Toxicological Sciences, Vol 49, 297-305, Copyright © 1999 by Society of Toxicology
K Haidara, P Moffatt and F Denizeau
Metallothionein (MT) is a protein involved in heavy metal homeostasis and
detoxification. According to several studies, MT could be involved in the
antioxidant defense system, in which glutathione (GSH) is an essential
component. The aim of this study was to verify the implication of MT in the
antioxidant defense system in isolated rat hepatocytes. For this purpose,
hepatocyte cultures were exposed to treatments known to modify MT or GSH
levels. Zinc (Zn) was used as an inducer of MT while diethyl maleate (DEM)
and buthionine sulfoximine (BSO) were used as GSH depletors. GSH, MT, and
antioxidant enzyme activities were measured under conditions of MT
induction and GSH depletion. Induction of MT synthesis through an 18-hour
exposure to Zn (20 microM), did not result in any significant change in GSH
levels or in activities of the antioxidant enzymes, glutathione-peroxidase
(GSH- Px), catalase, and superoxide dismutase (SOD). DEM caused GSH
depletion in cells, whether they were exposed to Zn or not, that lasted one
h; after that time, GSH rose back to basal levels. BSO also caused GSH-
depletion in cells exposed or unexposed to Zn, and no recovery in GSH
levels was detectable during the entire period of exposure (12 h). However,
GSH depletion induced by both DEM or BSO was attenuated in Zn- treated
hepatocytes. Moreover, DEM and BSO exposures led to a depletion of MT
levels in Zn-treated hepatocytes, indicating a link between GSH and MT
metabolism. In cells unexposed to either Zn, DEM or BSO, there was an
increase in GSH-Px and SOD activities after 6 and 12 h of incubation,
respectively. Under the same conditions, catalase activity was inhibited
after 6 h of incubation and returned to the activity found at t = 0 after
12 h of incubation. DEM and BSO treatments had no significant effect on
GSH-Px or SOD activities although they led to inhibition of catalase
activity. Taken together, our data indicate that MT induction, which
creates a new pool of thiol groups in the cell cytosol, can attenuate GSH
depletion induced by DEM or BSO. It appears that catalase is most sensitive
to oxidative stress and that MT induction can antagonize the deleterious
effects of such stress on the enzyme. This study supports the view that MT
is part of the hepatocyte antioxidant-defense-system.
ARTICLES
Metallothionein induction attenuates the effects of glutathione depletors in rat hepatocytes
Departement de Chimie, Universite du Quebec a Montreal, Quebec, Canada.
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