The influence of metabolites isolated from Empetrum nigrum L. on the hemostasis system in vitro

DOI: https://doi.org/10.29296/25419218-2023-08-04
Issue: 
8
Year: 
2023

V.G. Luzhanin(1), A.V. Samorodov(2), A.K. Whaley(3), A.O. Whaley(3, 4), G.P. Yakovlev(3), I.A. Samylina(5)
1-Perm State Pharmaceutical Academy, st. Ekaterininskaya, 110, Perm, 614990, Russian Federation;
2-Bashkir State Medical University of the Ministry of Health of the Federation, st. Lenina, 3, Ufa, 450008, Russian Federation;
3-Saint Petersburg State Chemical Pharmaceutical University, st. Prof. Popova, 14A, St. Petersburg, 197376, Russian Federation;
4-Laboratory of Cellular Mechanisms of Blood Homeostasis, Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences, Toreza Ave., 44, St. Petersburg, 194223, Russian Federation;
5-A.P. Nelyubin Institute of Pharmacy of the I.M. Sechenov First Moscow State Medical University of the Ministry of Health of Russia (Sechenov University), st. Trubetskaya, 8, building 2, Moscow, 119991, Russian Federation

Introduction. Empetrum nigrum L. produces a wide variety of secondary metabolites of polyphenolic nature. According to the literature, E. nigrum exhibits antitumor, antidiabetic, and hypolipidemic activity, due to which it can be used for the treatment and prevention of relevant diseases. The development of thrombosis is a significant complication that occurs in metabolic disorders and cancer, for the prevention and treatment of which it is necessary to search for new biologically active substances. Purpose of the study. The purpose of this work is to screen individual components 1–19 isolated from E. nigrum shoots in relation to the hemostatic system. Material and methods. In vitro experiments were performed on the blood of healthy male donors aged 18–24 years. The study of the effect on platelet aggregation was carried out using the Born method on an AT-02 aggregometer (NPF Medtech, Russia). Determination of anticoagulation activity was carried out by generally accepted clotting tests on a turbidimetric hemocoagulometer Solar CGL 2110 (ZAO SOLAR, Russia). As a marker of platelet activation, the expression of P-selectin on the surface of platelets was measured using a cytofluorometric method using a NovoCyte device (Agilent, USA). Results. Compounds 1, 4, 5, 6, 11, 13, 15 and 18 showed antiaggregation activity at the level of acetylsalicylic acid in vitro, while compounds 3, 8 and 12 showed antiaggregation activity exceeding the values of acetylsalicylic acid. It was also found that compounds 9 and 14 did not affect the level of CD62 expression. Conclusion. As a result of screening individual compounds 1-19 isolated from E. nigrum shoots, their antiaggregation and anticoagulation properties were established.
Keywords: 
Empetrum nigrum L.
black crowberry
polyphenols
homeostasis.
References: 
  1. Stevens P.F., Luteyn J., Oliver E.G.H., Bell T.L., Brown E.A., Crowden R.K., George A.S., Jordan G.J., Ladd P., Lemson K., McLean C.B., Menadue Y., Pate J.S., Stace H.M., Weiller C.M. "Ericaceae". In Kubitzki, K. (ed.). Flowering Plants. Dicotyledons: Celastrales, Oxalidales, Rosales, Cornales, Ericales. The families and genera of vascular plants. 6. Springer. 2004; 145–94.
  2. Maatta-Riihinen K.R., Kamal-Eldin A., Mattila P.H., Gonzalez-Paramas A.M., Torronen A.R. Distribution and contents of phenolic compounds in eighteen Scandinavian berry species. Journal of agricultural and food chemistry. 2004; 52 (14): 4477–86. DOI: 10.1021/jf049595y.
  3. Ponkratova A.O., Whaley A.K., Balabas O.A., Smirnov S.N., Proksch P., Luzhanin V.G. A new bibenzyl and 9,10-dihydrophenanthrene derivative from aerial parts of crowberry (Empetrum nigrum L.). Phytochemistry Letters. 2021; 42: 15–7. DOI: 10.1021/jf049595y.
  4. Ponkratova A.O., Whaley A.K., Orlova A.A., Smirnov S.N., Serebryakov E.B., Proksch P., Luzhanin V.G. A new dimethoxy dihydrochalcone isolated from the shoots of Empetrum nigrum L. Natural Products Research. 2022; 36 (20): 5142–7.
  5. Li H., Jean S., Webster D., Robichaud G.A., Calhoun L.A., Johnson J.A., Gray C.A. Dibenz[b,f]oxepin and Antimycobacterial Chalcone Constituents of Empetrum nigrum. Journal of natural products. 2015; 78 (11): 2837–40. DOI: 10.1021/acs.jnatprod.5b00627.
  6. Krasnov E.A., Ermilova E.V., Kadyrova T.V., Druganov A.G., Semenov A.A., Tolstikov G.A. Phenolic components of Empetrum nigrum extract and the crystal structure of one of them. Chemistry of natural compounds. 2000; 36 (5): 493–6. DOI: 10.1023/A:1002887406817.
  7. Понкратова А.О., Уйэли А.К., Жохова Е.В., Безверхняя Е.А., Лужанин В.Г. Сравнительный фитохимический анализ образцов надземной части Empetrum nigrum L., собранных в различных регионах РФ, как перспективного источника фармакологически активных вторичных метаболитов. Разработка и регистрация лекарственных средств. 2021; 10 (4): 138–45. DOI: 10.33380/2305-2066-2021-10-4(1)-138-145. [Ponkratova A.O., Whaley A.K., Zhokhova E.V., Bezverkhniaia E.A., Luzhanin V.G. Comparative phytochemical analysis of the aerial parts of Empetrum nigrum L. samples, collected in various regions of the Russian Federation. Drug development & registration. 2021; 10 (4): 138–45. DOI: 10.33380/2305-2066-2021-10-4(1)-138-145 (In Russ.)]
  8. Понкратова А.О., Уэйли А.К., Лужанин В.Г., Жохова Е.В. Использование метода высокоэффективной тонкослойной хроматографии для обнаружения фармакологически активных вторичных метаболитов в водянике черной Empetrum nigrum L. Разработка и регистрация лекарственных средств. 2021; 10 (4): 129–37. DOI: 10.33380/2305-2066-2021-10-4(1)-129-137. [Ponkratova A.O., Whaley A.K., Luzhanin V.G., Zhokhova E.V. Using high performance thin layer chromatography for the detection of pharmacologically active secondary metabolites in Empetrum nigrum L. Drug development & registration. 2021; 10 (4): 129–37. DOI: 10.33380/2305-2066-2021-10-4(1)-129-137 (In Russ.)]
  9. Whaley A., Minakov D., Orlova A., Ponkratova A., Fock E., Rukoyatkina N., Gambaryan S., Luzhanin V. Analysis of Empetrum nigrum L. lipophilic secondary metabolites, their metabolomic profiles and antioxidant activity. Journal of Essential Oil Research. 2023; 35: 1-14. DOI:10.1080/10412905.2023.2169377
  10. Oka S., Kuniba R., Tsuboi N., Tsuchida S., Ushida K., Tomoshige S., Kuramochi K. Isolation, synthesis, and biological activities of a bibenzyl from Empetrum nigrum var. japonicum. Bioscience, biotechnology, and biochemistry. 2020;8 4 (1): 31-36. DOI:10.1080/09168451.2019.1662279
  11. Kovacs A., Vasas A., Hohmann J. Natural phenanthrenes and their biological activity. Phytochemistry. 2008; 69 (5): 1084-1110. DOI:10.1016/j.phytochem.2007.12.005
  12. Singh P., Anand A., Kumar V. Recent developments in biological activities of chalcones: a mini review. European journal of medicinal chemistry. 2014; 85: 758-777. DOI:10.1016/j.ejmech.2014.08.033
  13. Matsuura H., Saxena G., Farmer S.W., Hancock R.E.W., Towers G.H.N. Antibacterial and antifungal compounds from Empetrum nigrum. Planta Medica. 1995; 61: 580. DOI: 10.1055/s-2006-959382
  14. Moerman D.E. Native American Medicinal Plants: An Ethnobotanical Dictionary. Timber Press: Portland, OR. 2009; 799.
  15. Huttunen S., Toivanen M., Arkko S., Ruponen M., Tikkanen Kaukanen C. Inhibition activity of wild berry juice fractions against Streptococcus pneumoniae binding to human bronchial cells. Phytotherapy Research. 2011; 25 (1): 122–127.
  16. Kim K.C., Lee I.K., Kang K.A., Kim B.J., Kim D., Moon J.Y., Hyun J.W. Empetrum nigrum var. japonicum extract suppresses γ-ray radiation-induced cell damage via inhibition of oxidative stress. The American journal of Chinese medicine. 2011; 39 (01): 161–170.
  17. Nohynek L.J., Alakomi H.L., Kähkönen M.P., Heinonen M., Helander I.M., Oksman-Caldentey K.M., Puupponen-Pimiä R.H. Berry phenolics: antimicrobial properties and mechanisms of action against severe human pathogens. Nutrition and Cancer. 2006; 54 (1): 18–32.
  18. Hyun T.K., Kim H.C., Ko Y.J., Kim J.S. Antioxidant, α-glucosidase inhibitory and anti-inflammatory effects of aerial parts extract from Korean crowberry (Empetrum nigrum var. japonicum). Saudi Journal of Biological Sciences. 2016; 23 (2): 181–188.
  19. Kim K.C., Kim D., Kim S.C., Jung E., Park D., Hyun J.W. Empetrum nigrum var. japonicum extract suppresses ultraviolet B-induced cell damage via absorption of radiation and inhibition of oxidative stress. Evid. Based Complement. Altern. Med. 2013: 983609.
  20. Törrönen R., McDougall G.J., Dobson G., Stewart D., Hellström J., Mattila P., Pihlava J.M., Koskela A., Karjalainen R. Fortification of blackcurrant juice with crowberry: Impact on polyphenol composition, urinary phenolic metabolites, and postprandial glycemic response in healthy subjects. Journal of Functional Foods. 2012; 4: 746–756. DOI: 10.1016/j.jff.2012.05.001
  21. Kellogg J., Wang J., Flint C., Ribnicky D., Kuhn P., De Mejia E.G., Raskin I., Lila M.A. Alaskan wild berry resources and human health under the cloud of climate change. J. of Agricultural and Food Chemistry. 2010; 58: 3884–900. DOI: 10.1021/jf902693r
  22. Jurikova T, Mlcek J, Skrovankova S, Balla S, Sochor J, Baron M, Sumczynski D. Black crowberry (Empetrum nigrum L.) flavonoids and their health promoting activity. Molecules. 20167; 21 (12): 1685. DOI:10.3390/molecules21121685
  23. Kodiatte T.A., Manikyam U.K., Rao S.B., Jagadish T.M., Reddy M., Lingaiah H.K.M., Lakshmaiah V. Mean platelet volume in type 2 diabetes mellitus. Journal of Laboratory Physicians. 2012; 4 (1): 5-9. DOI:10.4103/0974-2727.98662
  24. Barbui T, De Stefano V, Falanga A, Finazzi G, Martinelli I, Rodeghiero F, Vannucchi A.M, Barosi G. Addressing and proposing solutions for unmet clinical needs in the management of myeloproliferative neoplasm-associated thrombosis: A consensus-based position paper. Blood Cancer Journal. 2019; 9 (8): 61. DOI:10.1038/s41408-019-0225-5
  25. Понкратова А.О., Уйэли А.К., Жохова Е.В., Безверхняя Е.А., Лужанин В.Г. Сравнительный фитохимический анализ образцов надземной части Empetrum nigrum L., собранных в различных регионах РФ, как перспективного источника фармакологически активных вторичных метаболитов. Разработка и регистрация лекарственных средств. 2021; 10 (4): 138–45. DOI: 10.33380/2305-2066-2021-10-2-80-86. [Ponkratova A.O., Whaley A.K., Orlova A.A., Smirnov S.N., Serebryakov E.B., Luzhanin V.G. Isolation and structure elucidation of three dimeric A-type proanthocyanidins from Empetrum nigrum L. Drug development & registration. 2021; 10 (2): 80–6. DOI: 10.33380/2305-2066-2021-10-2-80-86 (In Russ.)]
  26. Миронов А.Н. и др. Руководство по проведению доклинических исследований лекарственных средств/ М.: Гриф и К., 2012; 944. [Mironov A.N. et al. Rukovodstvo po provedeniyu doklinicheskih issledovanij lekarstvennyh sredstv. CHast' pervaya. M.: Grif i K., 2012; 944 (In Russ.)]
  27. Born G.G.V. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature.1962; 194: 927–9.