Turkina V. A., Yavorska N. Y., Lapovets N. Y., Vorobets N. M, Vishchur O. I.

BIOLOGY

Scentific article

Abstract. Plants are characterized by organ-specific composition and content of polyphenolic compounds, and they are largely responsible for changes in the immune homeostasis of mammals. At the same time, shoots of Vaccinium corymbosum as a by-product of its cultivation have not been studied from the standpoint of their extract possible immunotropicity. The aim of this study was to conduct a primary immunological screening of extracts of shoots of Vaccinium corymbosum L. in vivo. Shoots of blueberry V. corymbosum of the Elliott variety grown on the experimental plot were used for research. Aqueous-ethanol extract was prepared using 60% aqueous ethanol as extractant by maceration (1:10 / weight: volume / g: ml); aqueous – the extract was prepared by suspending in distilled water under reflux on a low-boiling water bath. The experimental work was performed on 24 outbred male albino guinea pigs, which were kept in vivarium. By the method of «blind ranking» animals were divided into three groups: the 1st group – control, the 2nd group – animals sensitized with water-ethanol extract, the 3rd group – sensitized with water extract. Extracts in a volume of 0.02 ml were injected once intradermally into the outer surface of the ear of experimental animals. After 10 days, additional epicutaneous applications were performed on pre-depilated areas of the lateral surface of the animal’s body 5 times a week for 4 weeks. Blood was taken by cardiac puncture to quantify immune homeostasis in anesthetized experimental animals. Changes in peripheral blood parameters (absolute number of leukocytes and leukocyte formula) were determined and hematological indices were calculated. Phenotyping of peripheral blood lymphocytes was performed by indirect immunofluorescence determination using monoclonal antibodies. Population and subpopulations of lymphocytes were counted using a fluorescence microscope with a phase contrast attachment. Immunoglobulin E in blood serum was determined by immunoassay analysis. Circulating immune complexes were investigated by the method of precipitation spectrophotometrically. Statistical processing of the results was carried out using the Microsoft Excel software package. In the group of animals affected by water-ethanol extract there was a tendency to decrease the total number of leukocytes, at the same time the effect of aqueous extract caused a tendency to increase this index, but in comparison with the control group there were no significant differences. The redistribution of leukocyte cell pools in both experimental groups was revealed in comparison with the control. It was found that under the influence of water-ethanol extract of V. corymbosum shoots in animals, the absolute number of lymphocytes decreases. The detected increase in the number of CD4 cells indicates an increase in the reactivity of lymphocytes and the dominance of T-helpers, an increase in the number of CD8 indicates the activation of the killer link of immunity. In animals of the 2nd and 3rd experimental groups, an increase was found in comparison with the control of the level of Ig E immunoglobulins. The obtained results indicate the development of type 1 hypersensitivity reactions in animals of the experimental groups.

Vaccinium corymbosum, immunity, guineapig.

1. Jantan I, Ahmad W, Bukhari SN. Plant-derived immunomodulators: An insight on their preclinical evaluation and clinical trials. Frontiers in Plant Science. 2018;9:1178. DOI: 10.3389/fpls.2018.01178
2. Mlcek J, Jurikova T, Skrovankova S, Sochor J. Quercetin and Its Anti-Allergic Immune Response. Molecules. 2016;21(5):623. DOI: 10.3390/ molecules21050623
3. Wang LJ, Wu J, Wang HX, Li SS, Zheng XC, Du H, et al. Composition of phenolic compounds and antioxidant activity in the leaves of blueberry cultivars. Journal of Functional Foods. 2015;16:295–304.
4. Sun Y, Li M, Mitra S, Hafiz MR, Debnath B, Lu X, et al. Comparative Phytochemical Profiles and Antioxidant Enzyme Activity Analyses of the Southern Highbush Blueberry (Vaccinium corymbosum) at Different Developmental Stages. Molecules. 2018;23(9):2209. DOI: 10.3390/ molecules23092209
5. Giovanelli G, Buratti S. Comparison of polyphenolic composition and antioxidant activity of wild Italian blueberries and some cultivated varieties. Food Chemistry. 2009;112(4):903–8. DOI: 10.1016/j.foodchem.2008.06.066
6. Ovalle-Marin A, Reyes-Farias M, Vasquez K, Parra-Ruiz C, Quitral V, Jimenez P, et al. Maqui, Calafate, and Blueberry fruits extracts treatments suppress the pathogenic interaction amongst human adipocytes and macrophages. Journal of Berry Research. 2020;10(3):531– 45. DOI: 10.3233/ jbr-200576
7. Wei L, Chun-peng W. Research progresses on blueberry of phytochemical constituents and activities in recently ten years. The Food Industry. 2015;36(10):233–7.
8. Riihinen K, Jaakola L, Kärenlampi S, Hohtola A. Organ-specific distribution of phenolic compounds in bilberry (Vaccinium myrtillus) and ‘northblue’ blueberry (Vaccinium corymbosum x V. angustifolium). Food Chemistry. 2008;110(1):156–60. DOI: 10.1016/j. foodchem.2008.01.057
9. Yavors’ka NY, Vorobets’ NM. Vmist polifenoliv ta flavonoyidiv u pahonakh lokhyny vysokorosloyi protyahom vehetatsiynoho periodu. Visnyk problem biolohiyi i medytsyny. 2020;3(157):70-75. DOI: 10.29254 /2077-4214-2020-3-157-70-75. [in Ukrainian]
10. Shi D, Xu M, Ren M, Pan E, Luo C, Zhang W, et al. Immunomodulatory Effect of Flavonoids of Blueberry (Vaccinium corymbosum L.) Leaves via the NF-κB Signal Pathway in LPS-Stimulated RAW 264.7 Cells. Journal of Immunology Research. 2017;2017:1–7. DOI: 10.1155/2017/5476903
11. Treaty Office. Full list [Internet]. 2021 [cited 2021 Feb 26]. Available from: https://www.coe.int/en/web/conventions/full-list/-/conventions/ treaty/123
12. Lapovets’ LYe, Lutsyk BD, Lebed’ HB, Akimova VM, Zubchenko SO. Posibnyk z laboratornoyi imunolohiyi. L’viv; 2014. 292 s. [in Ukrainian].
13. Grinevich YUA, Alferov AI. Opredeleniye immunnykh kompleksov. Laboratornoye delo. 1981;8:493-96. [in Russian].
14. Cerutti A, Cols M, Puga I. Marginalzone B cells: virtuesof innate-like antibody-producing lymphocytes. Nat Reviews Immunology. 2013;13:118–132. DOI: https://doi.org/10.1038/nri3383

«Bulletin of problems biology and medicine» Issue 1 (159), 2021 year, 143-147 pages, index UDK 611-092.4/9:612.017.2:615.322

10.29254/2077-4214-2021-1-159-143-147