The Mechanisms of Lipid Vesicle Fusion Inhibition by Extracts of Chaga and Buckthorn Leaves

Abstract

The ability of extracts of grapefruit seeds (ESG), sea buckthorn leaves (ESBL), and chaga (EC) to inhibit membrane fusion was evaluated. It was found that ESBL and EC inhibited Ca²⁺-mediated fusion of phosphatidylglycerol-enriched lipid vesicles; the inhibition indexes were about 90 and 100%, respectively. ESG did not inhibit the fusion of negatively charged liposomes induced by calcium. In addition to calcium-mediated liposome fusion, EC inhibited the fusion of vesicles from a mixture of phosphatidylcholine and cholesterol under the action of polyethylene glycol with a molecular weight of 8000 Da (the inhibition index was 80%). The other two extracts had no effect on polymer-induced fusion of uncharged membranes. The effect of some major components of the tested extracts on the fusion of vesicles was evaluated. It has been shown that flavonols, quercetin and myricetin, which are major components of ESBL, inhibited the fusion of negatively charged membranes under the action of calcium (the inhibition indexes were about 85 and 60%, respectively). Another flavonol of ESBL, the glycoside of quercetin rutin, did not have such an effect. The data obtained made it possible to relate the ESBL suppression of calcium-induced fusion of lipid vesicles with the presence of quercetin and myricetin in its composition. These flavonols had virtually no effect on polyethylene glycol-induced vesicle fusion, which is consistent with the absence of ESBL action on liposome fusion under the action of polymer. The ability of quercetin and myricetin to reduce the melting temperature of phosphatidylglycerol with saturated hydrocarbon chains and to increase the half-width of the peak corresponding to melting has been demonstrated. The observed correlation between the parameters characterizing the thermotropic behavior of the lipid in the presence of quercetin and myricetin and the index of inhibition of calcium-mediated liposome fusion by these compounds may indicate a relationship between the ability of flavonols to influence the packaging of membrane lipids and inhibit vesicle fusion. Pentacyclic triterpenoids, betulin and lupeol, which are part of EC, did not inhibit the fusion of vesicles under the action of both calcium and polyethylene glycol, and their presence in EC cannot be responsible for the antifusogenic activity of EC.

Keywords: flavonols; liposomes; membrane fusion inhibitors; natural extracts; saponins.