Methodical approaches for increasing of experimental objects answers under biological active substances actions in vitro

Alekseeva O.M., Krementsova A.V., Golochshapov A.N.

N.M. Emanuel Institute of Biochemical Physics of Russian Academy of Sciences, Russia, 119334, Moscow, Kosygin St. 4, +74959397409,

It is known that physiologically the adequate responses of biological structures of animal origin by biologically active substance (BAS) in numerous cases not are available for registration in connection with little the parameter change of experimental object. This is why in operating in vitro with model biologic objects was is selected condition for intensification of answer of experimental biologic object to get logged result. As BAS we used the synthetic material melafen (the melamine salt of the bisphophinic acid) - the plant growth regulator, and the nature substance methylxanthine - caffeine. As the model objects used lipid and protein-lipids membrane, organelle and cell. Study on the effects of melafen on structural property of biomembrane was held by means of approaches, intended to strengthen sensibility of experimental biologic objects to action. The melafen influences on membrane structure were tested by means of differential scanning microcalorymetry (DSK). Lipid-lipid interactions under the impact of melafen logged when melting of multilamellar liposomes from dimyristoilphosphatidylcholine (DMPC). The speed variation of heat supplied to models allowed to strengthen and to educe the melafen effects on restructuring of microdomain organization of lipids in bilayer. Melafen action on protein interactions logged on freshly-isolated and aged membranes of erythrocyte ghosts. In the process of ageing of interconnection of cytoskeleton components are decaying, and the effects manifestation active matter increase. For melafen it not been detected of destructed actions to the proteins microdomains. Melafen influence on whole cells - erythrocytes were tested when measured the spontaneous hemolysis and the hemolysis when extremely conditions - in hypo and hypertonic mediums by the spectral method. Another approach allowed the registration of caffeine influences on Са2+-pump activity - ATPase, and Са2+-канала in organelles - fragments of sarcoplasmic reticulum (SR). The functional properties of organelles were tested potentiometric when ATP hydrolysis. This methodical way is consisted in increasing of the SR calcium storage capacity. The short-time ageing of organelles raises the bilayer permeability for oxalate anions. The oxalate is entering to the SR lumen and links free Са2+ions. Constantly it is the lowering of the concentration of free Са2+ inside SR. The oxalate raises the ability to Са2+-accumulating. Thus the activating of the ATPase pumping so much higher than its physiological significance occurred. These results allude to the fact that the action BAS to structural and the functional properties of biologic objects may be so significant changed in vitro, when they are studying in special methodic conditions. Thus we may get the clear and available for registration bio-answers.

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