Kinetic parameters of respiration in rat permeabilized hepatocytes upon Са2+ in various concentrations in medium and prolonged Influence of taurine

V. M. Merlavsky,R. Ostapiv,O. Ikkert,V. Manko,Ukraine Feed Additives

Published 2015 in Unknown venue

ABSTRACT

The dependence of permeabilized rat hepatocytes respiration rate on oxidation sub­ strates concentration at presence 0.1 and 1 μM Са2+ in the medium and amino sulfonic acid – taurine – effect on this subordination was studied. Experimental animals were in­ jected with taurine (40 mg/kg of weight) for 28 days and control rats were injected with equal volume of water. Liver cells were permeabilized with digitonin (20 μg per 1 million cells). Respiration rate was determined by polarographic method using Clark’s electrode, upon either succinate or pyruvate with malate oxidation. Hill equation parameters were calculated applying {v; v/[S]h} coordinates. The kinetic dependence of respiration upon succinate and pyruvate oxidation is well described by Hill equation in the mediums with studied concentration of Са2+ in control and under taurine action. Hill coefficient h and semi activation constant K0.5 for succinate, at rotenone presence, are not changed in cont­ rol animals after increase of Са2+ сoncentration from 0.1 to 1 μM. Maximal velocity Vmax was slightly increased under such conditions. Kinetic parameters of respiration upon suc­ cinate oxidation and both Са2+ concentration, are not changed significantly under pro­ longed taurine injection. There is more essential difference between kinetic parameters of pyruvate­stimulated respiration at various Ca2+ concentrations. Thus, in control, Vmax is 1.5 times lower, and K0.5 – 10.7 times major at 0.1 μM Са2+ in comparison with 1 μM Са2+. Hill coefficient becomes less than 1 at both Са2+concentration. As a result of taurine injection, Vmax is 1.6 times less at low Са2+ and K0.5 is higher, but only 6 times. Hill coefficient of this process increases at 0.1 μM Са2+ and is 1.12, upon taurine impact. Substrate inhibition, inherent to the dependence of respiration rate on pyruvate concentration within the me­ dium with 1 μM Са2+, starts developing, under taurine effect, at higher concentration of this substrate in comparison with control (3 mM comparatively to 0.35 mM). Thus, Са2+ con­ centration rise from 0.1 to 1 μM stimulates the rat hepatocytes mitochondria respiration upon exogenous pyruvate oxidation, but not succinate. Taurine under the prolonged in vivo action does not influence succinate-stimulated rotenone-insensitive respiration and intensifies processes of pyruvate-stimulated oxygen consumption in hepatocytes.

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