Liver plays an important role in homeostasis maintenance. The organ takes part in its maintenance both in normal processes supporting a life, and in adaptation of an organism to any loadings and stresses. The morphological and functional condition of this organ can indirectly reflect a condition of an organism as a whole [5]. Any change of such parametres as the linear sizes of a cell nucleus and a number of hepatocytes is one of typical signs of adaptation process taking place as the sizes of a liver cell nucleus directly depend on the amount of DNA in it [7]. The excitation or damage condition of hepatocytes can be accompanied by more intensive protein synthesis. The same phenomenon is also observed at the increase of functional activity of an organism as a whole. Moreover, it is possible to estimate a functional condition of body according to the linear sizes of hepatocytes´ nucleus. Besides, it´s typical of a liver to have rather constant correlation of diploid, tetraploid and polyploid hepatocytes. Any change of this correlation shows some functional disorder in the organ which can be caused by various damaging factors. The total number of cells and a correlation of various cellular populations in a normal condition is supported by the interaction of mitosis, amitotic division and apoptosis mechanisms [1].

One of ways to increase the adaptation abilities of separate organs, their systems and those of an organism as a whole is to apply biologically active additives (BAA). BAA´s supply (manufacture) is an important branch of modern biotechnology [3,4].

BAAs produced from dead bees take an important place among other additives made from other products of beekeeping. Dead bees continue to possess high biological activity that results from the components building their structure such as: chitin, melanin, heparoids and other biologically active substances (BAS). These preparations (BAAs) possess adaptogenical (adaptation-stimulating), immunomodulatory, hepatoprotective, recycling and antitoxic properties [6].

Proceeding from the above-stated, the research of the influence of an extract from dead bees on a liver during a chronic or sharp stress seemed to us quite promising.

Vistar rats, both male and female, at the age of 1,5 months were chosen to make a set of experiments. The first group of animals (n=20) was a control one, the second group (n=20) was an experimental one, in the latter group all rats received an extract of dead bees with a drink, its amount being calculated according to a rat´s weight, i.e. 0,1 g of an extract = 100 g of weight. Both groups were subdivided into 2 subgroups. All the animals were kept in the standard laboratory conditions.

Within three weeks a chronic stress was provided in one of the subgroups of each group. Two other subgroups were subject to an immobilization stress. A chronic stress consisted in using a test of forced swimming which was carried out by means of the standard technique [2]. The rats of a control and experimental groups were forced to swim until complete exhaustion once a week throughout 3 weeks, after that the sampling of liver cells for morphological and histochemical analysis was taken. The experiments on animals were carried on according to the regulations of protection of the vertebrate animals used in the scientific purposes (Regulations and recommendations for the European independent committees concerning ethics, Brussels, 1995, 1997; Recommendations to Committees on the ethics, making examination of biomedical research, Geneva, 2000).

An immobilization stress was simulated by fixing rats, with their paunches up, on a laboratory restraining chair for an hour, then the animals were put down.

Histologic research was carried out by the standard technique with samples dyed by hematoxylin and eosin, with the subsequent calculation of mitotical and apoptotical indices (MI and AI) being made. These indices were calculated according to a number of hepatocytes under mitosis or apoptosis conditions in 1000 cells.

As a result, it has been established that the signs of the initial stage of a granular dystrophy were observed in the liver of all rats in the control group. The micronecrosis process was registered. The hepatocyte borders were blurred, they could not be accurately distinguished. The joist (beam) structure was damaged in some places. The morphological analyses result of rats´ liver in the experimental groups corresponded to the standard results under normal conditions.

MI for the rats in the control group subject to a chronic stress appeared to be the least one (1,175±
0,08 ‰) and evidently differed from the result (index) in the experimental group (2,00±0,1 ‰) and from MI of the immobilized rats´ liver in the control group (2,23±0,12 ‰).

The similar tendency has been traced as far as AI is concerned: in the control group at a chronic stress AI was equal to 0,025±0,001 ‰; in the experimental group under a chronic stress it amounted to 0,04±0,005 ‰ that evidently differed from the previous result. In the control group subject to a sharp stress AI was equal to 0,2±0,06 ‰ that appeared to doubtfully be higher than in the experimental group - 0,17±0,03 ‰.

Karyogram of rats´ hepatocytes in both control groups has clearly shown 3 peaks with the displacement tendency to the right that is characteristic for stress reactions which is accompanied by the increase of hepatocytes. In a liver of the experimental rats the karyogram had a more smoothed character, with 2 peaks being distinguished without any doubt. In a whole, the karyogram was displaced to the left. It is necessary to mention also that the karyogram for both groups which were under the conditions of a chronic stress was displaced to the left as compared to the immobilized rats´ karyogram.

The spirit extract produced from dead bees stimulates the process of regeneration of hepatocytes population by means of a mitosis under both chronic and sharp stresses. In addition, AI in liver of the rats that were given the dead bees extract proves to be higher alongside a better pathomorphological result. This can be explained by the fact that under some stressful factors hepatocytes originally perish due to apoptosis, and only after that one can see both monocelluar and local necrosis in the organ[8]. Taking into account the above, we can come to the following conclusion that the spirit extract produced from dead bees possesses a certain activity stimulating stress adaptation processes in the liver.

References

1. Areshidze D.A. Energoinformational condition of a liver of a human in past middle age at some pathologies and pathological conditions. Bulletin MSRU. №1 - 2009 - p. 89-93.
2. Bobkov J.G., Vinogradov V. M, Katkov V.F. Pharmacological correction of an exhaustion. - М: Medicine, 1984 - 207 p.
3. Krylov V. N, Smirnov A.M., Plotkin E.V., Ulitin I.B. Apilak and Apingalin in treatment of lungs sick of chronic nonspecific diseases. Thesis of a report of VI Union Russian conference of apiotherapy. 14-17 October 1997. Ryazan, 1998. p.161-164.
4. Kryzhanovskaya E.V. Biologically active substances in veterinary science. Author´s dissertation abstract on competition of degree of a Dr. Biological Science. - Shchelkovo, 2008. - 49 p.
5. Podymova S.D. Illnesses of a liver. - М: Medicine, 2005. - 768 p.
6. Rashidova S.S., Aripova T.U., Voropaeva N.L., Milusheva R. U, Pulatova S.R., Ruban I.N. Immunomodulating activity of a chitosan polymeric metallocomplexes. // Materials of a VIII international conference «Modern prospects of chitin and chitosan usage ». - Kazan, 2006
7. Tashke K. Entrance in quantitative cyto-histologic morphology. - Bucharest.: Publishing house of Academy of Socialist Republic of Romania, 1980.-191 p.
8. Filchenkov A.A., Stoika R.S. Apoptosis and a cancer. - K.:Morion, 1999. - 184 p.

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The work was submitted to international scientific conference «Prospects for the development of university science», Dagomys (Sochi), September 21-24, 2009, came to the editorial office on 20.07.2009.