Involvement of glutathione peroxidase and catalase in the disposal of exogenous hydrogen peroxide by cultured astroglial cells.
The ability of astroglial cells to detoxify exogenously applied hydrogen peroxide (H2O2) was tested using astroglia-rich primary cultures derived from the brains of newborn rats. Incubation of astroglial cells with 100 microM H2O2 in the absence of glucose led to a 66% oxidation of the cellular glutathione within 30 s. Under these conditions, the cells were unable to re-establish the original high ratio of GSH/GSSG within 30 min of incubation. In contrast, if glucose was present the amount of GSSG produced on incubation with H2O2 was smaller (45% of total glutathione after 30 s) and the original ratio of GSH/GSSG was almost completely re-established within 10 min. If 100 microM H2O2 was applied, H2O2 disappeared from the incubation buffer with an apparent half-life of approximately 4 min. After 15 min of incubation, no H2O2 was detectable any more. The apparent half-life of H2O2 in the incubation buffer increased slightly but significantly with increasing concentration of H2O2 or when the cells were starved of glucose. A small reduction in the capacity of the cells to detoxify H2O2 was also observed after depletion of the glutathione content to 14% of control level by a 24 h pre-incubation of the cells in culture medium containing buthionine sulfoximine, an inhibitor of glutathione synthesis. Incubation of astroglial cells with mercaptosuccinate or 3-aminotriazole, inhibitors of glutathione peroxidase and catalase, respectively, only marginally reduced the rate of disappearance of H2O2 from the incubation buffer. In contrast, the rate of H2O2 clearance was strongly reduced in the presence of both inhibitors. These results demonstrate that glutathione peroxidase and catalase are involved in the detoxification of H2O2 by astroglial cells and that both enzymes are able to substitute for each other in the detoxification of H2O2.
Dringen
R
R
Physiologisch-Chemisches Institut, Eberhard-Karls-Universität Tübingen, Germany. ralf.dringen@uni-tuebingen.de
Hamprecht
B
B
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000378:metabolism*; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002374:Catalase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000378:metabolism; D019803:Glutathione Disulfide; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006207:Half-Life; D006861:Hydrogen Peroxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D017208:Rats, Wistar
D004791:Enzyme Inhibitors; D019328:Buthionine Sulfoximine; D006861:Hydrogen Peroxide; D002374:Catalase; D005979:Glutathione Peroxidase; D005978:Glutathione; D005947:Glucose; D019803:Glutathione Disulfide