• R. F. Aminov Zaporizhzhya National University
Keywords: hirudo influence, biologically active substances, proliferation


Nowadays, hirudotherapy, namely the treatment with medical leeches, is re-gaining momentum in the use with the prophylactic and therapeutic goal, because there are practically no diseases that can not be treated with leeches. Of the more than 200 kinds of leeches that live on the planet, only three types of medical leeches are most often used in treatment. Currently, medical leeches are grown on special biopharmic plants, after which they undergo surgical medical control. Leech is a single tool which eliminates patient infection diseases transmitted through blood. Hirudotherapy is a method of treatment that has no analogues in modern medicine. So, as now in the pharmacology more and more are shown by side effects from their use in the treatment of a number of diseases. Researchers are increasingly beginning to study and explore various natural-therapeutic methods of influencing the body of an animal and humans, one of which is hirudotherapy. In the saliva of a medical leech contain more than 100 substances that have many therapeutic effects. In the saliva of a medical leech contain more than 100 substances that have many therapeutic effects. Hirudotherapy and peptic salivary drugs are used effectively in medicine and in veterinary medicine. The main side effects of drugs are their negative effects on the proliferation of blood cells. Therefore, it is important to investigate the proliferative activity of lymphocytes by means of the reaction of blast transformation of lymphocytes in the whole blood of not only mature female rats, but also in their offspring against the background of hirudotherapy. Adult sexually mature female non-linear rats weighing 150-200 grams, two weeks prior to pairing and two weeks after, were prepared by one hungry medical leech Hirudo verbana (Carena, 1820), each week weighing 400 mg (4 consoles). Fixed animals using a fixing device. Animals were divided into two groups: the first experimental group of animals for the influence of biologically active substances Hirudo verbana; The second control group of animals without interference. The females were studied after feeding the offspring, as well as the growth in the dynamics at 45 and 60 days.

The proliferative activity of blood lymphocytes was evaluated in the whole blood by the reaction of blast transfection of lymphocytes (RBTL). Blood dissolved in 350 μl of heparin and adjusted to 2,5 ml of medium (1: 5): 50 ml of medium 199, 5 ml of 10 % embryonal calf serum, 15 mg of glutamine, 119 mg of Hepes, 50 μg / ml of gentamicin, 0,05 mM of 2 -mercaptoethanol. The mixture was poured into penicillin vials and saturated with CO2 pipettes. The culture mixture was introduced by mitogen – konkanavalin A (KonA, Germany) at a dose of 10 μg / ml. As antigens (AG), salt extract was used from bodies N. verbana Carena, 1820 at a dose of 5 μg / ml. The concentration of protein in the extract was determined by the Lowry method (Menshikov, 1987). The mitogenic stimulation of lymphocytes by plant lectin ConA was carried out to determine the conditionality of cell culture conditions, as well as to study the potential proliferative activity of the major populations of T-lymphocytes. The vials were sealed and cultured for 24 hours at a temperature of +37°. After 24 hours of cultivation, the samples were centrifuged (5 min at 1500 rpm), the culture supernatant was selected, and from the cell sieve, preparations were prepared, fixed with methanol, stained with Romanovsky-Gimza, followed by differentiation in acidified hydrochloric acid with distilled water. The level of RBTL was evaluated morphologically, taking into account 300 - 400 lymphocytes from the beginning of the brush to the basis of the drug. Statistical processing of data was carried out using the SPSS v.21,0 computer program. (IBM SPSS Statistics., USA). The sample parameters given in the table below have the following notation: X is the sample mean, SE is the standard error of the mean. The probability of differences between the average values was estimated according to the Student criterion. The differences were considered reliable at р < 0,05.

As a result of the study in mature females, scurries of the control and experimental groups of animals, when stimulated by plant mitogen and leech antigens, the percentage of blast cells increases p < 0,05 Comparing experimental with control groups of spontaneous cultures and stimulating vegetative mitogen almost did not differ from each other, with some the tendency to increase in the research group. And when stimulated by the AG, the experimental group significantly exceeded the figures in comparison with the control group, both in females and in their offspring, p < 0,05.

Blasters stimulated with plant lectin were typical, indicating a productive lymphogenesis. When stimulated by antigens, blastus had an underdeveloped cytoplasm and reduced its basophility, as a result of insufficient development of the protein-synthetic system.


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