Rat peripheral blood leukocyte subset composition, hemogram, lymphoid organ and body morphometry after exposure to biologically active substances derived from Hirudo verbana saliva

Medical community has noted that the prevalence of allergic diseases by steadily tends to rise resulting from using synthetic drugs ecological catastrophe. In this regard, more and more attention is increasingly being paid to natural therapeutic approaches, one of which is based on applying hirudotherapy. There were used 40 pubescent outbred female rats and their 200 pups born during experiment. In particular, female rats underwent leeching in the subscapular region 2 weeks before and 2 weeks after mating by applying medical leeches weighing 400 ± 10 mg. Next, relevant progeny was examined at day 1, 15, 30, 45, and 60 after birth, whereas female rats– after weaning. Experiment was designed to assess diverse parameters in accordance with the age-related rat development: newborn pups – 1-5 days old, day 6-21 – suckling period, day 22-50 – onset of puberty, day 60 onward – puberty. After measuring body morphometric parameters, female rats were sacrificed by decapitation under ether narcosis on day 60, whereas rat pups – at various time points. followed by autopsy and collecting blood sample into a sterile centrifuge tube by using 2% Spofa crystalline heparin (8:1) to be further examined by standard assays: morphometry of rat body, spleen and thymus; measuring total leukocyte count; CBC with differential; RBC; hemoglobin level; color index. Statistical data processing was performed by calculating arithmetic mean, error of arithmetic mean, standard deviation by using SPSS version 21.0 and Microsoft Office Excel 2010 software. Significance of differences was estimated by using Student's t test, and set at p≤0.05. Our study demonstrated medical leech saliva components applied during hirudolotherapy exerted an immune-augmenting effect of on morphometric body, thymus and spleen parameters. Moreover, starting from day 1, total leukocyte count, RBC, and hemoglobin level were increased in the experimental group. Moreover, we convincingly demonstrated a stimulatory effect of medical leech saliva components on histogenetic reactions at various ontogenetic stages in both female rats and their progeny. In the former, such morphogenetic effect was maintained for more than 70 days after the last leeching: on week 2 after mating and 30 days after delivery. Changes in morphological and hematological evidence about enhanced morphogenetic function of immune system in response to saliva-derived biologically active substances released by medicinal leeches.

hirudotherapy, medical leech, biologically active substances, morphometric indices of the body