Study of the biostimulating effect of a decoction of Michurin's chokerenia leaves in vitro using the Parameсium caudatum cilia test system

Introduction. Aronia × mitschurinii A. K. Skvortsov & Maitul. is a cultigenic species bred in the late 19th century as a result of breeding work by I. V. Michurin by crossing Aronia melanocarpa (Michx.) Elliott with distantly related species of the rowan genus (Sorbus). The fruits of this plant have long been used in folk and traditional medicine, being, first of all, a source of anthocyanin compounds. The leaves are currently only a by-product in the preparation of fruits, however, they can be used as a promising source of such groups of biologically active substances as flavonoids, tannins, leucoanthocyanins, which makes it possible to use this raw material to obtain herbal medicinal products (HMP). Therefore, primary in vitro screening studies to assess the toxicity and biostimulating effect of extracts from the leaves of Aronia michurini should be considered relevant to assess the prospects and determine the directions of subsequent preclinical trials. The aim of the study was to investigate the biostimulating and membrane-protective effect of Michurin's chokeberry leaf extracts (using a decoction as an example) in vitro using the Parameсium caudatum ciliate test system in the «functional load» test.

Materials and methods. Michurin's chokeberry leaves were harvested for the study at different periods of leaf blade development (May, June, August and September 2023) from plants cultivated on the territory of the I. V. Michurin Federal Scientific Center (Michurinsk, Tambov Region). For the study, aqueous extracts were prepared from the studied raw materials as a decoction in accordance with general pharmacopoeial monograph.1.4.1.0018.15 «Infusions and decoctions» for raw materials containing tannins. To study the biostimulating effect of the decoction of Michurin's chokeberry leaves, an in vitro test system of Parameсium caudatum ciliates in the stationary growth phase was used. The biostimulating and membrane-protective effect of the studied decoction was assessed in the «functional load» test under the damaging effect of a hypertonic sodium chloride solution.

Results and discussion. Using the test system of the Parameсium caudatum ciliates in the «functional load» test, it was established that a decoction of Michurin’s chokeberry leaves in vitro in a 1 : 10 dilution at all times of the medicinal plant raw material preparation reduces the survival time of ciliates under the damaging effects of a hypertonic sodium chloride solution during an incubation of 24 hours by no less than 43.5 % compared to the control sample, which probably indicates an increase in the permeability of biological membranes under the influence of tannins and sterols of the leaves and is consistent with their known astringent and antibacterial action. A decoction of Michurin's aronia leaves (when harvesting medicinal plant raw materials in June) in vitro in dilutions of 1 : 1000 and 1 : 10 000 close to the ranges of therapeutic doses, exhibits the greatest biostimulating and membrane-protective effect, which is confirmed by a significant reliable increase in the survival time of ciliates under the damaging effects of hypertonic sodium chloride solution, by a maximum of 39.3 % compared to the control sample during 24 h incubation, which is probably associated with the content of flavonoids and leucoanthocyanins and is consistent with their known capillary-protective and antioxidant effects.

Conclusion. A decoction of Michurin's chokeberry leaves (diluted 1 : 1000 when harvesting medicinal plant raw materials in June) significantly and reliably increases the survival time of the Parametium caudatum ciliate test system under the damaging effects of sodium chloride solution in the «functional load» test, by a maximum of 39.3 % compared to the control sample during 24-hour incubation, which indicates the presence of a biostimulating and membrane-protective effect.

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