Glutathione status during the mitogenic response of rat splenocytes. Effects of oxygen concentration: FO2 21% versus FO2 7%

Glutathione is known to be an important parameter for ConA proliferative response of murine splenocytes. We studied the glutathione status of ConA-stimulated rat splenocytes during the early and late phase of the mitogenic response under low (FO2 7%) and standard (FO2 21%) oxygen concentrations. We determined the intracellular total, oxidized and reduced glutathione levels after 6, 12, 24 and 48 h of culture with or without ConA and/or 2-ME, under FO2 7% and 21%. Our results showed that: The 2 GSSG/GSH + 2 GSSG ratio, which indicated the redox state of the cells, remained normal during the early period of culture (0-24 h), irrespective of culture conditions. After 48 h of culture, this ratio increased dramatically under FO2 21% and less under FO2 7%. The maintenance of the redox state seems to be an oxygen concentration-dependent phenomenon. ConA stimulation involved a glutathione consumption during the early stages of culture; under these conditions 2-ME increased the glutathione synthesis, which was higher under FO2 7% than under FO2 21%. On the other hand, the presence of 2-ME involved an increase of tritiated thymidine uptake in stimulated splenocytes, which was significantly higher under FO2 21% than under FO2 7%. Low oxygen tension (FO2 7%) can induce a higher increase of glutathione synthesis, whereas the respective ConA proliferative response is lower than that observed under standard O2 conditions.