Comparison of the delivery of reduced glutathione into P388D1 cells by reduced glutathione and its mono- and diethyl ester derivatives.

The effect of reduced glutathione, reduced glutathione monoethyl ester and reduced glutathione diethyl ester on the cellular concentration of reduced glutathione and cysteine in P388D1 macrophages in vitro, and the cellular and extracellular de-esterification of reduced glutathione esters, was investigated. At 1 mM reduced glutathione derivative, only reduced glutathione diester markedly increased the cellular concentration of reduced glutathione. There was little delivery of reduced glutathione monoethyl ester into the cells. Reduced glutathione, and monoethyl and diethyl ester derivatives all increased the cellular concentration of cysteine; reduced glutathione diethyl ester also increased the cellular concentration of gamma-glutamylcysteine. Reduced glutathione diethyl ester also increased the cellular concentration of gamma-glutamylcysteine. Reduced glutathione esters were de-esterified intracellularly where the diester was rapidly converted to the monoester. The diester was also converted to the monoester extracellularly by interaction with cell surface esterases and by a much slower spontaneous hydrolysis. This indicates that the diester of reduced glutathione was a much more effective vehicle for delivery of reduced glutathione into cells than the monoester. Reduced glutathione diester also increased the cellular concentrations of cysteine and gamma-glutamylcysteine, suggesting that de novo synthesis of reduced glutathione was also stimulated.