EFFECTS OF BIOGENIC AND ABIOGENIC DISULPHIDES UPON ENDOTHELIAL CELLS IN CULTURE: COMPARISON OF THREE METHODS OF VIABILITY ASSESSMENT
D.S. Prokofieva,1,* N.V. Goncharov 1,2
1 Research Institute of Hygiene, Occupational Pathology and Human Ecology and
2 I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry RAS, St. Petersburg;
* e-mail: darija-p1@yandex.ru
Effects of biogenic and abiogenic disulphides on viability of human umbilical vein endothelial cells in culture has been investigated using three methods: the neutral red uptake assay,
quantification of intracellular ATP, and modifications of Mosmann method, the essence of which is the reduction of tetrazolium salts, MTT and MTS, by cells. 2,2R-dithio-bis(N,N-diethyl)
ethanamine (DS) was used as an abiogenic disulphide. As for biogenic disulphides, we used GSSG and garlic oil (GO), the principal component of which is diallyl disul-phide (DADS).
It has been found that DS and GO have a similar cytotoxic effect upon the endothelial cells (EC50 ~ 0.6 mM). GSSG in concentrations up to 1 mM did not effect the viability
of endothelial cells. It has been demonstrated for the first time that DS and GO can serve as mediators of plasma membrane oxidoreductase activity, tetrazolium salts being as the substrate;
this may cause false-negative effect. Thus, the Mosmann method has serious limitations when testing the cytotoxicity of disulphides, though can be used in studying the mechanism of action
of disulphides.
Key words: disulfides, glutathione, garlic oil, endothelium, cytotoxicity
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