Severity of psychosis syndrome and change of metabolic abnormality in chronic schizophrenia patients: severe negative syndrome may be related to a distinct lipid pathophysiology.

BACKGROUND: Metabolic abnormality is common among schizophrenia patients. Some metabolic traits were found associated with subgroups of schizophrenia patients. OBJECTIVES: We examined a possible relationship between metabolic abnormality and psychosis profile in schizophrenia patients. METHOD: Three hundred and seventy-two chronic schizophrenia patients treated with antipsychotics for more than 2 years were assessed with the Positive and Negative Syndrome Scale. A set of metabolic traits was measured at scheduled checkpoints between October 2004 and September 2006. RESULTS: Multiple regressions adjusted for sex showed negative correlations between body mass index (BMI) and total score and all subscales; triglycerides (TG) was negatively correlated with total score and negative syndrome, while HDLC was positively correlated with negative syndrome. When sex interaction was concerned, total score was negatively correlated with BMI but not with others; negative syndrome was negatively correlated with BMI and positively with HDLC. No metabolic traits were correlated with positive syndrome or general psychopathology. CONCLUSIONS: Loss of body weight is a serious health problem in schizophrenia patients with severe psychosis syndrome, especially the negative syndrome. Schizophrenia patients with severe negative syndrome may have a distinct lipid pathophysiology in comparison with those who were less severe in the domain.

Ethanolic extracts of Antrodia cinnamomea mycelia fermented at varied times and scales have differential effects on hepatoma cells and normal primary hepatocytes.

Mycelia of Antrodia cinnamomea (AC), an edible fungus native to Taiwan, were produced by submerged fermentation with various fermentation times in 250 mL, 5 and 500 L fermentors and were evaluated for the effect of fermentation products on the viabilities of Hep3B and HepG2 hepatoma cells and normal primary rat hepatocytes. The results showed that the ethanolic extracts of AC mycelia (from 250 mL fermentation for 8 wk and 5 and 500 L fermentations for 4 wk) possessed high antihepatoma activity. The IC(50) of ethanolic extract of AC mycelia fermented for 8 wk in a 250 mL fermentor against Hep3B and HepG2 cells were 82.9 and 54.2 microg/mL, respectively. Furthermore, the IC(50) for Hep3B and HepG2, treated with ethanolic extract of AC mycelia fermented for 4 wk in the 5 L fermentor were 48.7 and 3.8 microg/mL, respectively. Those treated with ethanolic extract of AC mycelia fermented for 4 wk in the 500 L fermentor were 36.9 and 3.1 microg/mL, respectively. No adverse effects of all samples on normal primary rat hepatocytes were observed.

Inhibitory effect of CDA-II, a urinary preparation, on aflatoxin B(1)-induced oxidative stress and DNA damage in primary cultured rat hepatocytes.

The effects of CDA-II (cell differentiation agent II; a urinary preparation) on both aflatoxin B(1) (AFB(1))-induced cell injury and DNA damage were investigated using cultured rat hepatocytes. CDA-II was able to suppress both the lipid peroxidation and lactate dehydrogenase leakage induced by AFB(1). Glutathione (GSH) depletion by AFB(1) was replenished by CDA-II treatment. Under these experimental conditions, CDA-II enhanced the activity of GSH peroxidase, but not GSH S-transferase. By evaluation of unscheduled DNA synthesis, CDA-II reduced AFB(1)-induced DNA damage in hepatocyte cultures. These findings suggest that CDA-II can inhibit cytotoxicity of AFB(1) through enhancing the activity of GSH peroxidase and preventing GSH depletion.

Differential effects of allyl sulfides from garlic essential oil on cell cycle regulation in human liver tumor cells.

In this study, diallyl sulfide (DAS), diallyl disulfide (DADS) and diallyl trisulfide (DATS), which are major organosulfur compounds (OSCs) of garlic, were used as experimental materials to investigate their modulation effects on cell viability and cell cycle in human liver tumor cells (J5). According to the results of cell viability assay, 50 or 100 microM DATS significantly decreased the cell viability as compared with the control (P < 0.05) in dose and time dependent relations. Phenomena of cell number loss, shape deformation and lysis were observed after treatment with 100 microM DATS for 24 h. Cell cycle studies showed that J5 cells were significantly arrested in G2/M phase as the cells were treated with 100 microM DADS, 10, 50 or 100 microM DATS for 24 h (P < 0.05). DATS was more effective in arresting cells in G2/M phase than DADS, and the phenomena of arresting J5 cells in G2/M phase increased obviously in dose and time dependent relations. According to the Western blot analysis, DATS decreased cyclin-dependent kinase (Cdks)-Cdk7 (i.e. Cdc2 activate kinase) protein levels in J5 cells but increased cyclin B1 protein level. The modulation potency to cyclin B1 and Cdk7 expressions was in the order of DATS > DADS > DAS. The modulation potency to cyclin B1 and Cdk7 protein levels increased with increasing in DATS concentration and culture time. In conclusion, DATS might affect cell viability and cell morphological changes in J5 cells and lead cells to be arrested in G2/M phase via controlling the expression of cyclin B1 and Cdk7 in J5 cells, and the controlling action might relate to the sulfuric atom numbers in the structures of all these allyl sulfides.

Comparison of the metabolic effects of metformin and troglitazone on fructose-induced insulin resistance in male Sprague-Dawley rats.

BACKGROUND AND PURPOSE: Insulin resistance is a hallmark of the development of type 2 diabetes. Metformin and troglitazone are oral antidiabetic agents used to reduce insulin resistance. The aim of this study was to compare the metabolic effects of these two drugs in fructose-induced insulin-resistant rodents. METHODS: Male Sprague-Dawley rats were allocated to receive one of the following four treatments for 6 weeks: normal rat chow (control group, n = 7), high-fructose diet (fructose group, n = 7), high-fructose diet plus metformin (metformin group, n = 8), or high-fructose diet plus troglitazone (troglitazone group, n = 8). Systolic blood pressure (SBP), insulin, free fatty acid (FFA), and triglyceride concentrations were measured as parameters of insulin resistance. Leptin concentration was also measured in the four groups. RESULTS: The fructose group developed significantly elevated SBP, hyperinsulinemia, and hypertriglyceridemia without significant change in body weight or leptin concentration compared with the control group. The metformin group had significantly reduced body weight (397.9 +/- 40.9 vs 470.1 +/- 59.6 g, p < 0.05), insulin concentration (14.8 +/- 10.5 vs 48.4 +/- 15.2 microU/mL, p < 0.05), triglyceride concentration (75.3 +/- 65.5 vs 250.1 +/- 95.7 mg/dL, p < 0.05), and leptin concentration (3.1 +/- 1.5 vs 6.9 +/- 2.0 ng/mL, p < 0.05) without significant change in SBP (147.8 +/- 5.8 vs 152.4 +/- 13.0 mm Hg, p > 0.05) compared with the fructose group. The troglitazone group had significantly reduced SBP (137.8 +/- 9.2 vs 152.4 +/- 13.0 mm Hg, p < 0.05), insulin concentration (15.0 +/- 13.6 vs 48.4 +/- 15.2 microU/mL, p < 0.05), FFA concentration (38.9 +/- 22.7 vs 78.7 +/- 24.6 mg/dL, p < 0.05), triglyceride concentration (67.6 +/- 32.4 vs 250.1 +/- 95.7 mg/dL, p < 0.05), and leptin concentration (4.4 +/- 2.0 vs 6.9 +/- 2.0 ng/mL, p < 0.05) without significant change in body weight (452.5 +/- 32.8 vs 470.1 +/- 59.6 g, p > 0.05) compared with the fructose group. The metabolic effects of metformin and troglitazone on insulin, FFA, triglyceride, and leptin concentrations were not significantly different. However, metformin treatment resulted in significantly lower body weight (397.9 +/- 40.9 vs 452.5 +/- 32.8 g) and troglitazone treatment in significantly lower SBP (137.8 +/- 9.2 vs 147.8 +/- 5.8 mm Hg) compared to the fructose group, after adjusting for basal values (p < 0.05). CONCLUSIONS: Both metformin and troglitazone were comparably effective in reducing insulin resistance. Metformin treatment caused body weight reduction but was not effective in reducing SBP. Troglitazone treatment lowered SBP but did not reduce body weight.

Effect of diallyl sulfide and diallyl disulfide, the active principles of garlic, on the aflatoxin B(1)-induced DNA damage in primary rat hepatocytes.

The objective of this study was to investigate the effect of the active principles in garlic-- diallyl sulfide (DAS) and diallyl disulfide (DADS)--on aflatoxin B(1) (AFB(1))-induced DNA damage in primary rat hepatocytes. Primary rat hepatocytes, induced with DNA damage using 10 microM AFB(1) were used as an experimental model. According to the results of LDH leakage, 0.5 and 2 mM of DAS or 0.5 and 1 mM of DADS significantly increased the viability of hepatocytes compared with the AFB(1) controls after 4, 8 and 24 h treatment (P<0.05). According to the results of unscheduled DNA synthesis (UDS) test, 0.5 and 2 mM of DAS or 0.5 and 1 mM of DADS could significantly decrease the DNA damage induced by AFB(1) (P<0.05). Furthermore, 0.5 and 2 mM DAS or 0.5 and 1 mM DADS could increase the glutathione S-transferase (GST) and glutathione peroxidase (GPx) activities as compared with the AFB(1) controls after 24 h treatment (P<0.05). Results of immunoblot analysis of cytosolic GST isoenzyme indicate that the levels of GST isoform Ya, Yb2 and Yc were markedly increased after treatment with 0.5 and 2 mM DAS or 0.5 and 1 mM DADS compared with the AFB(1) control. These results indicate that 0.5 and 2 mM DAS or 0.5 and 1 mM DADS might protect hepatocytes from AFB(1)-induced DNA damage via increasing the activities of GST and GPx.

Dietary fat and garlic oil independently regulate hepatic cytochrome p(450) 2B1 and the placental form of glutathione S-transferase expression in rats.

The individual and combined effects of dietary fat and garlic oil on two drug-metabolizing enzymes, cytochrome P(450) 2B1 and the placental form of glutathione (GSH) S-transferase (PGST), in rat liver were examined in this study. Rats were fed a low corn oil, high corn oil or high fish oil diet and received various amount of garlic oil (0, 30, 80, 200 mg/kg body) orally three times per week for 6 wk. The fat energy in the low and high fat diets accounted for 11.6 and 45.7% of total energy, respectively. Final body weights did not differ among the three dietary fat groups and were not affected by garlic oil treatment. The fatty acid profile in hepatic phospholipids revealed higher eicosapentaenoic acid [20:5(n-3)] and docosahexaenoic acid [22:6(n-3)] levels in the fish oil-fed group than in the low and high corn oil-fed groups (P < 0.05). In contrast, the corn oil-fed groups had greater hepatic phospholipid arachidonic acid [20:4(n-6)] levels (P < 0.05). Both dietary fat and garlic oil significantly affected hepatic cytochrome 7-pentoxyresorufin O-dealkylase (PROD) activity and GST activity toward ethacrynic acid. Rats fed the high fish oil diet had 85 and 51% higher PROD activity compared with those fed the low or the high corn oil diet, respectively (P < 0.05). The GST activity in the high fish oil and the high corn oil groups was 33 and 18% higher than that in the low corn oil group (P < 0.05), respectively, and the GST activity in rats fed the high fish oil diet was higher than in those fed the high corn oil diet (P < 0.05). Garlic oil dose-dependently increased GST activity. No interaction between dietary fat and garlic oil on PROD or GST activity was noted. Northern and Western blot analysis revealed that dietary fish oil increased both cytochrome P(450) 2B1 and PGST mRNA and protein levels. Cytochrome P(450) 2B1 and PGST mRNA and protein levels were also dose-dependently increased by garlic oil treatment. The effects of garlic oil and dietary fat on P(450) 2B1 and PGST mRNA and protein expression were independent. These results indicate that dietary fat and garlic oil independently modulate P(450) 2B1 and PGST expression at transcriptional and/or post-transcriptional stages.

Effects of organosulfur compounds from garlic oil on the antioxidation system in rat liver and red blood cells.

The modulation of garlic oil (GO) and three allyl compounds, diallyl sulfide (DAS), diallyl disulfide (DADS) and diallyl trisulfide (DATS), on the antioxidation system in rat livers and red blood cells was examined. Rats were orally administered GO (200 mg/kg body weight), DAS (20, 80 mg/kg body weight), DADS (80 mg/kg body weight) or DATS (70 mg/kg body weight) three times a week for 6 weeks. Control rats received corn oil (2 ml/kg body weight) alone. GO, DADS and DATS treatment significantly increased the glutathione (GSH) content (48-84%) in red blood cells (P < 0.05). DATS displayed a greater enhancement than GO and DADS (P < 0.05). Hemolysis induced by tert-butyl hydroperoxide was not suppressed by GO or allyl compound treatment although higher GSH content was evident. Hepatic GSH was not influenced by garlic components. In rat livers, DADS and DATS significantly increased the activity of GSH reductase (46 and 54%, respectively) and of GSH S-transferase (GST) (63 and 103%, respectively), but decreased the GSH peroxidase activity (27 and 28%, respectively). In contrast, GSH reductase and GST activities in the DAS group, either 20 or 80 mg/kg body weight, were similar to the control group. A decrease of GSH peroxidase activity was observed in rats dosed with 80 mg/kg body weight (P < 0.05). An increase in GST activity and a decrease in GSH peroxidase activities were also noted in GO-treated rats (P < 0.05). In red blood cells, three GSH-related antioxidant enzyme activities were not affected by garlic oil and its organosulfur components. Immunoblot assay showed that, accompanying the increase in hepatic GST activity, GO, DADS, DAS (80 mg/kg body weight) and DATS increased the expression of GST Ya, Yb1 and Yc proteins. Results indicate that GO and three allyl compounds play a differential role in modulation of the GSH-related antioxidant system in rat livers and red blood cells.

Effect of garlic active principle, diallyl disulfide, on cell viability, lipid peroxidation, glutathione concentration and its related enzyme activities in primary rat hepatocytes.

This study investigated the effects of various concentrations and incubation time intervals of diallyl disulfide (DADS), the active principle of garlic, on: 1. cell viability, 2. lipid peroxidation, and 3. glutathione (GSH) concentration and its related enzyme activities of rat hepatocytes. According to the results of LDH leakage and microscopic examination, 0.5 and 1 mM DADS did not significantly affect the viability of hepatocytes. However, significant decrease in cell viability according to increased LDH leakage and significant changes in morphology of hepatocytes were observed at 2 mM DADS (P < 0.05). Lipid peroxidation was also detected when the hepatocytes were treated with 2 mM DADS. At 0.5 mM DADS, a higher GSH content was found in the hepatocytes although not at a statistically significant level. 0.5 and 1 mM DADS has little effect on the activities of glutathione-S-transferase (GST) and glutathione peroxidase (GPx); however a significant decrease in GST, GPx and glutathione reductase (GRd) activities was observed at 2 mM DADS. Once the media of 2 mM DADS was replaced with fresh medium at 24 hr treatment, the activities of GST, GRd and GPx were recovered, although they were still lower than the control values.

Metabolites of diallyl disulfide and diallyl sulfide in primary rat hepatocytes.

The objectives of this study were to analyse and identify the metabolites of diallyl disulfide (DADS) and diallyl sulfide (DAS) in primary rat hepatocytes prepared by collagenase perfusion. According to the results, allyl mercaptan (AM) and allyl methyl sulfide (AMS) were the metabolites of DADS. The highest amount of AMS (0.93 +/- 0.08 microg/ml at 90 min) was much less than that of AM (46.2 +/- 6.6 microg/ml at 60 min). Combined with the Purge and Trap using a gas chromatography-mass spectrometry (GC-MS) system, it is very useful to detect the trace amounts of metabolites in primary rat hepatocytes. Results also showed that AMS was a metabolite of DAS. The highest amount of AMS in the extracellular fluid of hepatocytes was 0.63 +/- 0.16 microg/ml at 30 min of incubation.

Effects of garlic oil and its organosulfur compounds on the activities of hepatic drug-metabolizing and antioxidant enzymes in rats fed high- and low-fat diets.

We examined the effects of garlic oil (GO) and two of its organosulfur compounds, diallyl sulfide (DAS) and diallyl disulfide (DADS), on the drug-metabolizing and antioxidant systems in rats and sought to determine whether these effects are associated with dietary fat. Rats were fed a high-fat diet and received GO or DADS (200 mg/kg body wt) or DAS (100 mg/kg) orally three times a week for seven weeks. Control animals received corn oil alone. Another group of rats was fed a low-fat diet, with or without GO. GO and DADS significantly reduced the body weight gain of rats (p < 0.05). GO, however, dramatically increased the spleen weight and spleen weight-to-body weight ratio (p < 0.05). DAS increased glutathione S-transferase (GST) and 7-pentoxyresorufin O-dealkylase activities, whereas DADS increased only GST activity (p < 0.05). Immunoblot assay showed GO-, DAS-, and DADS-enhanced expression of the placental form of GST and cytochrome P-450 IIBI but suppressed cytochrome P-450 IIEI expression. Hepatic antioxidant enzyme activities were also modulated by these garlic components. GO and DADS inhibited glutathione peroxidase activity (p < 0.05), and DADS and DAS enhanced glutathione reductase activity (p < 0.05). Only GO enhanced the superoxide dismutase activity (p < 0.05). All these garlic components increased glutathione levels in red blood cells (p < 0.05) but did not influence hepatic glutathione levels. Although the amount of fat in the diet modulated drug-metabolizing and antioxidant functions, no interactions between GO and dietary fat were observed. These results indicate that GO and its allyl sulfide components, as well as dietary lipid, modulate drug-metabolizing and antioxidant enzyme activities. The action of GO appears to be independent of dietary lipid content.

Methionine and cysteine affect glutathione level, glutathione-related enzyme activities and the expression of glutathione S-transferase isozymes in rat hepatocytes.

Methionine and cysteine are constituents of glutathione. To understand the effects of these two sulfur amino acids on the glutathione (GSH)-dependent detoxification defense system, intracellular GSH and GSH-related enzyme activities, including GSH peroxidase, GSH reductase, GSH S-transferase (GST) and gamma-glutamylcysteine synthetase, were determined. In addition, the expression of three GST isozymes and carbonic anhydrase III (CA III) was examined. Hepatocytes isolated from male Sprague-Dawley rats were cultured with 0.1, 0.3, 0.5 or 1.0 mmol/L each of L-methionine and L-cysteine, for up to 7 d. Cells incubated with 0.5 or 1.0 mmol/L methionine and cysteine had increased intracellular GSH. A twofold increase was observed on d 6 compared with freshly isolated hepatocytes (P < 0.05). However, intracellular GSH was lower in cells treated with 0.3 or 0.1 mmol/L each of methionine and cysteine than in cells tested with 0.5 or 1.0 mmol/L. Although the GSH level differed significantly between cells cultured with 0.3 or 1.0 mmol/L of methionine and cysteine, GSH-related enzymes did not differ at these two concentrations. The activity generally remained constant for the first 24 h, then increased up to d 4. Immunodetection analysis revealed no difference in the level of CA III and GST isoforms, Ya, Yb and Yp, with amino acids each at a concentration of at least 0.3 mmol/L. Yp expression steadily increased up to d 7. Most proteins decreased rapidly after 48 h when cultured with 0.1 mmol/L of methionine and cysteine; however, the Yp level increased up to d 6. In conclusion, results indicate that a twofold increase of intracellular GSH is reached by adding methionine and cysteine at a concentration >0.5 mmol/L to the culture medium. The concentrations of methionine and cysteine for maintaining hepatic GSH are higher than for GSH-related enzyme activity and for GST isoform expression.

Corn oil enhances gamma-glutamyl transpeptidase-positive foci development in the presence of phenobarbital during hepatocarcinogenesis in rats.

We investigated the effects of type of dietary fat and phenobarbital on gamma-glutamyl transpeptidase-positive foci development. Four groups of six female Sprague-Dawley rats were initiated with diethylnitrosamine (15 mg/kg) at 24 hours of age. After weaning, they were fed nutritionally complete semipurified diets containing 15% corn oil or 5% corn oil + 10% fish oil and supplemented with 5,000 ppm vitamin E with or without phenobarbital (500 ppm) for three months. Dietary fish oil significantly increased hepatic phospholipid eicosapentaenoate and docosahexaenoate concentrations and decreased arachidonate concentration compared with 15% corn oil (p < 0.05). Corn oil (15%) significantly increased hepatic prostaglandin F2 alpha concentration compared with 10% fish oil (p < 0.05). Phenobarbital significantly stimulated glutathione S-transferase activity in both dietary fat groups (p < 0.05). In the absence of phenobarbital, type of dietary fat showed no effect on hepatic gamma-glutamyl transpeptidase-positive foci development. However, in the presence of phenobarbital, 15% corn oil significantly enhanced gamma-glutamyl transpeptidase-positive foci development compared with 10% fish oil (p < 0.05). Phenobarbital showed a strong tumor-promoting action in both dietary groups. In conclusion, there was an interaction between type of dietary fat and phenobarbital on gamma-glutamyl transpeptidase-positive foci development during hepatocarcinogenesis in rats.

Amount and type of dietary lipid modulate rat hepatic cytochrome P-450 activity.

The influence of the amount and type of dietary lipid on rat hepatic cytochrome P-450 activities in the presence and absence of inducer administration was investigated. Weanling male Sprague-Dawley rats were fed fat-free or 20% beef tallow, olive oil, corn oil, linseed oil, or menhaden oil diets in combination with one of the following three treatments: no inducer, intraperitoneal injection of phenobarbital (75 mg/kg body wt) for three consecutive days before they were killed, or intragastric administration of acetone (5 ml/kg) one day before they were killed. Twenty percent linseed oil and menhaden oil diets induced the highest level of activity among the different fat types in the presence of phenobarbital and acetone. Cytochrome P-450IIB1 activity was induced to a significantly greater extent by acetone administration in conjunction with the 20% menhaden oil diet than in conjunction with the other dietary oils (p < 0.05). In the presence of acetone, 20% beef tallow, 20% linseed oil, and 20% menhaden oil diets significantly induced cytochrome P-450IIE1 activity compared with the fat-free diet (p < 0.05). In conclusion, cytochrome P-450IIB1 and P-450IIE1 activities in rats were significantly increased by specific inducers, and dietary lipid was necessary for this effect. Diets supplemented with linseed and menhaden oils were most effective in inducing this activity.

Effect of the active principle of garlic--diallyl sulfide--on cell viability, detoxification capability and the antioxidation system of primary rat hepatocytes.

The objectives of this study were to investigate the effects of various concentrations and incubation time intervals of diallyl sulfide (DAS), an active principle of garlic, on cell viability, and glutathione (GSH) concentration and its related enzymes activities in rat hepatocytes. According to the results of lactate dehydrogenase (LDH) leakage and microscopic examination, 0.5 or 1 mM DAS treatment did not have any adverse effects on the viability of hepatocytes. Intracellular GSH contents of cells treated with 0.5 and 1 mM DAS (58.6 and 66.4 nmol GSH/mg protein, respectively) were higher than in the controls (54.2 nmol GSH/mg protein), around 8-23%, at 24 hr of incubation; a significant difference (P < 0.05) was observed for 1 mM DAS treatment at 48 hr. This phenomenon is beneficial to the detoxification and antioxidation capabilities of hepatocytes. Further, when the hepatocytes were treated with 0.5 or 1 mM DAS, the activities of glutathione S-transferase (GST), glutathione peroxidase (GPx) and glutathione reductase (GRd) were almost the same as those of the controls. On the other hand, treatment with 5 mM DAS was associated with a significant decrease (P < 0.05) in cell viability, namely in increased LDH leakage (50% at 24-hr treatment), significant changes in the morphology of the hepatocytes, low intracellular GSH level (45% lower than in the controls at 24-hr treatment), and low activities of GST, GPx and Grd.

Glutathione and glutathione-related enzyme activities of male and female rat hepatocytes under various culture conditions.

The effect of culture medium on glutathione (GSH) dependent detoxification defence system of primary cultured hepatocyte from either male or female rats was studied. Intracellular reduced (GSH) and oxidized glutathione (GSSG), and six GSH-related enzyme activities, including GSH peroxidase (GSH Px), GSH reductase (GSH Rd), cytosolic GSH S-transferase (cGST), microsomal GSH S-transferase (mGST), gamma-glutamyl transpeptidase (GTP), and gamma-glutamylcysteine synthetase (GCS), were investigated during a 6-day culture. Media free of fetal bovine serum (FBS) and with 2.5 or 10% FBS were used. Whatever the medium, there was an initial decrease of intracellular GSH and GSSG, a threefold increase of GSH at day 3 and fourfold increase of GSSG at day 4, later decreasing to their original level at day 6. The activities of all six GSH-related enzymes of male and female hepatocytes remained relatively stable during the first 72h, then gradually decreased to 50-80% of initial activities. With the exception of cGST, time-course profiles of other enzyme activities were not significantly different among various media. In both sexes, higher cGST activity was maintained for cells cultured in the presence of FBS. Results of immunoblotting analysis of cytosolic GST isozymes indicate that the placental form of GST (Yp) was markedly increased after plating and the extent of increase of Yp was higher in the presence of FBS. Despite the culture medium, the level of GST isoform Ya was maintained steadily for 6 days, however, Yb was maintained during the first 3 days and then decreased. In terms of the gender difference, GSH Px and GTP activities of hepatocytes from females were significantly greater than of males over the entire culture period. Results indicate that FBS seems not to be absolutely essential in maintaining GSH level and most of the GSH-related enzyme activities in rat hepatocytes. Furthermore, GSH levels and GSH-related enzyme activities of hepatocytes from female rats were similar to those from male rats.