[Ultradian rhythms and oxidative stress in the tissue of the lymph nodes in ontogenesis.]

We studied the age-related features of the ultradian rhythms of levels of total protein and albumin in the lymph nodes under normal condition and chronic toxic oxidative stress condition using the chronobiological approach and the biochemical methods. During the experiment, we compared the rhythmic activity of levels of total protein and albumin in the rat lymph nodes in ontogenesis and during chronic exposure to sulfur-containing natural gas. The revealed fluctuations of the levels of total protein and albumin can be characterized as around-hourly (ultradian) rhythms, which period is approximately 20-40 to 60 minutes. The obtained experimental data indicate a decrease in the adaptive resistance of lymph node with age and the depletion of the antioxidant system of rats in aging. These results could have a practical value for the development of correction methods of negative effects of toxic oxidative stress in ontogenesis.

[Cytoflavin effect on locomotor and psychoemotional status in physiological and pathological aging.]

Currently, the actual problem is the correction of motor, cognitive and psychoemotional disorders in physiological aging, as well as in various pathological processes that accompany aging and accelerate it. In this regard, it became necessary to search for drugs that can restore age-related disorders of the brain. The aim of the study was to evaluate the possibility of Cytoflavin as a pharmacological corrector of age-dependent disorders of the functions of the cerebral cortex during physiological and pathological, accelerated aging. The mouse sensorimotor cortex of the brain was the material for study. The transgenic male mice with HER2/neu overexpression at the age of 2 and 10 months were used as an experimental model, male wild-type FBV/N mice at the age of 2 and 18 months served as a control. We studied locomotor activity, orientational research behavior and the psychoemotional status of animals using the «open field¼ test and the Suok test. It was found that in old FBV/N mice, after the cytoflavin treatment, recovery of locomotor functions and orientational-research behavior is observed. Under conditions of HER2/neu overexpression after the Cytoflavin treatment, an improvement in motor functions occurs. It was also shown that the studied drug has an anxiolytic effect on both wild-type FBV/N mice and transgenic HER2/neu mice during aging. Thus, the positive effect of Cytoflavin on the dynamics of the behavior of experimental mice during physiological and pathological accelerated aging allow to suggest that in the late stages of ontogenesis, Cytoflavin restores the cerebral cortex functions and prevents neurodegeneration.

[Pharmacological correction of alterations of apoptosis of the neurons of the hypothalamus suprachiasmatic nucleus and pinealocytes during aging and stress.]

As is known, the pineal gland plays an important role in adaptogenesis, and the hypothalamus is one of the main links of the stress-reactive system and is involved in the regulation of the involution of the whole organism. So, the study of changes in these organs during stress and aging is very interesting. The aim of the work is to study the mechanisms of apoptosis of pinealocytes and neurosecretory cells of the suprachiasmatic nucleus of the hypothalamus during aging, stress, and under the conditions of pharmacological correction of involutional processes and stress response (antioxidant alpha-tocopherol acetate, immunomodulator cycloferon). We used Wistar rats as model, young (2-4 months) and old (30 months). Age-related features of the apoptosis dynamics of pinealocytes and neurosecretory cells of the hypothalamic suprachiasmatic nucleus were studied using TUNEL and immunohistochemistry, and the possibilities of pharmacological correction of apoptotic processes are determined. An age-dependent increase of apoptosis level of cells of suprachiasmatic nucleus and epiphysis in rats was revealed. The stress effect (immobilization) led to the intensification of cell death, more significant in older animals. The pineal gland and suprachiasmatic nucleus, traditionally regarded as regulators of circadian rhythms, are at the same time actively involved in general adaptation processes. The studied drugs (α-tocopherol-acetate, cycloferon, and their combination) have a pronounced anti-apoptotic, cytoprotective effect under physiological conditions during aging, as well as during non-specific emotional stress (immobilization) in young and old animals. The regulatory effect is accomplished by activating the expression of the anti-apoptotic protein Bcl-2 in the neurosecretory cells of the suprachiasmatic nucleus and pinealocytes.

[PHARMACOLOGICAL CORRECTION OF APOPTOSIS LEVEL OF CORTICAL NEURONS IN AGED HER2/NEU TRANSGENIC MICE].

Neurodegenerative changes and neuronal death are the basis for development of the nervous system aging. We investigated the mechanism of apoptosis of the sensorimotor cortex neurons of transgenic mice HER2/neu during aging, changes in the cortex function and the participation of exogenous neurometabolites (cytoflavin, piracetam) in regulation of neuronal death and locomotor and psycho-emotional status of mice. The level of apoptosis and expression of apoptosis markers (TUNEL, immunohistochemistry, Western blotting) in HER2/neu transgenic mice as compared to wild type mice (FBV line) were determined. In aging FBV mice the basal activity was shown to decrease and anxiety to increase correlating with the high level of neuronal apoptosis. We identified behavioral characteristics of transgenic HER2/neu mice and found that their low basal activity does not change with aging. Previously we have shown that in this strain of mice the apoptosis level is low, without any age-related changes, due to the suppression, first of all, of the p53-dependent pathway by HER2 (tyrosine kinase receptor) overexpression. Cytoflavin and piracetam were revealed to possess a marked neuroprotective effect, preserving and restoring functions of the nervous system (improving locomotion and psychological status) in both strains of mice. The effect of neurometabolites studied on neuronal apoptosis is ambiguous. In case of its low level it is a moderate stumulation of apoptosis via the external p53-dependent pathways with activation of caspase-3 in transgenic HER2/neu mice with high carcinogenesis level that can possibly prevent tumor development. On the contrary, in old wild-type animals we observed a significant decrease of age-dependent apoptosis level (by stimulating expression of the anti-apoptotic protein Mcl-1), which prevents neurodegeneration.