[REDUCING RESISTANCE TO ACID HEMOLYSIS BY IRON-CONTAINED DRUG INCREASES THE LEVEL OF HEMOGLOBIN IN THE ERYTHROCYTES OF AGING ANIMALS.]

In experiments in vivo we studied the effect of chronic iron-contained drug (Urfuhem) supplemention on the level of hemoglobin (Hb) in the blood of aging rats. To establish the biochemical mechanisms of drug action it were determined the parameters of oxidative/nitrosative stress and the hydrogei sulfide level in plasma and erythrocytes, the level of non-heme iron in plasma and erythrocytes sensitivity to acid hemolysis. It was found that in aging rats the drug significantly increases the Hb content of red blood cells and reduces its resistance to acid hemolysis. After the drug supplemention the rate of superoxide anion-radical (*0(2)(-)) generation in erythrocytes and stable hydrogen peroxide (H(2)0(2)) content both in plasma and erythrocytes, were down-regulated. The drug did not reduce the high levels of generation of the hydroxyl radical (*OH) and high levels of excess NO de novo synthesis by iNOS in erythrocytes but reduced the pools of nitrate anion(NO(3)(-))a its reutilization for NO synthesis. After thei drug supplemention the rate of constitutine NO synthesis by cNOS in aging rats plasma was up-regulated perhaps by cNOS coupling. The results indicate that the reason for increasing the permeability of the proton (H*) in red blood cells that causes the acid hemolysis in aging fats after the drug supplemention can be change in the balance of levels of oxidative and nitrosative stress in red blood cells in favor of the latter, and that toxic, OH generation is not at the expense of the classical Fenton reaction in the presence of iron ions (Fe(2+)); but due to the formation and decomposition of peroxynitrie (ON0O(-)).

INDUCTION OF OXIDATIVE AND NITROSATIVE STRESS IN BOYS IN ADAPTING TO PHYSICAL STRESS DURING TRAINING AND COMPETITIVE PERIODS.

We studied the features of development of oxidative and nitrosative stress in otherwise healthy individuals under the influence of prolonged exercise of high volume and intensity. It is shown that young men who systematically performed muscular work have a high content of markers of different ways of generation of superoxide radical, a reactive oxygen species for products of lipid peroxidation and markers of nitrosative stress. The increase in the degree of adverse effects on the body intensive training and competitive loads is accompanied by pronounced adaptive changes in the hierarchy of oxidizing constitutive de novo synthesis of nitric oxide, as well as its nonoxide reutilization synthesis (in 3 times higher). Disadaptation of the organism of boys at the end of the competition period is reflected in growing levels of generation of ROS (superoxide radical: 3,5 times higher, hydrogen peroxide: 2,5 times higher). The products of purine nucleotides degradation were 2 times higher, and the increase in the content of the nitrate anion was 2,5 times higher.

[PHYSICAL EXERCISE TRAINING CAN- CELS CONSTITUTIVE NOS UNCOUPLING AND INDUCED VIOLATIONS OF CARDIAC HEMODYNAMICS IN HYPERTENSION (PART III)].

In the heart and heart mitochondria spontaneously hypertensive rats investigated the effect of physical exercise training (swimming in a moderate and excessive training mode) on the physiological indicators of cardiac hemodynamics and biochemical parameters that characterize the level of oxidative and nitrosative stress. The index of coupling Ca(2+)-dependent constitutive NO-synthases (cNOS = eNOS + nNOS) and biochemical index of dysfunction were calculated. It turned out that both modes of training is completely restored, and even exceed the reference values in untrained rats Wistar conjugate cNOS state and Ca(2+)-dependent synthesis of nitric oxide (NO). Intensity regime of exercise on the border of functionality have been ineffective for improving the functional state of the cardiovascular system and hypertension can provoke it further. Moderate physical training regime, on the contrary, improves the diastolic function of the heart due to an increase dP/dtmin, reducing end-diastolic pressure and a significant reduction in end-diastolic stiffness. Moderate exercise decreased peripheral resistance and cardiac afterload, as indicated by the decrease in end-systolic pressure and arterial stiffness, which contributed to more efficient and energy-saving of heart work. Improve physiological indicators of cardiac hemodynamics and functional state of the heart in moderate mode of training correlated with changes in both the calculated indices. Moderate mode of training is recommended as a simple physiological preconditioning method for the prevention of cardiac dysfunction, hypertension as a result of state uncoupling cNOS and the resulting excessive generation of superoxide and, conversely, inhibition of Ca(2+)-dependent synthesis of NO.

[NOS UNCOUPLING IS ACCOMPANIED WITH INDUCTION OF THE OXIDATIVE STRESS AND THE CARDIOHEMODYNAMICS DISTURBANCES IN HYPERTENSION].

We compared the performance of cardiaohemodynamics and indicators of oxidative and nitrosative stress in the heart and aorta in normotensive Wistar rats (WKR) and spontaneously hypertensive rats (SHR). On the basis of experimentally determined parameters to calculate cNOS uncoupling index and biochemical index of function (BIF) in these organs of the cardiovascular system. In the heart, and especially in the aorta of SHR develop a combined oxidative and nitrosative stress that leads to cNOS uncoupling, BIF lowering that correlate with lowering of systolic and diastolic functions, inhibition of the efficiency Frank-Starling mechanism, oxygen consumption of the heart and increasing arterial stiffness. We made the assumption of the existence of the vicious circle of enhancing oxidative stress in organs of the cardiovascular system due to additional superoxide generation by uncoupling cNOS.

[ENDOTHELIAL MONOCYTEACTIVATING FACTOR II CANCELS OXIDATIVE STRESS, CONSTITUTIVE NOS UNCOUPLING AND INDUCED VIOLATIONS OF CARDIAC HEMODYNAMICS IN HYPERTENSION (PART II)].

The purpose of this study was to investigate the effect of EMAP II on free radical state of the heart and blood vessels, to restore cNOS coupling and cardiac hemodynamics in spontaneously hypertensive rats. It was found that, due to the combined inhibition of oxidative and nitrosative stress, EMAP I quickly restores impaired in hypertension constitutive de novo synthesis of NO by restoring cNOS coupling. Restoration by EMAP II of constitutive de novo synthesis NO abolished cardiac and endothelial dysfunction in spontaneously hypertensive rats. In hypertension, the introduction of EMAP II helped to improve the performance of the pumping function of the heart (stroke volume increased by 18.2 %, cardiac output -22 %), an arterial stiffness decreased by 23.2 %, process of relaxation of the left ventricle improved, due to decreased in 4,7 times myocardial end-diastolic stiffness.

[Nanocerium restores the erythrocytes stability to acid hemolysis by inhibition of oxygen and nitrogen reactive species in old rats].

In experiments in vivo the effect of nanocerium (cerium oxide nanoparticles) on the stability of red blood cells to acid hemolysis, levels of both ROS and RNS generation and H2S pools in plasma and erythrocytes of old rats were investigated. In red blood cells of old rats the proton penetration into the matrix of erythrocytes showed a significant raising and the fate of labile "aging" erythrocytes in old animals compared with adult were up- regulated. These phenomena paralleled with significant up-regulation of ROS and RNS generation. Introduction for 14 days per os to old rats 0.1 mg/kg of nanocerium fully restored resistance of erythrocytes to acid hemolysis by ROS and RNS in both plasma and erythrocytes reduction. Nanocerium decreased the erythrocytes and, conversely, significantly increased the plasma's pools of H2S.

[BRAIN FOCAL ISCHEMIA-REPERFUSION CAUSES A DECREASED RESISTANCE OF ERYTHROCYTES FROM VENOUS BLOOD TO ACID HEMOLYSIS, WHICH IS PREVENTED BY ECDYSTERONE].

We investigated the resistance of erythrocytes from rat brain venous blood to acid hemolysis in the dynamics of brain ischemic period (15, 30, 45 and 60 min), as well as in the early (5 min) and distant (24h) period of brain reperfusion. Brain ischemia-reperfusion was made in rats that received ecdysterone (standartized extract of Serratula coronata) within 18 days (per os, 1 mg/kg). Analysis of the kinetic curves of acid hemolysis showed a pronounced (60 times, from 1.45 to 85.85% at 60 min of brain ischemia and at 5 min of brain reperfusion, respectively) increase of unstable erythrocytes that hemolyzed easily (< 2.5 min). In the preconditioned rats, this increase was only 8-fold. During the period of brain ischemia, with a maximum at 15th minute, in the venous blood from brain the diene conjugates (DK) pools increased from 2.40 to 9.48 ng/mg protein and LTC4 pools increased from 1.49 to 5.98 pmol/mg protein. Even more pools of DC and LTC4 were increased at 5th min of brain reperfusion. In animals received ecdysterone, during ischemia and early reperfusion period, both pools of DC and LTC4 in venous blood were lower than that in the controls. The latter implies a possible antiradical mechanism of the protective effect of ecdysterone.

Some signal effects in human erythrocytes on specific binding of free doxorubicin and doxorubicin conjugates with magnetite nanoparticles.

Binding of doxorubicin (DOX) immobilized on nanodispersed magnetite (DOX-M conjugates with loading in the range of 0.16-25.1 mg DOX/g carrier) to intact human erythrocytes in concurrence with free DOX was investigated. Two specific binding sites for doxorubicin were revealed at the plasma membrane of human erythrocytes. Changes in the ordering of the DOX-M nanoparticles according to small angle scattering data are consistent with their specific binding at the plasma membrane upon incubation with erythrocytes. Free and conjugated doxorubicin modulated signal transduction in erythrocytes in a similar way. Both up-regulate nitric oxide and cyclic GMP and down-regulate cyclic AMP production and stabilize the membranes of oxidatively damaged erythrocytes.