Proliferative activity of bone marrow cells of rats under the influence of biologically active substances of a medical leech

Keywords: hirudotherapy, hirudology, stages of mitosis, leukocytes, erythrocyte, hemoglobin

Abstract

Hirudotherapy has been used in medicine since time immemorial. The advantage of this type of treatment is that the bite of a leech has a restorative effect on the entire body. The leech bites the skin and injects into the patient’s blood biologically active substances that activate blood microcirculation and promote immunity. The saliva of the leech contains more than 100 substances that contribute to the spectrum of therapeutic effects. But there are no studies on the effect of hirudotherapy on the proliferation of bone marrow cells, which is ontogenetically primary in the development of myeloid and lymphoid tissues. Therefore, it is important to study the mitotic activity of bone marrow of rats at the early stages of postembryonic development against the background of influence by the leech. Female nonlinear rats, two weeks before and two weeks after mating with males, were presented each week to one hungry medical leech weighing 400 mg. The females were examined after feeding their offspring and their offspring were likewise examined on day 1, 15, 30, 45, 60 of the experiment. The animals were kept in vivarium conditions on a standard diet in individual cages. All animals were decapitated under ether anesthesia and the number of leukocytes, the leukocyte blood count, the number of erythrocytes, hemoglobin, and the color index were examined. After that, the femurs were quickly removed and the adjacent muscles separated with a sterile gauze. We cut the epiphyses and cut along the bone. Bone marrow from the femur was flushed with a warm (37 °C) hypotonic 0.9% sodium citrate solution into a sterile centrifuge tube using a Pasteur pipette. The resulting cell suspension was incubated in a hypotonic solution for 10 minutes at 37 °C. The resulting suspension was centrifuged for 5 minutes at 1000 rpm leaving only a small residue of sediment in the test tube. The cells were fixed in methyl alcohol with glacial acetic acid (three parts methyl alcohol and one part acetic acid). 3,000 cells were analyzed, among which we determined those that are in mitosis, and expressed their share. As a result of our studies, the immunomodulating effect of the saliva of the medical leech was revealed, which manifested itself in an increase in the proliferative activity of bone marrow cells, which led to an increase in the number of red blood cells, increased hemoglobin and increase in the number of leukocytes. The color index and the leukocyte blood count were within the normal limits. 

References

Abdualkader, A. M., Ghawi, A. M., Alaama, M., Awang, M., & Merzouk, A. (2013). Leech therapeutic applications. Indian Journal of Pharmaceutical Sciences, 75(2), 127–137.

Abdullah, S., Dar, M. L., Rashid, A., & Tewari, A. (2012). Hirudotherapy. Leech therapy: Applications and indications in surgery. Archives of Clinical and Experimental Surgery, 1(3), 172–180.

Aminov, R. F., & Frolov, O. K. (2015). Vplyv biologichno aktyvnyh rechovyn sol’ovogo ekstraktu medychnoi’ p’javky na fagocytarnu aktyvnist’ nejtrofiliv i cytomorfometrychni zminy limfocytiv krovi ljudyny u kul’turi [Influence of biologically active substances in the salt extract of the medicinal leech on phagocytic activity of neutrophils and cytomorphometric changes of blood lymphocytes in culture]. Naukovyj Visnyk Chernivec’kogo Universytetu. Biologija, 7(1), 108–112 (in Ukrainian).

Aminov, R. F., & Frolov, O. K. (2016). Fagocytarna ta metabolichna aktyvnist’ nejtrofiliv shhuriv na rannih etapah postembrional’nogo rozvytku za vplyvu biologichno aktyvnyh rechovyn sol’ovogo ekstraktu Hirudo verbana [Phagocytic and metabolic activity of neutrophils of rats in the early stages of post-embryonic development under the influence of biologically active substances of the salt extract of Hirudo verbana]. Visnyk of Dnipropetrovsk University. Biology, Medicine, 7(2), 96–100 (in Ukrainian).

Aminov, R. F., Frolov, A. K., & Fedotov, E. R. (2016). The impact of fetal load of biologically active substances of Hirudo verbana saline extract antigens on the morphometric and hematologic parameters of rats in the early stages of post-embryonic development. Immunopathology, Allergology, Infectology, 4, 6–11.

Bank, J., Zilinsky, Y., Haik, J., Winkler, E., & Goldan, O. (2008). Medicinal leech fixation in precarious locations. Reconstructive Microsurgery, 24, 67–68.

Berkalo, L. V. (2003). Metody klinichnyh ta eksperymental’nyh doslidzhen’ v medycyni [Methods of clinic eksperimentalnih dozen in charity fund medical]. Polimet, Poltava (in Ukrainian).

Bogdanova, M. S. (2016). Kletochnye kul’tury [Cell cultures]. Politehn, Saint Petersburg (in Russian).

Canpolat, İ., & Sağlam, N. (2004). Treatment of diffuse hematoma in a dog with the medicinal leech, Hirudo medicinalis. Doğu Anadolu Bölgesi Araştırmaları, 2(2), 97–99.

Chepeha, D. B., Nussenbaum, B., Bradford, C. R., & Teknos, T. N. (2002). Leech therapy for patients with surgically unsalvageable venous obstruction after revascularized free tissue transfer. Archives of Otolaryngology Head and Neck Surgery, 128, 960–965.

Demidchik, L. G. (2001). Girudoterapija pri skrytom mastite korov [Hirudotherapy with hidden cow mastitis]. Veterinarija, 2, 441 (in Russian).

Eldor, A., Orevi, M., & Rigbi, M. (1996). The role of the leech in medical therapeutics. Blood Reviews, 10, 201–209.

Flaum, R. A., & Shahov, A. V. (2008). Primenenie girudorefleksoterapii v lechenii pacientov s zabolevanijami vnutrennego uha [Application girudorefleksoterapii in the treatment of patients with diseases of the inner ear]. Medicinskij Al’manah, 2, 63–65 (in Russian).

Frodel, J., Barth, P., & Wagner, J. (2004). Salvage of partial facial soft tissue avulsions with medicinal leeches. Archives of Otolaryngology Head and Neck Surgery, 131, 934–939.

Frolov, A. K., & Litvinenko, R. A. (2014). Reaction of patients’ lymphocytes’ blast-transformation in blood, stimulated by plant mitogens and annelid antigens at hirudotherapy. Immunopatologija, Allergologija, Infektologija, 3, 10–13.

Frolov, A. K., & Litvinenko, R. A. (2015). Basic morphofunctional features of pharmaceutic leech (Hirudo verbana Carena, 1820) tissues in various forms of response after hirudotherapeutic procedures. Annals of Parasitology, 61(1), 27–35.

Frolov, A. K., Kopejka, V. V., & Fedotov, E. R. (2010). Vlijanie girudoterapii na fiziologicheskie pokazateli u koz [The impact of treatment on physiological parameters in goats]. Zhivotnovodstvo Ukrainy, 7, 7–10 (in Russian).

Hildebrandt, J. P., & Lemke, S. (2011). Small bite, large impactsaliva and salivary molecules in the medicinal leech, Hirudo medicinalis. Naturwissenschaften, 98(12), 995–1008.

Hullett, J. S., Spinnato, G. G., & Zi, V. (2007). Treatment of an ear laceration with adjunctive leech therapy: A case report. Journal of Oral and Maxillofacial Surgery, 65, 2112–2114.

Isahanjan, G. S., & Arutjunjan, V. M. (1991). Medicinskie pijavki ih lechebnoe primenenie v terapevticheskoj klinike [Medical leeches: their therapeutic use in a therapeutic clinic]. Terapevticheskij Arhiv, 63(8), 110–112 (in Russian).

Kamenev, O. J., & Baranovskij, A. J. (2010). Lechenie pijavkami – teorija i praktika girudoterapii [Treatment by leeches: Theory and practice hirudotherapy]. Ves’, Saint Petersburg (in Russian).

Koeppen, D., Aurich, M., & Rampp, T. (2014). Medicinal leech therapy in pain syndromes: A narrative review. Wiener Medizinische Wochenschrift, 164(5–6), 95–102.

Krjachko, O. V., & Lukojanova, L. A. (2009). Vlijanie girudoterapii na nekotorye gematologicheskie i biohimicheskie pokazateli pri intoksikacionnom sindrome u sobak [The influence of hirudotherapy on some hematologic and biochemical indices in case of intoxication syndrome in dogs]. Veterinarnaja Praktika, 1, 65–68 (in Russian).

Kumar, S. A. (2012). Anti inflammatory effect of leech therapy in the patients of psoriasis. Journal of Pharmaceutical and Scientific Innovation, 1(1), 71–74.

Matvjejeva O. V., Viktorov O. P., Blihar V. J., Puhlyk B. M., Chop’jak, V. V., Jajchenja, V. P., Logvina, I. O., & Bykanova, I. I. (2011). Do pytannja klasyfikacii’ pobichnyh reakcij likars’kyh zasobiv ta pidhodiv do i’h dyferenciacii’ (bibliografichnyj ogljad) [To the question of classification of adverse reactions of medicinal products and approaches to their differentiation (bibliographic review)]. Ukrai’ns’kyj Medychnyj Chasopys, 82, 78–84 (in Ukrainian).

Meriaux, C., Arafah, K., Tasiemski, A., Wisztorski, M., Bruand, J., Boidin-Wichlacz, C., Desmons, A., Debois, D., Laprévote, O., Brunelle, A., Gaasterland, T., Macagno, E., Fournier, I., & Salzet, M. (2011). Multiple сhanges in рeptide and lipid expression associated with regeneration in the nervous system of the medicinal leech. PLoS ONE, 6(4), e18359.

Michalsen, A., Lüdtke, R., Cesur, O., Afra, D., Musial, F., Baecker, M., Fink, M., & Dobos, G. J. (2008). Effectiveness of leech therapy in women with symptomatic arthrosis of the first carpometacarpal joint: А randomized controlled trial. Pain, 137(2), 452–459.

Mineo, M., Jolley, T., & Rodriguez, G. (2004). Leech therapy in penile replantation: A case of recurrent penile selfamputation. Urology, 63, 981–983.

Mumcuoglu, K. Y. (2014). Recommendations for the use of leeches in reconstructive plastic surgery. Evidence-Based Complementary and Alternative Medicine, 1, 1–7.

Popovа, L. K., & Petrov, A. N. (1999). Girudoakupunktura pri subklinicheskom mastite u korov [Hirurdoacupuncture with subclinical mastitis in cows]. Veterinarija, 10, 23–24 (in Russian).

Pospelova, M. L., & Barnaulov, O. D. (2010). Effects of hirudotherapy on intravascular thrombosis activation in different groups of patients with cerebrovascular pathologies. Aktuelnosti iz Neurologije, Psihijatrije i Graničnih Područja, 18(3), 27–32.

Romanova, E. M., Klimina, O. M., & Kozlova, L. A. (2008). Primenenie girudoterapii i girudoakupunktury pri subklinicheskoj forme mastita u korov [The use of hirudotherapy and hirudoacupuncture in the subclinical form of mastitis in cows]. Veterinarnyj Vrach, 4, 35–37 (in Russian).

Savinov, V. A. (2004). Girudoterapija [Hirudotherapy]. Medicina, Moscow (in Russian).

Sobczak, N., & Kantyka, M. (2014). Hirudotherapy in veterinary medicine. Annals of Parasitology, 60(2), 89–92.

Zaidi, S. M., Jameel, S. S., Zaman, F., Jilani, S., Sultana, A., & Khan, S. A. (2011). A systematic overview of the medicinal importance of sanguivorous leeches. Alternative Medicine Review, 16(1), 59–65.

Zapadnjuk, I. P., Zapadnjuk, V. I., & Zaharija, E. A. (1983). Laboratornye zhivotnye. Razvedenie, soderzhanie, ispol’zovanie v jeksperimente [Laboratory animals. Breeding, keeping, use in the experiment]. Vishha Shkola, Kiev (in Russian).

Zharov, D. G. (2003). Sekrety girudoterapii ili kak lechit’sja pijavkami [Secrets of treatment or how to treat leeches]. Feniks, Rostov (in Russian).

Published
2017-11-11
How to Cite
Aminov, R. F., & Frolov, A. K. (2017). Proliferative activity of bone marrow cells of rats under the influence of biologically active substances of a medical leech. Regulatory Mechanisms in Biosystems, 8(4), 501–505. https://doi.org/10.15421/021777