The effect of selenium on the tissue sulfhydryl group content and lipid peroxide-destorying enzyme system in the liver, kidney and testis of rat treated with mercury was investigated. The male rats were injected s.c. with HgCl2 (10 micromoles/kg BW) and orally received Na2SeO3 (13 micromoles/kg BW) simultaneously. After 3 days, liver, kidney and testis were removed and analyzed. Mercury decreased the total sulfhydryl group content in the kidney by 25% and the total glutathione content in the kidney and testis by 50% and 36%, respectively, with no changes in other tissues. There was 12% increase in the total sulfhydryl group but not in the total glutathione content in kidney by a simul-taneous treatment of Se and Hg. Glutathione peroxidase (GSH-Px) activities were decreased by 63% in the liver and 69% in the kidney, and glutathione reductase (GSH-Rd) activity was increased in the tests by 16% by the Hg treatment with no changes in Other tissues. Hg had no effect upon glutathione-S-transferase activities in all organs examined. Simultaneous Se treatment increased GSH-Rd activity in the kidney by 23% and GSH-Px activities in liver and kidney by 24% and 21%, respectively, compared to the Hg-treated group. These data indicate that the alleviation of Hg toxicity by Se treatment is well correlated with the protein sulfhydryl group content and GSH-Px activity.
Animal ; Glutathione/metabolism* ; Glutathione Peroxidase/analysis ; Glutathione Reductase/analysis ; Male ; Mercury/toxicity* ; Rats ; Selenium/pharmacology* ; Sulfhydryl Compounds/analysis*

To identify the inhibitor of glutathione S-transferase (GST), a high-throughput screening method was established in a 384-well microplate with total 35 microL volume, and the absorbance at 340 nm is detected. The concentrations of substrates, CDNB and GST were determined by chromatometry. The optimal enzyme kinetics reaction time and temperature are 2 h and 30 degrees C , respectively. The established model was evaluated by NaOCl, a known GST inhibitor, and the parameter Z' was 0.77, which showed a high feasibility and stability of the assay. A total of 31,098 compounds were screened, of which 4 compounds were shown to inhibit GST activity, high inhibiting activity for their IC50 of GST inhibition was 3.94, 4.05, 74.85, and 77.41 mg x L(-1), separately. The results indicated that the colorimetric method by using CDNB and GSH as substrate is stable, sensitive, reproducible and also suitable for high throughput screening.
Dinitrochlorobenzene ; chemistry ; Drug Evaluation, Preclinical ; methods ; Enzyme Inhibitors ; analysis ; Glutathione ; chemistry ; Glutathione Transferase ; antagonists & inhibitors ; Substrate Specificity

An experiment was conducted in order to investigate the effect of p-dimethylaminoazobenzene (DAB) and 2(3)-tert-butyl-4-hydroxyanisole (BHA) on the lipid peroxidation and peroxide-destroying enzyme system in the rat liver. Dietary supplementation of DAB (0.06%) for three weeks caused the elevation of glutathione-S-transferase activity by 60% and glutathione reductase by 50%, but it decreased glutathione peroxidase and catalase activities significantly. Dietary supplementation of BHA (0.75%) also increased glutatione-S-transferase activity in the liver by 2 folds, and it counteracts DAB effect on the glutathione peroxidase and catalase activities. There was a marked increase in malon-dialdehyde content in the postnuclear fraction of liver by the treatment of DAB, but the addition of BHA lowered the malondialdehyde content to almost the control level. The protective effect of BHA on the lipid peroxidation induced by DAB administration at the enzyme level seems to be due to the induction of glutathione-S-transferase and the protection of glutathione peroxidase and catalase activities from being lowered by DAB administration.
Animal ; Anisoles/pharmacology* ; Butylated Hydroxyanisole/pharmacology* ; Glutathione Peroxidase/analysis* ; Glutathione Reductase/analysis* ; Glutathione Transferase/analysis* ; Lipid Peroxides/metabolism* ; Liver/drug effects* ; Liver/metabolism ; Male ; Peroxidases/analysis* ; Rats ; p-Dimethylaminoazobenzene/pharmacology*

OBJECTIVESTo identify hepatic progenitor cells (HPCs) in patients with severe hepatitis (SH) by detecting their markers and investigate the features of their distribution and location.

METHODSLiver tissues taken from 59 SH patients were tested for the receptor of stem cell factor (c-kit), pi-class glutathione S-transferase (GST-pi), cluster of differentiation 34 (CD34), cytokeratin 19 (CK19), cytokeratin 18 (CK18) and alpha fetoprotein (AFP) by immunohistochemistry (IHC). Meanwhile, 58 patients with acute or chronic hepatitis were also detected to act as controls.

RESULTSHepatic progenitor cells could be seen in SH patients. Most of them existed as ductular cells that had been called "typical ductular proliferation (ADP)" or "typical ductular reaction" in previous research. These ductular cells were mainly located at the portal areas, fibro septa, periportal parenchyma and the border of the pseudolobuli and inflammatory foci. Further, c-kit, GST-pi, CK19 and CK18, but not CD34 and AFP could be detected in these cells. Another kind of HPC was the small hepatocyte-like cell (SHLC), which could express c-kit, GST-pi, and CK18, but not CK19, CD34 and AFP. The semi-quantitative analysis showed that the scope of ADP in SH patients was significantly larger than that in acute and chronic hepatitis patients (chi2= 63.62, P<0.05), and the scope of ADP in subacute severe hepatitis and chronic severe hepatitis patients was also significantly larger than that in acute severe hepatitis patients.

CONCLUSIONIn the course of regeneration of viral hepatitis, different types of pathology have different features. In acute and chronic hepatitis (G1-2), the regeneration is mainly owing to the proliferation of mature hepatocytes, and in chronic hepatitis (G3-4), there is the participation of HPCs, although they are limited. In severe hepatitis, however, since the replicative capacity of normal hepatocytes is impaired or prohibited, liver regenerates and restores mainly by the means of hepatic stem cells activation and proliferation. But the hepatic stem cells don't differentiate into their mature functional compartments directly at all. There are several intermediary or transition populations. In human severe hepatitis, they are mainly ductular cells, and parts of them are small hepatocyte-like cells.

Antigens, CD34 ; analysis ; Cell Division ; Female ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Hepatitis ; pathology ; Hepatocytes ; pathology ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Keratins ; analysis ; Male ; Proto-Oncogene Proteins c-kit ; analysis ; Stem Cells ; pathology ; alpha-Fetoproteins ; analysis

To develop a method for the detection of surface-confined peptides containing cysteine residues or oligodeoxynucleotides (ODNs) whose 3' ends modified with thiol groups, and a thiol-specific fluorescent cross-linker, N-(9-acridinyl) maleimide (NAM) was used. The peptides studied herein include both the oxidized and reduced forms of glutathione, and a hexapeptide (FT). Peptides are first attached onto the activated 11-mercaptoundecanoic acid (MUA)-terminated alkanethiol self-assembled monolayers (SAMs) and then derivatized with NAM. The cysteine residues was determined by using electrochemical desorption and fluorescence detection. GSH concentration as low as 40 pmol x L(-1) can be measured. The fluorescence intensity in the case of FT is about 3 times as high as that for GSH, which is consistent with the molar ratio of cysteine residues in these two molecules. The analytical performance of gene analysis was also evaluated through the analyses of a complementary target and targets with varying numbers of mismatching bases. The method described here is simple, sensitive, reproducible, and does not require sophisticated analytical instrumentation and separation procedures.
Biosensing Techniques ; methods ; Cysteine ; analysis ; Electrochemistry ; methods ; Fluorescence ; Glutathione ; analysis ; chemistry ; Maleimides ; chemistry ; Oligodeoxyribonucleotides ; analysis ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo explore the correlation among some biochemical indexes in the fluoride workers.

METHODSThe activities of superoxide dismutase(SOD), glutathione peroxidase (GSH-Px), catalase (CAT), alkaline phosphatase (AKP) and the level of calcitonin (CT), parathyroid hormone (PTH), IgG, IgA, IgM, Cu2+, Zn2+, Ca2+, Mg2+ and Se2+, F- in serum and in urine were measured in fifty male fluoride workers and fifty controls.

RESULTSThe levels of F-, CT, PTH, AKP and GSH-Px in serum and F- in urine in exposed group were significantly different from that in control group. Correlation analysis indicated that F- in urine and CAT(r = 0.3133, P < 0.05), CT and PTH(r = 0.5173, P < 0.01), Se2+ and CAT(r = 0.4354, P < 0.05) were positively correlated. There were significantly negative correlation between F- in serum and GSH-Px (r = -0.5202, P < 0.01) and positive correlation among Cu2+, Zn2+, Ca2+ and Mg2+ in serum.

CONCLUSION(1) Excess of fluoride may affect secretion of calcium adjusting hormone (CT, PTH); (2) Changes of AKP and GSH-Px may be regarded as health monitoring indexes; (3) The correlation of biochemical indexes plays an important role in studying the mechanism and the early prevention and treatment of industrial fluorosis.

Alkaline Phosphatase ; analysis ; Calcitonin ; analysis ; Catalase ; analysis ; Environmental Monitoring ; Fluorides ; toxicity ; Glutathione Peroxidase ; analysis ; Humans ; Male ; Occupational Exposure ; Parathyroid Hormone ; analysis ; Superoxide Dismutase ; analysis

This study was undertaken to investigate the antioxidant/oxidant status in recurrent miscarriage patients. Antioxidants including glutathione peroxidase (GPx), catalase (CAT), glutathione reductase (GR), reduced glutathione (GSH) and selenium (Se), as well as the oxidants hydrogen peroxide (H2O2), oxidised glutathione (GSSG) and lipid peroxidation were assayed in plasma, whole blood and placental tissue of non-pregnant women (NP), healthy pregnant women (HP), and recurrent miscarriage (RM) patients. Results indicated that all antioxidant activities and levels in plasma and whole blood of HP women were consistently moderately lower, and much more significantly lower in RM patients when both were compared to those seen in NP women (P<0.05 and P<0.001, respectively). Furthermore, whereas plasma antioxidant activities and levels were significantly lower in RM patients, those of whole blood and placental tissue were much more significantly lower when compared with HP women (P<0.001). Concurrent with these findings there were consistent increases of equal statistical significance and magnitude in the levels of all investigated oxidants assayed in all samples when compared in between subjects of the study as indicated above. Data thus illustrated a distinct shift in favor of oxidative reactions and reactive oxygen species (ROS) generation, and very significant decreases in the GSH/GSSG ratios in whole blood and placental tissue of RM patients when compared to HP and NP women (P<0.001). The above noted oxidative stress could have been a major causative factor of recurrent miscarriage.
Abortion, Habitual/*blood/*epidemiology ; Adult ; Antioxidants/analysis ; Biomarkers/*blood ; Catalase/blood ; Female ; Glutathione/blood ; Glutathione Peroxidase/blood ; Glutathione Reductase/blood ; Humans ; Hydrogen Peroxide/analysis ; Lipid Peroxidation ; *Oxidative Stress ; Placenta/metabolism ; Pregnancy ; Reactive Oxygen Species/metabolism ; Saudi Arabia/epidemiology ; Selenium/blood

OBJECTIVETo study the relationship between glutathione S-transferase M1 (GSTM1) genotypes and lipid peroxidation of asbestos workers.

METHODS94 asbestos workers and 51 controls were selected as subjects. The general information, occupational history and individual habits were collected by questionnaires in all participants. The venous blood was sampled and the plasma was separated for the detection of malondialdehyde (MDA) level and lymphocytes for DNA isolation and GSTM1 genotyping.

RESULTSMDA level was significantly higher in asbestos workers [(0.283 +/- 0.054) nmol/L] than that in controls [(0.163 +/- 0.053) nmol/L, P < 0.01], however, neither duration of exposure nor accumulated asbestos exposure dose was related to MDA levels; MDA levels in control workers with GSTM1 +/- genotype [(0.190 +/- 0.034) nmol/L] were significantly higher than that in control workers with GSTM1 +/+ genotype [(0.138 +/- 0.055) nmol/L, P < 0.01]. Among asbestos workers, the same trend could be found, but the differences was not significant(P > 0.05). When the workers were stratified by duration of exposure or accumulated asbestos exposure dose, MDA levels in individuals with GSTM1 -/- genotype were also higher than those with GSTM1 +/+ genotype, but the differences were also not significant(P > 0.05).

CONCLUSIONBoth exposure to asbestos and deficiency of GSTM1 genotype were related to lipid peroxidation in workers, but the role of the former may be more important than that of the latter.

Asbestos ; adverse effects ; Genotype ; Glutathione Transferase ; genetics ; Humans ; Lipid Peroxidation ; Malondialdehyde ; analysis ; Occupational Exposure

OBJECTIVETo investigate the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-hydroxypyrene (1-OHP) excretions in workers under different exposure levels.

METHODSFour hundred and forty-seven occupationally exposed workers from two coking plants and 220 control workers from a wire rod plant were genotyped to analyze the modification of GSTM1, GSTT1 and GSTP1 gene polymorphisms on urinary 1-OHP excretions.

RESULTSThe urinary 1-OHP concentration in exposed group was much higher than that in control group (4.61 vs 0.34 µmol/mol Cr, P < 0.05). Occupational exposure levels and cigarette smoking were of the dominating factors affecting 1-OHP excretions in urine. After controlling potential confounders, decreased excretion of urinary 1-OHP was associated with GSTP1 I105V AG + GG genotype in coke oven workers (single-gene model, P = 0.012; multi-gene model, P = 0.011) and with GSTT1 null type in the analysis including all subjects (P = 0.055 in both single-gene and multi-gene models). GSTT1 and GSTP1 were interacted on the urinary concentrations of 1-OHP.

CONCLUSIONUrinary 1-OHP concentrations can be modified by GSTM1, GSTT1 and GSTP1 gene polymorphisms, indicating that these genes are involved in the metabolism of polycyclic aromatic hydrocarbons.

Adolescent ; Adult ; Control Groups ; Genotype ; Glutathione S-Transferase pi ; genetics ; Glutathione Transferase ; genetics ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Polymorphism, Single Nucleotide ; Pyrenes ; analysis ; Urinalysis ; Young Adult

OBJECTIVETo investigate the effect of Coriolus versicolor polysaccharide B (CVP-B) on increased membrane glycosaminoglycans (GAG) expression and intracellular glutathione (GSH) of RAW264.7 macrophages exposed to angiotensin II (Ang II).

METHODSThe plasma membrane of RAW264.7 macrophages exposed to Ang II treatment was isolated by ultracentrifugation, and the membrane GAG expression was analyzed using 1, 9-dimethylmethylene blue (DMMB) spectrophotometric assay for sulfated GAG. The intracellular reduced GSH was determined using fluorophotometry.

RESULTSThe GAG content in the macrophage membranes increased by up to 54% following cell exposure to 1.0 micromol/L Ang II, whereas in presence of 1.0 micromol;/L Ang II, CVP-B at 1, 10, and 50 microg/ml decreased the GAG content by 13%, 43% (P<0.01), and 52% (P<0.01), respectively. The macrophage GSH activity decreased by 69% following incubation with 1.0 micromol;/L Ang II for 24 h, and CVP-B treatment at 1, 10, and 50 microg/ml in presence of 1.0 micromol;/L Ang II resulted in significant increment of GSH activity by 31%(P<0.05), 104% (P<0.01), and 168% (P<0.01), respectively.

CONCLUSIONThese data provide the first evidence that CVP-B inhibits elevated GAG expression in RAW264.7 macrophage membrane induced by Ang II.

Agaricales ; chemistry ; Angiotensin II ; pharmacology ; Animals ; Cell Line ; Cell Membrane ; metabolism ; Glutathione ; analysis ; Glycosaminoglycans ; analysis ; Macrophages ; metabolism ; Mice ; Polysaccharides ; pharmacology