOL). Integral signal strength EPR in investigated samples defined by a double numerical integration on a method of rectangles [8].

In thirty days, after the beginning of carrying out of experiment in skilled and control groups watched exterior differences. The animals in skilled group outwardly looked healthy, without any deviations. In a check group watched organic violations on ears, to a nose. Biopsy research has shown that some rats in a check group had a formation of the Nevus of Bekker – pigmentary hair epidermal a nevus to an inflammation. CODE ÌÊB: CODE D23.3 Other good-quality new growths of a skin.

By this researches it is installed that indexes antioxide activity at animals in I group have made plasmas of blood 1287,3±11,6. And in animals of II group indexes ÀÎS were above on 9,2 % in comparing by group I (p <0,05) that testifies to increase at them activity ÀÎS and allows to speak about improving of system operation of nonspecific protection of an organism. At rats of II group indexes free radical oxidation have been lowered on 24,8 % in comparison with group I that specifies in reduction of processes of formation of the free radicals at usage of water with the lowered content of deuterium. Such changes prooxide-antioxide with prevalence antioxide potential at system level should be considered systems as the factor of increase of nonspecific protection of an organism.

The result of this research the content of deuterium in blood plasma is resulted in table ¹1.

Table ¹1 – The deuterium content in blood plasma of rats

Sample	Changes of deuterium content, %
Plasma of blood	15,2

Apparently from table 1, after the use by rats of water with the under content of deuterium within a month, its concentration in blood plasma has decreased on 15,2%.

Spectra EPR of samples of heart of laboratory animals contain a non-isotropic singlet signal, parametres of spin-gamiltoniana (g_^ = 2.0074, g_֕ = 2.003) correspond to stable radicals [9,10]. Spectra EPR of samples of a liver have similar character.

The results of measurement of concentration of paramagnetic centers (PÌC) in lyophilization fabrics of a liver and heart of laboratory animals, show that amount PMC in researched fabrics decreases approximately in 4 times in comparison with a check group that testifies to favorable influence of water with the under content of deuterium on an organism of animals.

In lyophilization fabrics of kidneys of differences between skilled and control groups it is not revealed.

By this way, it is installed that the use of water with the under content of deuterium concerning natural level HDO, leads to lowering of concentration of this element in blood plasma, erythrocytes and hearts of laboratory animals. Such changes in turn, conduct to reduction prooxide loadings in an organism and to balance recovery of prooxide-antioxide systems that further is accompanied by increase of immunity to the laboratory animals[11].

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