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Injuries to cultivated BJA‐B cells by ajoene, a garlic‐derived natural compound: Cell viability, glutathione metabolism, and pools of acidic amino acids
Author(s) -
Scharfenberg Klaus,
Ryll Thomas,
Wagner Roland,
Wagner Karl G.
Publication year - 1994
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041580108
Subject(s) - glutathione , viability assay , trypan blue , biochemistry , chemistry , oxidative stress , cytotoxicity , cell , biology , in vitro , enzyme
Ajoene (4,5,9‐trithiadodeca‐1,6,11‐triene‐9‐oxide), a garlic‐derived natural compound, which had been shown to have cytostatic/cytotoxic properties, was tested with a B cell lymphoma‐derived cell line (BJA‐B cells) in order to elucidate its mechanism of cytotoxic action. Viability of the cells was determined by the Trypan blue exclusion test and the colorimetric tetrazolium (MTT) assay, whereas metabolic disturbance was evaluated by measuring the pools of reduced (GSH), oxidized glutathione (GSSG) and the acidic amino acids, Glu and Asp. Fast uptake of ajoene was accompanied by an immediate reduction of the CSH and increase in the GSSG levels. The extent of these changes, as well as the further development of the metabolite pools, depended on the ajoene dose per cell. At a sublethal ajoene dose the GSH and GSSG pools rose at the later stages to levels much higher than in the control experiment. Bleb formation at the cytoplasmic membrane was a further rapid phenomenon, although injuries detected by Trypan blue exclusion developed only at a later stage. The MTT assay, performed in a parallel experiment (48 h after ajoene addition), showed, however, that reduction of cell viability was established at the very beginning of ajoene exposure. Altogether, the action of ajoene strongly resembled oxidative stress (i.e., interference with SH homeostasis and its pleiotropic consequences to cell physiology and metabolism). © 1994 Wiley‐Liss, Inc.