L-Cysteine influx and efflux: A possible role for red blood cells in regulation of redox status of the plasma

The objective of this study was to investigate if erythrocytes play a role in the maintenance of redox homeostasis of the plasma. Thus, we studied l-cysteine efflux and influx in vitro in human erythrocytes. In the present study, we exposed the erythrocytes to different concentrations of l-cysteine and then measured the intracellular free -SH concentrations. Erythrocytes treated in the same manner were later utilized for the cysteine efflux studies. The effect of temperature on the influx and the efflux processes were also evaluated. Change in the free -SH content of the buffer was evaluated as a measure for the presence of an efflux process. The effects of free -SH depletion on l-cysteine transport is also investigated. We also determined the rate of l-cysteine efflux in the presence and absence of buthionine sulfoximine (BSO) in erythrocytes that are pretreated with 1-chloro-2,4-dinitro benzene, a glutathione (GSH) depletory. Our l-cysteine influx studies demonstrated that erythrocytes can respond to increases in l-cysteine concentration in the extracellular media and influx l-cysteine in a concentration-dependent manner. Free -SH concentrations in erythrocytes treated with 1 mM l-cysteine reached to 1.64 0.06 mM in 1 h whereas this concentration reached to 4.30 0.01 mM in 10 mM l-cysteine treated erythrocytes. The l-cysteine efflux is also determined to be time-and concentration-dependent. Erythrocytes that are pretreated with higher l-cysteine concentrations displayed a higher efflux process. Outside concentration of free -SH in 1 mM l-cysteine pretreated erythrocytes reached to 0.200 0.005 mM in 1 h whereas this concentration reached to 1.014 0.002 with 10 mM l-cysteine pretreated erythrocytes. Our results also indicate that the rate of inward and outward transport of l-cysteine is affected by the oxidative status of the erythrocytes. When GSH is depleted and GSH synthesis is blocked, the l-cysteine uptake and the efflux processes are significantly decreased. Depending on our results, it could be concluded that erythrocytes play a role in the regulation of the plasma redox status and intracellular level of GSH determines the rate of the l-cysteine efflux.