For investigation of biologically active substance (BAS) effects the consistent number of experimental objects was used: model - multilamellar phospholipid liposomes (MPL), formed from synthetic individual phospholipid dimyristoylphosphatidylcholine (DMPC), and from mix of natural phospholipids of egg lecithin; organelles - fragmented sarcoplasmic reticulum (FSR) and erythrocyte’s ghost (plasma membrane with cytoskeleton); cells-insulated erythrocytes and cells of Ehrlich ascetic carcinoma (EAC). The biologically active substances, which were used: at first - the exogenous substances: melafen, caffeine and its analogs and antagonists, phenosan and its derivatives; at second - endogenic substances: free fatty acids (FFA), serum albumin, ATP and inorganic salts, containing magnesium and calcium. The registration of experimental objects reactions to the actions of BAS was held by the methods: DSC, small-angle x-ray light scattering (SAXS), primary light scattering at a right angle in the visible range, potentiometric methods. The concentration regions of BAS have been determined by DSC under the application of phenoksan (10-5 M) and IHFANs (10-6 M). These regions were without the destruction of microdomain organization at bilayers of MPL, formed from DMPC. Melafen over a wide range of concentrations didn’t disrupt the structure of such MPL, but caused the changes of thermodynamic parameters. IHFAN-10 and phenoksan changed the parameters of thermo-induced denaturation of protein microdomains of erythrocyte ghosts, melafen hadn’t effect. SAXS didn’t show of melafen influence at wide range of concentrations on the bilayers thickness and packing at natural MPL. The concentration-dependent actions of Mg2+ to the activities of Са2+-ATPase and Ca2+-channel have been showed by potentiometric method, also with the presence of FFA in FSR membranes. FFA and Mg2+ influenced to the effect of Ca2+-channel activator - caffeine. The extraction of FFA from FSR by the treatment with serum albumin reduced the effect of caffeine. Primary light diffusion showed the inhibition of purine-depended Ca2+-signalling at EAC cells by the melafen and phenoksan. It has been concluded that the approach based on the using of these experimental methods and objects could be applied to the substances test with the aim of revealing of their biological activity.
phospholipids, multilamellar liposomes, biology active substances, erythrocyte’s ghost, erythrocyte, DSC, small-angle X-ray scattering
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