Hemostatic activity in vitro of lectin-containing substances of bryathypes on the example of some Antarctic and Belarus representatives

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Abstract

BACKGROUND: The modern market of medical devices in Belarus and Russia is represented by a wide range of hemostatic agents, of which the most popular are local hemostatics of plant origin possessing the significant technological potential for renewal and improvement. A promising reserve for this may be biologically active compounds of mosses, which are characterized by anti-inflammatory, antibacterial and antifungal effects. However, their hemostatic effect is almost not studied, which determines the relevance of this work.

AIM: The aim of this work is to study the effect of lectin-containing substances from mosses of three species collected in East Antarctica and Belarus on the parameters of human blood hemostasis in vitro.

MATERIALS AND METHODS: We studied mosses of the genera Bryum, Ceratodon, and Coscinodon, collected in the area of the Belarusian Antarctic station Gora Vechernyaya in East Antarctica and in Belarus. Lectin-containing substances of mosses were obtained by extracting shoots in 0.05 M tris-HCl buffer (pH 8.0), centrifugation, filtration. The assessment of the biological activity of lectin-containing substances in mosses was carried out by the agglutination reaction of rabbit erythrocytes, as well as the effect on human platelet aggregation and in the test for activated partial thromboplastin time.

RESULTS: It was established that lectin-containing substances of the studied moss species had agglutinating activity against erythrocytes in the range from 11708.28 (Belarusian samples) to 1333979.59 U/mg of protein (Antarctic samples) depending on the species and localization; initiated the aggregation of human platelets (25–80% of the effect of thrombin) regardless of blood group, Rh and gender of donors; influenced the plasma link of hemostasis, reducing activated partial thromboplastin time (by 15–18%).

CONCLUSIONS: It was found that some species of mosses of the genera Bryum, Ceratodon and Coscinodon of Antarctica and Belarus had an agglutinating and hemostatic effect on erythrocytes and platelets, with the greatest activity noted for Antarctic species. A hypothesis has been put forward that the observed phenomenon is due to the structural features of proteins, including lectins. It is assumed that lectins are possible inducers of erythrocyte agglutination and platelet aggregation in mosses. It is shown that the moss species Bryum pseudotriquetrum and Ceratodon purpureus have a certain resource potential in Belarus for their annual harvest. The results obtained expand the list of moss species with hemostatic activity, and can be used to develop new hemostatics of plant origin for local use from Belarusian plant materials.

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About the authors

Olga L. Kandelinskaya

Institute of Experimental Botany National Academy of Sciences of Belarus

Email: okandy@yandex.ru
ORCID iD: 0000-0002-0364-9548
SPIN-code: 2189-4733

Cand. Sci. (Biol.), Assistant Professor, Leading Researcher, Sector of Plant Protein Metabolism and Functions

Belarus, Minsk

Helena R. Grischenko

Institute of Experimental Botany National Academy of Sciences of Belarus

Email: helegreen@yandex.ru
ORCID iD: 0000-0002-2163-6759

Senior Research Associate of Sector of Plant Protein Metabolism and Functions

Belarus, Minsk

Daria V. Grigorieva

Belarusian State University

Email: dargr@tut.by
ORCID iD: 0000-0003-0210-5474
SPIN-code: 2479-7785

Cand. Sci. (Biol.), Assistant Professor of the Department of Biophysics, Physics Faculty

Belarus, Minsk

Irina V. Gorudko

Belarusian State University

Email: irinagorudko@gmail.com
ORCID iD: 0000-0002-4737-470X
SPIN-code: 8968-3125

Cand. Sci. (Biol.), Assistant Professor, Assistant Professor of the Department of Biophysics, Physics Faculty

Belarus, Minsk

Dmitriy V. Goreckiy

Institute of Experimental Botany National Academy of Sciences of Belarus

Email: goreckiydmitriy@yandex.by
ORCID iD: 0009-0000-8279-5094

Junior Research Associate of Sector of Plant Protein Metabolism and Functions

Belarus, Minsk

Eleonora V. Dashkevich

Republican Scientific and Practical Center for Transfusiology and Medical Biotechnologies of the Ministry of Health of the Republic of Belarus

Email: eleonoravdoc@gmail.com
ORCID iD: 0009-0000-9711-9371
SPIN-code: 1804-4804

Head of the Laboratory of Transfusiology

Belarus, Minsk

Janna V. Peshnyak

Republican Scientific and Practical Center for Transfusiology and Medical Biotechnologies of the Ministry of Health of the Republic of Belarus

Email: peshnyak@gmail.com
ORCID iD: 0009-0001-3709-3947
SPIN-code: 2496-2585

Leading Research Associate of the Laboratory of Transfusiology

Belarus, Minsk

Natallia A. Bukhvald

Republican Scientific and Practical Center for Transfusiology and Medical Biotechnologies of the Ministry of Health of the Republic of Belarus

Email: morskayaz300@mail.ru
ORCID iD: 0009-0003-4795-3271
SPIN-code: 8599-4504

Research Associate at the Laboratory of Transfusiology

Belarus, Minsk

Oleg M. Maslovsky

Institute of Experimental Botany National Academy of Sciences of Belarus

Email: oleg.maslovsky@tut.by
ORCID iD: 0009-0003-4976-5215
SPIN-code: 7756-0934

Cand. Sci. (Biol.), Head of Sector of Plant Cadastre

Belarus, Minsk

Irina P. Sysoi

Institute of Experimental Botany National Academy of Sciences of Belarus

Email: mastibrotskaya@gmail.com
ORCID iD: 0009-0001-7777-9433
SPIN-code: 1719-9379

Cand. Sci. (Biol.), Senior Research Associate of Sector of Plant Cadastre

Belarus, Minsk

Yury G. Hihiniak

Scientific and Practical Center for Bioresources National Academy of Sciences of Belarus

Email: antarctida_2010@mail.ru
ORCID iD: 0000-0001-8376-3991

Cand. Sci. (Biol.), Assistant Professor, Leading Research Associate of the Sector for Monitoring and Cadastre of the Wildlife

Belarus, Minsk

Egor V. Korzun

Scientific and Practical Center for Bioresources National Academy of Sciences of Belarus

Email: natrix109@gmail.com
ORCID iD: 0009-0002-9318-9625
SPIN-code: 9108-1529

Senior Research Associate of Sector of monitoring and wildlife cadastre

Belarus, Minsk

Valeria A. Kostevich

Institute of Experimental Medicine; Academician Yu.M. Lopukhin Federal Scientific and Clinical Center for Physical and Chemical Medicine of the Federal Biomedical Agency

Email: hfa-2005@yandex.ru
ORCID iD: 0000-0002-1405-1322
SPIN-code: 2726-2921

Cand. Sci. (Biol.), Senior Research Associate of the Department of Molecular Genetics; Research Associate fellow of Department of Biophysics

Russian Federation, Saint Petersburg; Moscow

Mikhail P. Andreev

V.L. Komarov Botanical Institute of the Russian Academy of Sciences

Email: andreev@gmail.com
ORCID iD: 0000-0002-5688-0751
SPIN-code: 6075-6128

Head of the Laboratory of Lichenology and Bryology

Russian Federation, Saint Petersburg

Lyubov E. Kurbatova

V.L. Komarov Botanical Institute of the Russian Academy of Sciences

Author for correspondence.
Email: kurbatovae@binran.ru
ORCID iD: 0000-0003-4695-5331
SPIN-code: 2032-8596

Senior Research Associate of the Laboratory of Lichenology and Bryology

Russian Federation, Saint Petersburg

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2. Fig. 1. Moss Bryum pseudotriquetrum: a — distribution map (https://www.gbif.org/species/2676867); b — curtain; с — shoots (photos courtesy of L.E. Kurbatova)

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3. Fig. 2. Moss Ceratodon purpureus: a — distribution map (https://www.gbif.org/species/5281381); b — network on low rocky outcrops; c — shoots (photographs by L.E. Kurbatova)

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4. Fig. 3. Moss Coscinodon lawianus: a — distribution map (https://www.gbif.org/species/8124837); b — sods; c — shoots (photographs by L.E. Kurbatova)

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5. Fig. 4. Typical kinetic curves of platelet aggregation with the addition of Antarctic moss lectin-containing substances (a–c) at various concentrations and thrombin (d). Platelets (2.5 · 10^8 cells/ml) in PBS containing 1 mM CaCl2, 0.5 mM MgCl2. Sample numbers: No. 1 — B. pseudotriquetrum, No. 2 — C. purpureus, No. 3 — C. lawianus. Concentrations used in figure a: 1 — 0.11, 2 — 0.23, 3 — 0.57, 4 — 1.14, 5 — 2.27 µg/ml; b: 1 — 0.18, 2 — 0.45, 3 — 0.9, 4 — 1.8 µg/ml; с: 1 — 0.12, 2 — 0.3, 3 — 0.6, 4 — 1.2 µg/ml; d — 0.5 µg/ml

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6. Fig. 5. Dependence of the parameters of platelet aggregation (a — degree, b — rate) initiated by the addition of Antarctic moss lectin-containing substances, on the concentration of these compounds. Platelets (2.5 · 10^8 cells/ml) in PBS containing 1 mM CaCl2, 0.5 mM MgCl2. Sample numbers: No. 1 — B. pseudotriquetrum, No. 2 — C. purpureus, No. 3 — C. lawianus

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7. Fig. 6. Typical kinetic curves of platelet aggregation with the addition of Belarus moss lectin-containing substances at various concentrations. Platelets (2.5 · 10^8 cells/ml) in PBS containing 1 mM CaCl2, 0.5 mM MgCl2. Sample numbers: No. 4 — B. pseudotriquetrum, No. 5 — C. purpureus. Concentrations used in figure a: 1 — 3.4, 2 — 8.5, 3 — 17, 4 — 34 µg/ml; b: 1 — 1.4, 2 — 3.6, 3 — 7.1, 4 — 14.3 µg/ml

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8. Fig. 7. Dependence of the parameters of platelet aggregation (a — degree, b — rate) initiated by the addition of Belarus moss lectin-containing substances, on the concentration of these compounds. Platelets (2.5 · 10^8 cells/ml) in PBS containing 1 mM CaCl2, 0.5 mM MgCl2. Sample numbers: No. 4 — B. pseudotriquetrum, No. 5 — C. purpureus

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9. Fig. 8. Distribution of exploitable stocks of B. pseudotriquetrum raw materials on the territory of the Republic of Belarus by administrative regions

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