The hemolytic effect of copper complexes with different ligands.

The effect of Cu(II) and Cu(II) complexes/Cu(II)(cimetidine)2 Cu(II)(glycine) and Cu(II)(glycine)2/on the rat and human erythrocytes was studied. Time- and dose-dependence of the Cu(II) complexes-induced hemolysis, oxidation of hemoglobin and formation of precipitates of denaturated hemoglobin were established. The hemolytic effect of Cu(II)complexes was abolished by strong complexons of Cu(II) and by physiological concentrations of albumin and histidine. The hemolysis greatly decreased in the presence of chelator of Cu(I) and superoxide dismutase, which suggested oxi-reduction transitions and participation of superoxide in the process. The results are considered with the view to clarifying the differences in the stability and reaction ability of the different complexes, the possibility of their parenteral application.

A study on the ability of copper complexes to act as active oxygen species scavengers.

The ability of two low-molecular-weight copper complexes to influence the hemolysis of human erythrocytes caused by active oxygen species-generating systems was studied. Cu(II) (glycine)2 and Cu(II) (tyrosine)2 did not inhibit hemolysis due to O-2 and H2O2 generated by xanthine oxidase plus acetaldehyde but rather has a prooxidant effect. The same copper complexes as well as Cu(II) strongly inhibited the hemolysis caused by the 1O2-generating system (Rose Bengal + light). It was found that except for 1O2 the other active oxygen species (O-2, H2O2 and OH.) did not participate in the Rose Bengal + light-induced hemolysis. Thus we examined whether the inhibitory effect of copper complexes was due to 1O2 quenching. Cu(II) (glycine)2 inhibited the Rose Bengal + light-induced oxidation of compounds known to react chemically with 1O2 and its effects were analogous to the effects of physical 1O2 quenchers, e. g. NaN3 and NiCl2. The oxygen consumption upon NADH-photooxidation in the presence of Rose Bengal was inhibited competitively by Cu(II) (glycine)2 but when concentration of Rose Bengal or light intensity were varied the extent of Cu(II) (glycine)2-caused inhibition was not changed. It is concluded that the effects of Cu(II) (glycine)2 and possibly of Cu(II) (tyrosine)2 are due to quenching of 1O2 but quenching of the excited state of the dye could not be excluded.

Rose bengal-induced photodamage to subcellular membranes and enzymes.

Studies were made of photodamage (photolysis) caused by Rose Bengal + light of peroxisomes, mitochondria, and lysosomes, as well as inhibition of the respective marker enzymes: catalase, sulfite oxidase, and acid phosphatase. The time-dependence of lipid peroxidation caused by the same system and measured by the increase of thiobarbituric acid-reactive substances was also examined. The most sensitive to photolysis were the peroxisomes and the least susceptible were lysosomes as the inhibition of their marker enzymes followed this order. The sensitivity of the subcellular membranes to digitonin (a sterol binding agent) showed the opposite dependence. The importance of the cholesterol content for the membrane stability to active oxygen species, as well as the role of lipid peroxidation in photodynamic damage of the systems studied, is discussed.

Heterogeneity of copper-induced superoxide dismutase in rat liver lysosomes.

Preparations of liver large granule fraction from control and copper-injected rats were treated with digitonin (0.01-0.16 mg/mg protein) and supernatants rich in lysosomal matrix and in mitochondrial intermembrane space were obtained. After copper injection the superoxide dismutase activity in all supernatants was significantly increased. The granular CuZn-superoxide dismutase in the two animal groups was localized in lysosomes only. The cytosolic and lysosomal CuZn-superoxide dismutase in control preparations showed an equal electrophoretic pattern (two peaks with Rf = 0.42 and 0.47). After copper injection three new electrophoretic peaks of the lysosomal CuZn-superoxide dismutase activity (with Rf = 0.37, 0.52 and 0.62 respectively) appeared. The increased heterogeneity of the granular CuZn-superoxide dismutase activity after copper injection is explained by the oxidative degradation of the CuZn-superoxide dismutase in lysosomes.

[Effect of chlorpromazine, lidocaine and procaine on the electrophoretic mobility of rat erythrocytes]. / Vliianie na khlorpromazin, lidokain i prokain vûrkhu elektroforetichnata podvizhnost na eritrotsiti ot plûkh.

The authors examined the influence of chlorpromazine on the electrophoretic motility of erythrocytes by means of the method of microelectrophoresis. The results were compared with data, obtained from analogous studies on the effect of local anaesthetics (procaine and lidocaine) on erythrocyte membranes. Erythrocytes (6.10(8) erythrocytes/ml) were incubated at temperature of 20 degrees C for a period of 15 minutes. They found that erythrocytes diminished their electrophoretic motility after incubation with labilizing concentration (10(-5M) of chlorpromazine as well as of procaine and lidocaine, but after treatment with stabilizing concentration (10-7M) they increased their electrophoretic motility. The obtained results showed that there was a reverse dependence between the hemolytic and pharmacological action of these three preparations and the change, which they induced on the superficial electrical charge-a greater change in the superficial electric charge induced those, which were weaker hemolytics and had weaker pharmacological effect.