Involvement and relative importance of at least two distinct mechanisms in the effects of 2-mercaptoethanol on murine lymphocytes in culture.
2-Mercaptoethanol (2-ME) exerts several effects on murine lymphocytes in culture that might explain its ability to enhance survival and growth of these cells. The uptake of the essential amino acid cystine and consequently the maintenance of intracellular glutathione levels are enhanced by 2-ME. Furthermore, 2-ME (even in the disulfide form) causes lymphocytes to release thiols into the culture medium. These effects might protect the cells from oxidative damage. The additional cystine provided by treatment of lymphocyte cultures with 2-ME might also allow adequate protein synthesis to support survival and/or growth. This study was conducted to assess the relative importance of the antioxidant and protein synthesis effects of 2-ME. As expected, 2-ME increased cystine uptake at all concentrations that enhanced growth and survival, but four nonthiol antioxidants that enhanced growth and/or survival either did not substantially affect cystine uptake or decreased it and did not affect the release of cystine or its products. The results presented here demonstrate that antioxidant protection is necessary and sufficient for lymphocyte survival and that cystine uptake in untreated lymphocytes is sufficient to support the protein synthesis needed for survival and limited growth. However, we also noted that concentrations of 2-ME that stimulated maximal growth more than doubled protein synthesis as measured at 8 hr. Thus the portion of the effects of 2-ME not accounted for by antioxidant action could be accounted for by enhanced protein synthesis.
Department of Biological Sciences, Mississippi State University, Mississippi State 39762.
S B
SB
Pruett
N
N
Obiri
J L
JL
Kiel
D000818:Animals; D000975:Antioxidants; D001692:Biological Transport / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003553:Cystine / Q000378:metabolism; D000860:Hypoxia; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000187:drug effects*; D008623:Mercaptoethanol / Q000494:pharmacology*; D051379:Mice; D014176:Protein Biosynthesis
D000975:Antioxidants; D003553:Cystine; D008623:Mercaptoethanol