Effects of alpha-phenyl N-tert-butylnitrone, a spin trap reagent, on the proliferation of murine hematopoietic progenitor cells in vitro.
We investigated the effect of alpha-phenyl N-tert-butylnitrone (PBN), a spin trap reagent, on the proliferation of murine hematopoietic progenitor cells in vitro. During the addition of PBN to the liquid cultures of murine bone marrow cells containing a combination of interleukin-3, interleukin-6 and the c-kit ligand/stem cell factor, colony-forming cells in vitro (CFC) and the colony-forming unit in the spleen (CFU-S) increased about 1.6-fold and 2.0-fold, respectively, higher than the control culture. These effects were not observed when using dimethyl sulfoxide, which has the ability to scavenge radicals, and 5,5-dimethyl-1-pyrroline N-oxide, another spin trap reagent. Analysis of cultured cells from a 7-day liquid culture with PBN revealed that the ratio of the intracellular glutathione (GSH) and GSH/GSSG (oxidized GSH) content was higher than the control. Adding thiol N-acetylcysteine, a thiol reagent and a precursor of intracellular GSH, also showed similar effects on the liquid culture of murine hematopoietic progenitor cells and the level of intracellular GSH. In contrast, adding DL-buthionine-[S,R]-sulfoximine, a gamma-glutamylcysteine synthetase inhibitor, decreased the intracellular GSH level and did not increase the number of CFC and CFU-S. These results suggest that PBN regulates the content of intracellular thiol molecules, and the possibility of a relationship between the intracellular redox state and the proliferation and differentiation of hematopoietic stem cells.
Kashiwakura
I
I
Laboratory of Radiopharmaceutical Sciences, Hokkaido College of Pharmacy, Otaru, Japan.
Kuwabara
M
M
Murakami
M
M
Hayase
Y
Y
Takagi
Y
Y
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D003497:Cyclic N-Oxides; D005978:Glutathione / Q000032:analysis; D019803:Glutathione Disulfide / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D007377:Interleukin-3 / Q000737:chemistry / Q000494:pharmacology; D015850:Interleukin-6 / Q000737:chemistry / Q000494:pharmacology; D008297:Male; D051379:Mice; D009589:Nitrogen Oxides / Q000494:pharmacology*; D013113:Spin Labels; D019089:Stem Cell Factor / Q000737:chemistry / Q000494:pharmacology
D003497:Cyclic N-Oxides; D007377:Interleukin-3; D015850:Interleukin-6; D009589:Nitrogen Oxides; D013113:Spin Labels; D019089:Stem Cell Factor; C029217:phenyl-N-tert-butylnitrone; D005978:Glutathione; D019803:Glutathione Disulfide; D000111:Acetylcysteine