Page 130 - Book of Abstracts

5 International Scientific Online Conference DOI: https://doi.org/10.15414/2021.9788055224015

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IN VITRO ANTIOXIDANT RESPONSE OF THE EQUINE BLOOD TREATED BY EXTRACT
OBTAINED FROM FICUS DRUPACEA THUNB. LEAVES (MORACEAE)
Halyna Tkachenko , Lyudmyla Buyun , Natalia Kurhaluk , Vitaliy Honcharenko ,
1
2
1
3
Andriy Prokopiv
3,4
1 Institute of Biology and Earth Sciences, Pomeranian University in Słupsk, Poland;
E-mail.: halyna.tkachenko@apsl.edu.pl
2 M.M. Gryshko National Botanic Garden, National Academy of Science of Ukraine, Kyiv, Ukraine
3 Ivan Franko National University of Lviv, Ukraine
4 Botanic Garden of Ivan Franko National University of Lviv, Ukraine
The objective of this report is to highlight the antioxidant properties of the leaf extract
obtained from Ficus drupacea Thunb. The degree of amelioration of oxidative damage was
determined using the equine plasma and erythrocytes model through in vitro bioassay by
comparing the effects of leaf extract on the superoxide dismutase, catalase, and glutathione
peroxidase activities.
The leaves of F. drupacea were collected in M.M. Gryshko National Botanical Garden (Kyiv,
Ukraine). Freshly collected leaves were washed, weighed, crushed, and homogenized in 0.1M
phosphate buffer (pH 7.4) (in proportion 1:19, w/w) at room temperature. The extracts were
then filtered and used for analysis. The pellet of blood was resuspended in 4 mM phosphate
buffer (pH 7.4). A volume of 0.1 ml of the plant extract was added to 1.9 ml of equine
erythrocytes or 1.9 ml of plasma. For positive control (4 mM phosphate buffer) was used. After
incubating the mixture for 60 min at 37 °C with continuous stirring, it was centrifuged at 3,000
rpm for 5 min. Erythrocytes aliquots were used in the study. Superoxide dismutase (SOD, E.C.
1.15.1.1) activity was assessed by its ability to dismutate superoxide produced during quercetin
auto-oxidation in an alkaline medium (pH 10.0) by Kostiuk et al. (1990). Catalase (CAT, E.C.
1.11.1.6) activity was determined by the method of Koroliuk et al. (1988). Glutathione
peroxidase (GPx, EC 1.11.1.9) activity was determined according to the method of Moin (1986).
The ceruloplasmin (CP, EC 1.16.3.1) level in the plasma was measured spectrophotometrically,
as described by Ravin (1961). The TAC level in the samples was estimated by measuring the 2-
thiobarbituric acid reactive substances (TBARS) level after the Tween 80 oxidation according
to Galaktionova et al. (1998). All statistical calculation was performed on separate data with
STATISTICA 8.0 software (StatSoft, Krakow, Poland).
In our study, the aqueous leaf extract of F. drupacea has proven effective to increase the
SOD, catalase, and GPx activity (by 41.6 %, 32.4 %, and 61.5 %, p<0.05). The increase of the
SOD, catalase, and GPx activity was induced by TAC enhancement in the erythrocytes and
plasma samples (p>0.05). SOD activity was non-significantly increased by 4 % (p>0.05). On the
other hand, F. drupacea leaf extract caused a statistically significant decrease in ceruloplasmin
level by 58.7 % (p<0.05). The results of this research indicated that crude extract obtained from
F. drupacea leaves has an effective antioxidant effect after treatment of a suspension of equine
erythrocytes. The protective effect of F. drupacea extract is evident by amelioration in activities
of antioxidant enzymes (SOD, catalase, and GPx). The pronounced effect of F. drupacea leaf
extract, probably, could be attributed to its secondary metabolites content, e.g. polyphenols and
flavonoids contents. It is believed, that there is a substantial discrepancy between findings
obtained in vitro and in vivo experiments. Therefore, further in vivo investigation is necessary
to reveal the exact cellular mechanisms of the effect of F. drupacea extract on the erythrocyte
membrane function.
Keywords: Ficus drupacea, leaves, extract, antioxidant enzymes, total antioxidant capacity.

Acknowledgments
We are grateful to The Polish National Commission for UNESCO for supporting our study.

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Spiritual Human Development
November 3 2021
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