Changes In GSH Levels Research Articles

Age and sex differences in human skeletal muscle: Role of reactive oxygen species

Previous studies, conducted on experimental animals, have indicated that reactive oxygen species (ROS) are involved in the aging process. The objective of this work was to study the relationship between oxidative damage and human skeletal muscle aging, measuring the activity of the main antioxidant enzymes superoxide dismutase (total and MnSOD), glutathione peroxidase (GPx) and catalase in the skeletal muscle of men and women in the age groups: young (17–40 years), adult (41–65 years) and aged (66–91 years). We also measured glutathione and glutathione disulfide (GSH and GSSG) levels and the redox index; lipid peroxidation and protein carbonyl content. Total SOD activity was lower in the 66–91 year-old vs. the 17–40 year-old men; MnSOD activity was significantly greater in 66–91 year-old vs. 17–40 year-old women. GPx activity remained unchanged. The activity of catalase was lower in adults than in young men but higher in the aged. We observed no changes in GSH levels and significantly higher GSSG levels only in aged men vs. adult men, and a significant decrease in aged women vs. aged men. The protein carbonyl content increased significantly in the 41–65 and 66–91 year-old vs. the 17–40 year-old men. Finally, young women have lower lipid peroxidation levels than young men. Significantly higher lipid peroxidation levels were observed in aged men vs. both young and adult men, and the same trend was noticed for women. We conclude that oxidative damage may play a crucial role in the decline of functional activity in human skeletal muscle with normal aging in both sexes; and that men appear to be more subject to oxidative stress than women.

Antioxidant administration inhibits exercise-induced thymocyte apoptosis in rats.

The purpose of this study was to investigate the effect of antioxidant on exercise-induced apoptosis in rat thymocytes. After exercise at 13.8 m x min(-1) for 60-90 min x d(-1) on a motor-driven drum exerciser for 2 consecutive days, rat thymocyte apoptosis was monitored by the feature of DNA fragmentation. To study the effect of antioxidant, rats were administered with butylated hydroxyanisole (BHA) for 7 d before exercise. Exercise could induce thymocyte DNA fragmentation as detected on electrophoretic gel and by cell death detection ELISA kit. Further studies indicated that pretreatment with antioxidant BHA to rats resulted in a blockage of exercise-induced DNA fragmentation. The concentrations of glutathione (GSH) were not significantly changed in rat thymocytes after exercise with or without BHA treatment. These results suggest that reactive oxygen species may play a role in thymocyte apoptosis induced by exercise. However, changes in GSH levels were not observed in this exercise model.

Phospholipase D activation in endothelial cells is redox sensitive.

Reactive oxygen species (ROS) are implicated in the pathophysiology of a number of vascular disorders, including atherosclerosis. Recent studies indicate that ROS modulate signal transduction in mammalian cells. Previously, we have shown that ROS (hydrogen peroxide, fatty acid hydroperoxide, diperoxovanadate, and 4-hydroxynonenal) enhance protein tyrosine phosphorylation and activate phospholipase D (PLD) in bovine pulmonary artery endothelial cells (BPAECs). In the present study, our aim was to investigate the role of exogenous thiol agents on ROS-induced PLD activation in conjunction with the role of cellular thiols--glutathione (GSH) and protein thiols--on PLD activation and protein tyrosine phosphorylation. Pretreatment of BPAECs with N-acetyl-L-cysteine (NAC) or 2-mercaptopropionylglycine (MPG) blocked ROS-induced changes in intracellular GSH and PLD activation. Also, pretreatment with NAC attenuated diperoxovanadate-induced protein tyrosine phosphorylation. Pretreatment of BPAECs with diamide or L-buthionine-(S,R)-sulfoximine (BSO), agents that lower intracellular GSH and thiols, enhanced PLD activity. Furthermore, NAC blocked diamide- or BSO-mediated changes in GSH levels, PLD activity, and protein tyrosine phosphorylation. NAC also attenuated diamide-induced tyrosine phosphorylation of proteins between 69 and 118 KDa. These results support the hypothesis that modulation of thiol-redox status (cellular nonprotein and protein thiols) may contribute to the regulation of ROS-induced protein tyrosine phosphorylation and PLD activation in vascular endothelium.

Metallothionein induction attenuates the effects of glutathione depletors in rat hepatocytes.

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.

Induction of cystine transport via system x c− and maintenance of intracellular glutathione levels in pancreatic acinar and islet cell lines

The relationship between L-cystine transport and intracellular glutathione (GSH) levels was investigated in cultured pancreatic AR42J acinar and βTC3 islet cells exposed to diethylmaleate, an electrophilic agent known to activate cellular antioxidant responses. Cystine transport was mediated predominantly by the Na +-independent anionic amino acid transport system x c −, with influx inhibited potently by glutamate and homocysteate but unaffected by cationic or neutral amino acids. Saturable cystine transport was 10-fold higher in AR42J (531 pmol (mg protein) −1 min −1) than in βTC3 (49 pmol (mg protein) −1 min −1) cells, and GSH levels were higher in AR42J cells. Treatment with 2-mercaptoethanol increased GSH levels in βTC3 cells from 7.5 to 36 nmol (mg protein) −, whilst the GSH content in AR42J cells (64 nmol (mg protein) −1) was not altered significantly. Incubation of AR42J or βTC3 cells with homocysteate (2.5 mM, 0–48 h), a competitive inhibitor of cystine transport via system x c −, reduced intracellular GSH levels and resulted in a time-dependent (6–24 h) induction of system x c − transport activity. Treatment of AR42J cells with diethylmaleate (100 μM 0–48 h) resulted in a time- (5–10 h) and protein synthesis-dependent induction of cystine transport, with intracellular GSH levels initially decreasing and then increasing 2-fold above control levels after 24 h. Diethylmaleate also depressed GSH levels in βTC3 cells, but cystine transport was not elevated significantly. In both AR42J and βTC3 cells, inhibition of γ-glutamyl cysteine synthetase by buthionine sulphoximine (100 μM, 24 h) reduced GSH levels but had no effect on cystine transport. The present findings establish that induction of system x c − leads to changes in GSH levels in pancreatic AR42J acinar and βTC3 islet cells, with changes in the intracellular redox state stimulating transporter expression. Induction of activity of system x c −, together with adaptive increases in GSH synthesis in response to oxidative stress, may contribute to cellular antioxidant defences in pancreatic disease.

Glutathione Regulation of Neutral Sphingomyelinase in Tumor Necrosis Factor-α-induced Cell Death

Tumor necrosis factor-alpha (TNFalpha)-induced cell death involves a diverse array of mediators and regulators including proteases, reactive oxygen species, the sphingolipid ceramide, and Bcl-2. It is not known, however, if and how these components are connected. We have previously reported that GSH inhibits, in vitro, the neutral magnesium-dependent sphingomyelinase (N-SMase) from Molt-4 leukemia cells. In this study, GSH was found to reversibly inhibit the N-SMase from human mammary carcinoma MCF7 cells. Treatment of MCF7 cells with TNFalpha induced a marked decrease in the level of cellular GSH, which was accompanied by hydrolysis of sphingomyelin and generation of ceramide. Pretreatment of cells with GSH, GSH-methylester, or N-acetylcysteine, a precursor of GSH biosynthesis, inhibited the TNFalpha-induced sphingomyelin hydrolysis and ceramide generation as well as cell death. Furthermore, no significant changes in GSH levels were observed in MCF7 cells treated with either bacterial SMase or ceramide, and GSH did not protect cells from death induced by ceramide. Taken together, these results show that GSH depletion occurs upstream of activation of N-SMase in the TNFalpha signaling pathway. TNFalpha has been shown to activate at least two groups of caspases involved in the initiation and "execution" phases of apoptosis. Therefore, additional studies were conducted to determine the relationship of GSH and the death proteases. Evidence is provided to demonstrate that depletion of GSH is dependent on activity of interleukin-1beta-converting enzyme-like proteases but is upstream of the site of action of Bcl-2 and of the execution phase caspases. Taken together, these studies demonstrate a critical role for GSH in TNFalpha action and in connecting major components in the pathways leading to cell death.

Optimal NFκB Mediated Transcriptional Responses in Jurkat T Cells Exposed to Oxidative Stress Are Dependent on Intracellular Glutathione and Costimulatory Signals

Many early response genes induced by the exposure of mammalian cells to environmental stress contain DNA sequences which bind nuclear factor (NF) κB. However, the effects of oxidative stress on NFκB activity in T cells are contradictory with evidence supporting both stimulation and suppression. The present investigation examined the effects of low levels of oxidative stress in the form of H2O2on NFκB transactivation in Jurkat T cells and the regulation of NFκB activity by cellular glutathione (GSH) levels and costimulatory signals. Transient transfection analyses demonstrated 2-3 fold increases in transcription of an NFκB dependent chloramphenicol acetyl transferase (CAT) reporter after the exposure of Jurkat cells to 100-500 μM H2O2. By comparison, dual stimulation of Jurkat cells with phytohemagglutinin (PHA) plus phorbol (PMA) induced 9-10 fold increases in NFκB CAT activity. Although no marked changes in GSH levels were detected in cells treated with H2O2or PHA/PMA, the depletion of GSH in cells pretreated withDL-buthionine-[S,R]-sulfoximine (BSO) substantially inhibited NFκB transactivation by H2O2. In addition, H2O2was less effective than PHA/PMA in inducing NFκB1 (p50)/RelA (p65) complexes. However, signals induced by oxidative conditions effectively cooperated with stimulatory signals provided by PMA but not with T cell receptor/CD3 stimulation to induce significant increases in NFκB CAT responses. These results suggest that a functional GSH system is required for NFκB activation in T cells exposed to oxidative reagents and provide evidence that oxidative stress triggers signaling events capable of participating with distinct coregulatory signals to activate NFκB transcriptional responses.

Hydrogen peroxide increases the activity of gamma-glutamylcysteine synthetase in cultured Chinese hamster V79 cells.

Hydrogen peroxide (H2O2) caused a rapid and a concentration-dependent increase in the activity of gamma-glutamylcysteine synthetase (gamma-GCS) in cultured Chinese hamster V79 cells. The increase in the activity was transient and declined rapidly during post-treatment incubation. Inhibition of protein synthesis by cycloheximide, chelation of divalent iron by o-phenanthroline, and scavenging of free radicals by butyl-4-hydroxyanisole failed to suppress the increase in activity of gamma-GCS caused by H2O2. However, catalase completely inhibited the increase in the activity of the enzyme. H2O2 did not change the level of total glutathione (GSH + GSSG) but it oxidized GSH. The increase in levels of GSSG caused by H2O2 was enhanced by o-phenanthroline. These results suggest that the increase in activity of gamma-GCS caused by H2O2 is not an inducible phenomenon, nor it is attributable to the action of free radicals generated by an iron-catalyzed Fenton reaction. Furthermore, the changes in levels of GSH and GSSG caused by H2O2 appear not to be responsible for the increase in activity of gamma-GCS caused by the hydroperoxide. However, chemical reduction of the enzyme, the activity of which had been increased by H2O2, resulted in a decrease in tha activity, suggesting the involvement of oxidation of the enzyme in the increased activity of gamma-GCS caused by H2O2. The results also suggest that the activity of gamma-GCS in cultured V79 cells can be regulated by the cellular oxidation-reduction state.

The effect of the pyrrolizidine alkaloids, monocrotaline and trichodesmine, on tissue pyrrole binding and glutathione metabolism in the rat

One day after in vivo administration of equitoxic doses of the hepatotoxic and pneumotoxic pyrrolizidine alkaloid, monocrotaline (65 mg/kg, i.p.) or the related hepatotoxic and neurotoxic alkaloid trichodesmine (15 mg/kg, i.p.) hepatic GSH levels are increased by more than 50%. These doses of alkaloids represent 60% of the ld 50 values. Accompanying these changes in GSH levels is an increase in the overall rate of GSH synthesis in supernatants of alkaloid-exposed livers. The ability of the rat to metabolize the two alkaloids was shown by the appearance of tissue-bound pyrrolic metabolites of pyrrolizidines in various organs. The levels of these metabolites appear to correlate with organ toxicity. For the hepatic and pneumotoxic alkaloid, monocrotaline, higher levels are found in liver (17 nmoles/g tissue) and lung (10 nmoles/g) than for trichodesmine (7 nmoles/g and 8 nmoles/g, respectively). For the neurotoxic alkaloid, trichodesmine, higher levels are found in brain (3.8 nmoles/g tissue) than for monocrotaline (1.7 nmoles/g tissue).

Glutathione levels of the human crystalline lens in aging and its antioxidant effect against the oxidation of lens proteins.

This paper reports the role of glutathione (GSH) in the crystalline lens as an antioxidant against the oxidation of lens protein. GSH levels in normal lenses decreased gradually with increasing age, from approximately 5 mumol per g lens (wet weight) to 3 mumol per g lens (wet weight). On the other hand, levels of oxidized GSH in the lenses increased until the age of 40. After that, it remained almost constant at the level of approximately 0.9 mumol per g lens. Protein-bound GSH levels in both soluble and insoluble lens proteins dropped noticeably in the 50-year and older age groups, although there were significant differences in levels between both fractions. A decrease of tryptophan and tyrosine residues in lens proteins was proportional to a decrease in GSH levels in the lens as a result of aging. Those residue levels in the cataractous lenses were approximately half those in the normal lens proteins, and GSH levels in such lenses were almost one-tenth that in the normal lens. This study revealed that GSH may play an important role in preventing the oxidation of lens proteins from various oxidants. Furthermore, it is conceivable that these normal changes in GSH levels in the lenses increase the vulnerability of the lens to senile cataractogenesis.

Effect of 2-mercaptoethanol on glutathione levels, cystine uptake and insulin secretion in insulin-secreting cells.

The role of glutathione (GSH) in the differentiated state of insulin-secreting cells was studied using 2-mercaptoethanol as a means of varying intracellular GSH levels. 2-Mercaptoethanol (50 microM) caused a marked increase of GSH in two rat insulinoma cell lines, RINm5F and INS-1, the latter being dependent on the presence of 2-mercaptoethanol for survival in tissue culture. The effect of 2-mercaptoethanol on GSH was shared by other thiol compounds. Since in other cell types 2-mercaptoethanol is thought to act on cystine transport, thereby increasing the supply of cysteine for GSH synthesis, we have studied [35S]cystine-uptake in INS-1 cells. At equimolar concentrations to cystine, 2-mercaptoethanol caused stimulation of [35S]cystine-uptake. The effect persisted in the absence of extracellular Na+, probably suggesting the involvement of the Xc- carrier system. INS-1 cells with a high GSH level, cultured 48 h with 2-mercaptoethanol, displayed a lower cystine uptake than control cells with a low GSH content. The effect of variations of the GSH levels on short-term insulin release was studied. No alteration of glyceraldehyde-induced or KCl-induced insulin release in RINm5F cells was detected. In contrast, both in islets and in INS-1 cells, a high GSH level was associated with a slightly lower insulin release. In INS-1 cells the effect was more marked at low glucose concentrations, resulting in an improved stimulation of insulin secretion. On the other hand, in islets, a decrease in the incremental insulin release evoked by glucose was seen. As in other cell types, oxidized glutathione (GSSG) was less than 5% of total GSH, and in INS-1 cells no change in the GSH/GSSG ratio was detected during glucose-induced or 3-isobutyl-1-methylxanthine-induced insulin release. In conclusion, 2-mercaptoethanol-dependent INS-1 cells, as well as RINm5F cells and islets of Langerhans, display a low capacity in maintaining intracellular levels of GSH in tissue culture without extracellular thiol supplementation; 2-mercaptoethanol possibly acts by promoting cyst(e)ine transport; changes in GSH levels caused a moderate effect on the differentiated function of insulin-secreting cells.

Glutathione release by pulmonary alveolar macrophages in response to particles in vitro

We studied whether cultured rat pulmonary alveolar macrophages (PAM) could release glutathione (GSH) in response to latex beads, quartz, and crocidolite asbestos. PAM were exposed for 2 h to 0–100 μg of particles per l × 10 6 cells. Both quartz and asbestos produced concentration-dependent increases (up to 8–10-fold) in the amount of GSH recovered in the medium and decreases in the cellular GSH levels. In contrast, latex beads did not produce any changes in GSH levels. We also measured lactate dehydrogenase (LDH) levels as an index of toxicity. Only quartz and asbestos were able to produce increases in LDH levels in the medium. The release of GSH occurred at particle concentrations that did not cause release of LDH. Our results indicated that, in vitro, PAM release GSH in response to toxic particles.

Erythrocyte GSH, hemoglobin and potassium concentrations during the postnatal period in Granadina goats

Variations in GSH level, hemoglobin and erythrocyte potassium were studied in 12 adult Granadina goats and their 15 female progeny during the first 30 postnatal weeks. Significantly ( P<0.01) lower mean values than in adults were detected from the 18th to the 25th week for GSH level. In this same period, the hemoglobin concentration was also lower ( P<0.01) than in adults. However, no correlation was detected between changes in GSH level and in potassium type (high vs. low concentration). Eight adult animals (66.7%) were of the high potassium (HK) type (41.0–63.0 mEq/l) and four were of the low potassium (LK) type (6.5–18.0 mEq/l). The switch from HK to LK type in kids took place at the 7th week postpartum. The mode of inheritance of HK and LK types supported the theory that the LK gene is dominant over the HK gene.

Renal and hepatic glutathione pool modifications in response to depletion treatments.

In this study we examined the response of the renal and hepatic glutathione (GSH) pool in rats to drastic GSH depletion treatments. For this purpose, we used a protein-free diet, starvation, and the injection of varying doses of diethyl maleate as depleting agents. We analysed GSH levels in both kidney and liver tissue homogenates after rats were fed a protein-free diet for 2 or 7 days or starved for 1, 2, or 3 days, as well as after diethyl maleate administration in a single maximal dose or in varying doses. The results indicated that the liver GSH pool was always more labile than the kidney GSH pool. Moreover, kidney GSH levels were almost unchanged after 7 days on a protein-free diet or after 2 days of starvation, while liver showed significant changes in GSH levels. When we analysed the repletion rate, kidney had higher kinetic parameters (k = 0.148 h-1) than liver (0.097 h-1). We conclude that efficient mechanisms of maintaining GSH levels exist in the kidney and these may serve to avoid GSH diminution and hence preserve renal function during states of GSH depletion.

Reduction in erythrocytic GSH level and stability in Plasmodium vivax malaria

The existence of haemolytic anaemia in malaria indicates disturbances in red cell stability due to physical as well as metabolic stress attributable to the malarial parasite. As erythrocytic reduced glutathione (GSH) is involved in maintaining the integrity of red cells, the status of erythrocytic GSH was studied in 40 patients infected with Plasmodium vivax before and after therapy with chloroquine. 40 normal subjects, age- and sex-matched, were studied as controls. The level of erythrocytic GSH of malaria patients during infection and before therapy was significantly lower in comparison with controls (P < 0·0005). Instability of GSH was recorded in 17 of 40 patients, while none of the controls showed such a defect. There was a progressive decrease in GSH level and stability of the host red cells with increasing parasitaemia. Normal values were obtained following therapy and cure of malaria indicating that the changes in GSH level and stability are induced by P. vivax. Alterations in the GSH metabolism may represent one of the factors contributing to the severity of anaemia in malaria due to P. vivax infection.

The effect of methyl oleate hydroperoxide, a possible toxic ozone intermediate, on human normal and glucose-6-phosphate dehydrogenase-deficient erythrocytes

Erythrocytes of both normal and glucose-6-phosphate dehydrogenase (G-6-PD)-deficient humans responded in a dose-dependent manner to the oxidant stress of methyl oleate hydroperoxide (MOHP) as measured by decreases in G-6-PD activity, increases in methemoglobin (METHB) levels, and decreases in reduced glutathione (GSH). The G-6-PD-deficient erythrocytes displayed a markedly enhanced sensitivity to MOHP-induced decreases in G-6-PD activity and METHB increases while being less sensitive than normal erythrocytes to changes in GSH levels.

The effect of methyl linoleate hydroperoxide (MLHP), a possible toxic intermediate of ozone, on human normal and glucose‐6‐phosphate dehydrogenase (G‐6‐PD) deficient erythrocytes

Abstract Erythrocytes of both normal and G‐6‐PD deficient humans responded in a dose‐dependent manner to the oxidant stress of MLHP as measured by decreases in G‐6‐PD activity, increases in methemoglobin (METHB) levels and decreases in reduced glutathione (GSH). The G‐6‐PD deficient eryth‐rocytes displayed a markedly enhanced sensitivity to MLPH induced decreases in G‐6‐PD activity and METHB increases while being less sensitive than normal erythrocytes to changes in GSH levels.

Alterations in tissue glutathione and angiotensin converting enzyme due to inhalation of diesel engine exhaust.

Quantitative changes in reduced glutathione (GSH) and angiotensin converting enzyme (ACE) of lung and extrapulmonary tissues were determined following exposure of laboratory animals to diesel engine exhaust (DEE). Exposure of male rats and guinea pigs to DEE containing 750 micrograms particulates per cubic meter for 12 wk did not cause any changes in GSH levels of lung, liver, and heart compared to control values. Rats were then exposed for various time periods to 6 mg/m3 DEE. Two weeks of exposure produced statistically significant increases of 21 and 7% in GSH levels of lung and liver, respectively, but not change in the heart. Following 4 wk of exposure, lung showed a 14% increase and heart an 11% increase in GSH level. Furthermore, rats exposed for 4 wk to DEE did not show any particular susceptibility toward the GSH-depleting effect of acetaminophen as compared to controls. A significant depletion (15%) of hepatic GSH was observed after 8 wk of exposure, while lung was still showing an increase of 18% in GSH and heart was unaffected. Time-dependent increases of 18 and 33% in serum ACE activity were noted after 4 and 8 wk of exposure. Pulmonary ACE activity did not decrease until after 8 wk, and then to a small extent (7%). The observed increases in GSH may have been related to the presence of NO2 in the DEE. On the other hand, the depletion of hepatic GSH suggests production of electrophilic compounds due to an induction of metabolic activity of liver. The change in serum ACE activity may be due to time-requiring perturbations in the pulmonary endothelial cells.

Influence of estrogen on glutathione levels and glutathione-metabolizing enzymes in uteri and R3230AC mammary tumors of rats

The glutathione content and the activities of several enzymes in its metabolism, glutathione reductase, glutathione peroxidase and γ-glutamyl transpeptidase, were assayed in uteri obtained from estrogen-treated rats and in R3230AC mammary adenocarcinomas obtained from ovariectomized, intact and estrogen-treated hosts. Normal mammary glands, obtained 10–12 days post-partum, were also examined for these parameters. A daily pharmacological dose of 0.4 μg of estradiol-17β induced a maximal increase in uterine weight and in reduced glutathione (GSH); higher doses of estrogen did not significantly increase either of these parameters. Levels of oxidized glutathione (GSSG) were comparable in both estrogen-treated and untreated rats. The time course of the estrogen-induced uterotrophic response was associated with increases in glutathione reductase, glutathione peroxidase and γ-glutamyl transpeptidase activities with the increased GSH level preceding the increase in uterine weight. Compared to neoplasms from intact or ovariectomized animals, tumors from estrogen-treated hosts exhibited significant decreases in levels of GSSG and GSH, as well as in glutathione reductase and glutathione peroxidase activities, but demonstrated a significant elevation of γ-glutamyl transpeptidase activity. Normal glands from lactating rats had decreased GSH levels, lower activities of glutathione reductase and glutathione peroxidase, but elevated γ-glutamyl transpeptidase activity versus tumors from intact rats. Tumors from estrogen-treated rats more closely resembled mammary glands during lactation. The divergent growth responses elicited by estrogen in the uterus and mammary tumor are correlated with the observed changes in GSH levels and enzymes involved in glutathione metabolism.