Aliev, Oleg I.

Plotnikov, Mark B.

Vasilev, A. S.

Zibareva, L. N.

English

Tomsk State University Repository

6890007 -4888/15/15950689  © 2015  Springer Science+Business Media New YorkBulletin of Experimental Biology and Medicine, Vol. 159, No. 5, September, 2015METHODSBrattleboro Rats as the Model of Blood Hyperviscosity Syndrome for Testing Substances with Hemorheological ActivityM. B. Plotnikov*,**, A. S. Vasil’ev*, O. I. Aliev*,**, and L. N. Zibareva**Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 159, No. 5, pp. 653-655, May, 2015Original article submitted June 17, 2014Hyperviscosity syndrome was described in Brattleboro rats. The aim of this study was to in-vestigate the possibility of Brattleboro rats using, as a test system for the study of agents with hemorheological activity. Under conditions of this model of high blood viscosity syndrome in Brattleboro rats, Lychnis chalcedonica L. extract (150 mg/kg) administered intragastri-cally for 10 days exhibited hemorheological activity by modulating macro- (plasma viscosity, fi brinogen concentration) and microrheological (erythrocyte aggregation and deformability) parameters. Hence, Brattleboro rats are an adequate model of hyperviscosity syndrome that can be used for search and testing of substances with hemorheological activity.Key Words: hereditary hypothalamic diabetes insipidus; Brattleboro rats; Lychnis chalce-donica extract; high blood viscosity syndrome; erythrocyte aggregation and deformability*E. D. Goldberg Research Institute of Pharmacology and Regen-erative Medicine; **National Research Tomsk State University, Tomsk, Russia. Address for correspondence: mbp2001@mail.ru. M. B. PlotnikovBrattleboro rats reproduce the main pathogenetic fac-tors of hereditary hypothalamic diabetes insipidus. In Brattleboro rats, signifi cant and pronounced shifts in macro- and microrheological parameters were found compared to Wistar rats: high values of whole blood and plasma viscosity, erythrocyte aggregation, fi-brinogen concentration, hematocrit, and erythrocyte deformability low [6]. Analysis of the hemorheologi-cal profi le attests to the formation of a pathological condition, high blood viscosity syndrome (HBVS) in Brattleboro rats.The aim of this work was to evaluate the possibility of using Brattleboro rats as the test systems for evalua-tion of substances with hemorheological activity.MATERIALS AND METHODSExperiments were carried out on 2.5-3.0-month-old male Brattleboro rats (n=16) and male Wistar rats (n=8) weighing 200-250 g. Brattleboro rats were ob-tained from the Research Institute of Cytology and Genetics, Siberian Division of Russian Academy of Sciences, and Wistar rats from the Department of Ex-perimental Biological Models, E. D. Goldberg Re-search Institute of Pharmacology and Regenerative Medicine. The experimental animals are conventional by their microbiological status.The rats were kept in standard Velaz plastic cages (5 per cage) on a bed of fi ne wood shavings under open regimen and on standard vivarium ration. The animals were kept in accordance with European Convention for the Protection of Vertebrate Animals used for Experi-mental and other Scientifi c Purposes (Strasburg, 1986). The air temperature in the vivarium was 20-23oC, hu-DOI 10.1007/s10517-015-3048-9690midity did not exceed 50%, air exchange 8:10 outfl ow-to-infl ow, light conditions (day:night) 1:1.After a 14-day quarantine, Brattleboro rats were divided into experimental and control groups. Dry Lychnis chalcedonica extract (LCE) exhibiting distinct hemorheological activity in various in vitro and in vivo models of blood hyperviscosity [1-4] was obtained from the aerial parts of Lychnis chalcedonica L. by repercolation with 40% ethanol solution, followed by lyophilization. LCE was standardized by ecdysterone content (0.74%). Experimental animals intragastrically received LCE suspension in 1% starch mucus in a dose of 150 mg/kg for 10 days; control rats received the same volume of 1% starch mucus. At the end of experiment, the rats were anesthe-tized with ether, blood samples were taken through a Tefl on catheter introduced into the common carotid artery, stabilized with 3.8% sodium citrate (1:9), and used for measuring whole blood viscosity, plasma viscosity, hematocrit, erythrocyte aggregation, and erythrocyte deformability. The viscosity of the whole blood and plasma were recorded on a rotary viscom-eter AKR-2. Hematocrit was determined by centrifu-gation in glass capillaries in a MHz-8 centrifuge. Fibrinogen content in the plasma was determined on a KG-IV coagulometer (Cormay) by Clauss chrono-metric method using a Tekh-Fibrinogen-test diag-nostic kit (Technologiya-standart). Erythrocyte ag-gregation and deformability were measured using a RheoScan AnD-300 instrument (RheoMeditech Inc.). Aggregation halftime served as the measure of aggre-gation activity of erythrocytes (the decrease in this parameter corresponded to an increase in erythrocyte aggregation capacity). Erythrocyte deformability was evaluated by the elongation index of erythrocytes (EIE) at a shear stress of 20 Pa. Oxygen availability for tissues was calculated by the hematocrit/viscosity ratio at 300 sec–1 [8]. The animals were sacrifi ced in a CO2-chamber.The data were processed statistically using Stu-dent’s t test.RESULTSIn Brattleboro rats, the whole blood viscosity in the entire range of shear rates was higher that in Wistar rats by 25-53% (Fig. 1), plasma viscosity by 20%, the concentration of fi brinogen by 51%, hematocrit by 7%, erythrocyte aggregation by 38%, and erythrocyte deformability by 21% (Table 1). Thus, the hemorheo-logical profi le in Brattleboro rats differed signifi cantly from that in Wistar rats. Based on the analysis of cer-tain hemorheological parameters and correlations be-tween them, we concluded that blood hyperviscosity in these animals is not their biological characteristics, Fig. 1. Effect of course intragastrical administration of LCE on whole blood viscosity at different shear rates. *p<0.05 in comparison with *Wistar rats, +Brattleboro rats (control).TABLE 1. Effect of Course Treatment with LCE on Macro- and Microrheological Parameters in Wistar and Brattleboro Rats (M±m)Parameter Wistar rats (n=8)Brattleboro ratscontrol (n=8) LCE (n=8)Plasma viscosity, mPa×sec 1.4±0.1 1.8±0.1* 1.6±0.1+Hematocrit, % 45±1 49±1* 49±1Fibrinogen concentration, g/liter 2.15±0.12 3.25±0.26* 2.33±0.14+Erythrocyte aggregation halftime, sec 10.1±0.5 5.9±0.2* 9.2±0.7+EIE 0.535±0.019 0.420±0.014* 0.500±0.011+O2 availability for tissues, arb. units 12.5±0.2 10.9±0.3* 11.6±0.3+Note. p<0.05 in comparison with *Wistar rats, +control.Bulletin of Experimental Biology and Medicine, Vol. 159, No. 5, September, 2015 METHODS691but a pathological condition, HBVS [6], caused by shifts in both macro- and microrheological parameters of the blood.Course treatment with LCE significantly im-proved both the macro- and microrheological param-eters of the blood in experimental rats: deformability of erythrocytes was improved in comparison with the control, EIE increased by 19% (Table 1), which can be due to antioxidant and antiradical activity of LCE [1]. Other substances with antiradical and antioxidant activity were also reported to increase erythrocyte de-formability [5,7,9]. Erythrocyte aggregation was also infl uenced by LCE (Table 1). In Brattleboro rats of the experimental group, a signifi cant (56%) increase in erythrocyte aggregation halftime was observed, which to a certain extent could be caused by reduced fi brino-gen content (28%). Plasma viscosity also decreased due to the decrease in fi brinogen content. Due to cumulative effect of LCE on the basic macro- and microrheological parameters, viscosity of the whole blood signifi cantly decreased by 4-15% in a wide range of shear rates (Fig. 1), which improved oxygen delivery to tissues by 6-12% in the studied range of shear rates (3-300 sec–1) without changes in hematocrit.Thus, under conditions of HBVS model in Brat-tleboro rats, course treatment with LCE (150 mg/kg intragastrically for 10 days) exhibited hemorheologi-cal activity and modulated macro- (plasma viscosity, fi brinogen concentration) and microrheological (eryth-rocyte aggregation and deformability) para meters. These fi ndings suggest that Brattleboro rats are a con-venient model of HBVS for the search and testing of substances with hemorheological activity.REFERENCES1. M. B. Plotnikov, L. N. Zibareva, A. A. Koltunov, et al., Rast. Resursy, 34, No. 1, 87-91 (1998).2. M. B. Plotnikov, O. I. Aliev, A. S. Vasil’ev, et al., Rast. Resursy, 35, No. 1, 103-107 (1999).3. M. B. Plotnikov, O. I. Aliev, M. Yu. Maslov, et al., Eksp. Klin. Farmakol., 63, No. 2, 54-56 (2000).4. M. B. Plotnikov, O. I. Aliev, A. S. Vasil’ev, et al., Bull. Exp. Biol. Med., 139, No. 1, 60-63 (2005).5. M. B. Plotnikov, N. A. Tyukavina, and T. M. Plotnikova, Di-quertin-Based Drugs [in Russian], Tomsk (2005).6. M. B. Plotnikov, A. S. Vasil’ev, O. I. Aliev, et al., Biomedit-sina, 1, No. 3, 57-62 (2011).7. V. Kucukatay, M. Bor-Kucukatay, G. Gundogdu, et al., Folia Biol. (Praha), 58, No. 4, 157-165 (2012).8. J. F. Stoltz and M. Donner, Schweiz. Med. Wochenschr., 43, Suppl., 41-49 (1991).9. H. Yapislar and E. Taskin, Med. Sci. Monit., 20, No. 1, 399-405 (2014).M. B. Plotnikov, A. S. Vasil’ev, et al.

Brattleboro rats as the model of blood hyperviscosity syndrome for testing substances with hemorheological activity

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Brattleboro rats as the model of blood hyperviscosity syndrome for testing substances with hemorheological activity

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