共查询到20条相似文献，搜索用时 15 毫秒
1.
The glutathione S-transferases (GST) are a major group of conjugative enzymes involved in the detoxification of electrophilic compounds and products of oxidative stress. We have previously described the kinetics of hepatic GST conjugation in largemouth bass using a variety of synthetic GST reference substrates. In the present study, we investigated the ability of largemouth bass hepatic GSTs to conjugate 4-hydroxynon-2-enal (4HNE), a mutagenic and cytotoxic alpha-beta-unsaturated aldehyde produced during oxidative injury. Hepatic cytosolic fractions from largemouth bass rapidly catalyzed GSH-dependent 4HNE conjugation, with the rate of GST-4HNE conjugation in bass liver exceeding those of several other mammalian and aquatic species. No apparent sex-related differences in GST-4HNE activity were observed among adult bass. SDS-PAGE and Western blotting analysis of GSH affinity-purified bass liver cytosolic GST revealed the presence of two major GST subunits of approximately 30 and 27 KDa that exhibited slight cross-reactivity when probed with a rat alpha class GST antibody, but not to rat mu, pi or theta class GST. The rapid conjugation of 4HNE by hepatic GST suggests an important role for GSTs in protecting against peroxidation of polyunsaturated fatty acids in bass liver. 相似文献
2.
A cDNA clone for glutathione S-transferaseA (GSTA) from plaice (Pleuronectes platessa) was expressed in Eschericia coli (E. coli) and purified to homogeneity by S-hexylglutathione affinity chromatography. When compared to literature values for a variety of purified mammalian GSTs, the heterologously expressed purified plaice enzyme had moderate activity towards the model substrate 1,2-chloro-2,4-dinitrobenzene (CDNB) and exhibited a Km of 2.5 ± 2 mM and Vmax of 30.9 ± 2.3 μmol min−1 mg−1. It had little or no activity towards several other model GST substrates including 1,2-dinitrochloro-4-benzene (DCNB), ethacrynic acid (EA), and p-nitrobenzylchloride (NBC). However plaice GSTA was a relatively efficient catalyst for the conjugation of a series of alk-2-enals and alk-2,4-dienals and also 4-hydroxynonenal. The highest activity observed with this series of substrates was with trans-non-2-enal with a Km of 17.9 ± 2.2 μM and a Vmax of 3.01 ± 0.57 μmol min−1 mg−1. These unsaturated alkenals have been identified in cells and cell extracts as highly toxic products arising from peroxidation of unsaturated fatty acids particularly during periods of oxidative stress. Fish are relatively rich in polyunsaturated fatty acids and thus GSTA mediated conjugation may be an important mechanism for detoxifying peroxidised lipid breakdown products. 相似文献
3.
Almli B Egaas E Christiansen A Eklo OM Lode O Källqvist T 《Marine environmental research》2002,54(3-5):237-240
In order to evaluate the gill glutathione S-transferase (GST) activity as a biomarker of effect of fungicide exposure in juvenile brown trout (Salmo trutta), the fungicides propiconazole [(R,S)-1-[2-(2,4-diclophenyl)-4-propyl-1,3-dioolan-2-ylmetyl]-1H-1,2,4-triazole] and fenpropimorph [(+/-)-cis-4-[3-(4-tert-butylphenyl)-2-metyl propyl]-2,6 dimetylmorfolinc] were administrated in the water separately and together in a static system (80 microg/l for each pesticide) for 5 days. The combined fungicides gave a significant decrease in gill GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB), whilst hepatic GST-activity was not significantly changed. Furthermore, continuous exposure to 540 ug/l thiabendazole[2-(thiazol-4'-yl)benzimidazole] in a flow-through system for 4 days significantly increased the gill glutathione S-transferase (GST) activity towards CDNB, whilst hepatic GST and cytochrome P450 (CYP 1A) activities were not increased by the treatment. 相似文献
4.
Differential expression of alpha-like glutathione S-transferase (GST) isoforms in catfish intestine.
Previous studies suggested that dietary composition affected glutathione S-transferase (GST) activity in catfish intestine, and this activity varied along the intestine. In this study, catfish were fed a semi-purified diet or a commercial chow for at least 2 weeks. GST activity, percent protein cross-reacting with anti-catfish GST pi antibody, and immuno-cross-reactivity with antibodies specific for human alpha, mu, pi and theta class GSTs were determined in cytosol prepared from sections of proximal, medial, and distal intestine. The bulk of GST activity with 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid, and the percent protein cross-reacting with anti-catfish GST pi were in the more proximal segments and dropped off distally in the two diet groups. The percent of cross-reacting GST protein in the proximal section of fish fed on commercial chow was significantly higher (4.3 +/- 1.7%) than in fish fed purified diet (2.3 +/- 0.2%). Further Western blot analysis revealed a differential expression of GST isoforms only in the distal segment of fish fed commercial chow that recognized human anti-alpha GST antibody. Distal intestinal segments of catfish exposed to 3,3',4,4'-tetrachlorobiphenyl (TCB) and beta-naphthoflavone (BNF) also revealed expression of distinct alpha-like GST isoforms. Results strongly suggest the distal segment as a site for potential biomarkers for polycyclic aromatic hydrocarbon (PAH)- and co-planar polychlorinated biphenyl (PCB)-type contaminants. 相似文献
5.
B. Almli E. Egaas A. Christiansen O. M. Eklo O. Lode T. Kllqvist 《Marine environmental research》2002,54(3-5)
In order to evaluate the gill glutathione S-transferase (GST) activity as a biomarker of effect of fungicide exposure in juvenile brown trout (Salmo trutta), the fungicides propiconazole {(R,S)-1-[2-(2,4-diclophenyl)-4-propyl-1,3-dioolan-2-ylmetyl]-1H-1,2,4-triazole} and fenpropimorph {(±)-cis-4-[3-(4-tert-butylphenyl)-2-metyl propyl]-2,6 dimetylmorfolinc} were administrated in the water separately and together in a static system (80 μg/l for each pesticide) for 5 days. The combined fungicides gave a significant decrease in gill GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB), whilst hepatic GST-activity was not significantly changed. Furthermore, continuous exposure to 540 ug/l thiabendazole{ 2-(thiazol-4′-yl)benzimidazole} in a flow-through system for 4 days significantly increased the gill glutathione S-transferase (GST) activity towards CDNB, whilst hepatic GST and cytochrome P450 (CYP 1A) activities were not increased by the treatment. 相似文献
6.
We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to beta-naphthoflavone (beta-NF, 66 mg/kg, i.p.) elicited a 7-9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to beta-NF, and generally tracked the minimal changes observed in GST-CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by beta-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to beta-NF. 相似文献
7.
We are investigating the effects of in vivo exposure of prototypical enzyme inducing agents on hepatic biotransformation enzyme expression in largemouth bass (Micropterus salmoides), a predatory game fish found throughout the United States and Canada. The current study targeted those genes involved in biotransformation and oxidative stress that may be regulated by Ah-receptor-dependent pathways. Exposure of bass to β-naphthoflavone (β-NF, 66 mg/kg, i.p.) elicited a 7–9-fold increase in hepatic microsomal cytochrome P4501A-dependent ethoxyresorufin O-deethylase (EROD) activities, but did not affect cytosolic GST catalytic activities toward 1-chloro-2,4-dinitrobenzene (CDNB) or 5-androstene-3,17-dione (ADI). Glutathione S-transferase A (GST-A) mRNA expression exhibited a transient, but non-significant increase following exposure to β-NF, and generally tracked the minimal changes observed in GST–CDNB activities. Expression of the mRNA encoding glutamate-cysteine ligase catalytic subunit (GCLC), the rate-limiting enzyme in glutathione (GSH) biosynthesis, was increased 1.7-fold by β-NF. Changes in GCLC mRNA expression were paralleled by increases in intracellular GSH. In summary, largemouth bass hepatic CYP1A-dependent and GSH biosynthetic pathways, and to a lesser extent GST, are responsive to exposure to β-NF. 相似文献
8.
Mixed-function oxidase activity and conjugating enzymes in two species of antarctic fish 总被引：1，自引：0，他引：1
Silvano Focardi Cristina Fossi Claudio Leonzio Paolo Di Simplicio 《Marine environmental research》1989,28(1-4)
Enzyme activities of phase I and phase II of the hepatic detoxication system were investigated in two species of fish from the Ross Sea in Antarctica. The fish had extremely low phase I enzyme (BPMO and aldrin epoxidase). The reductase regenerating enzyme activities were comparable to those found in fish of temperate seas. Transferases of cytosol showed highest activity in the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and little or no conjugation of other substrates. The GSSG reductase and the glutathione transferase with peroxidase activity (CuOOH substrate) were low; the Sedependent glutathione peroxidase activity was not detectable. Interspecific differences were found for most of the enzymes tested. 相似文献
9.
Previous research had documented prevalent liver neoplasia within feral fish populations from half a dozen polluted waterways of North America.1–3 A brown bullhead, Ictalurus nebulosus, population in the Black River, Ohio, had a 33% prevalence of grossly observable liver tumors in fish older than 2 years compared to 0 % in 249 bullhead from a reference location (Buckeye Lake).4 To approximate the actual prevalence of gross plus microscopic lesions combined in the Black River population, we captured 125 bullhead at least 250 mm in length in 1982. Livers were preserved for histopathology and pectoral fin spines were removed for aging. On average, each liver was sectioned at five levels for diagnosis. Bullheads (whole fish) and Black River sediment were analyzed for polynuclear aromatic hydrocarbons (PAHs) by gas chromatography-mass spectrometry as previously reported.4,5 Bullheads were found to have a higher incidence of liver tumors than was grossly apparent and to contain elevated levels of polynuclear aromatic hydrocarbons (PAHs). 相似文献
10.
11.
Geracitano LA Bocchetti R Monserrat JM Regoli F Bianchini A 《Marine environmental research》2004,58(1):1-17
Acute (4 days) and chronic (14 days) effects of copper were evaluated on the antioxidant defenses of Laeonereis acuta (Polychaeta) collected in unpolluted (UP) and polluted (P) sites. In the acute assay (125 and 250 micro g Cu/l) superoxide dismutase (SOD) and glutathione S-transferase (GST) activities did not change, whereas catalase (CAT) increased in worms from both the sites. Lipid peroxidation was higher in copper exposed worms from the P site. In the chronic assay (62.5 micro g Cu/l) polychaetes from the P site showed enhanced activities of SOD, GST and CAT and higher contents of metallothionein-like proteins and sulfhydrils compared to worms from UP. Differences in responses between polychaetes from UP and P sites suggest that organisms from the polluted site, P, are more susceptible to oxidative stress conditions. 相似文献
12.
cDNA clones for glutathione S-transferases A (GST-A) and A1 (GST-A1) from plaice (Pleuronectes platessa) were expressed as N-terminally 6XHis tagged proteins in Escherichia coli and purified to homogeneity from Ni-NTA silica. GST-A was an efficient catalyst for conjugation of unsaturated alkenals derived from peroxidation of polyunsaturated fatty acids with the highest activity observed with trans-non-2-enal (8 micromol min(-1) mg(-1)). GST-A1 was a very efficient Se-independent glutathione peroxidase with an activity towards cumene hydroperoxide of 25 micromol min(-1) mg(-1). Although the enzymes exhibited moderately high activities towards the model substrate 1-chloro-2,4-dinitrobenzene (CDNB) they exhibited little or no activity towards other common prototypical xenobiotic substrates. Together with data for ontogeny, tissue distribution and inducibility of these enzymes, we contend that a primary function of these enzymes is protection from the harmful effects of lipid peroxidation products generated naturally or exacerbated by xenobiotic exposure. 相似文献
13.
Grant W. Tempero Brendan J. Hicks Nicholas Ling Dai Morgan Adam J. Daniel Deniz Özkundakci 《新西兰海洋与淡水研究杂志》2019,53(3):397-415
Introduced common carp (Cyprinus carpio) are a recognised threat to New Zealand’s freshwater ecosystems. In 2011, an invasive fish removal programme was undertaken in 16.8 ha Lake Ohinewai, a Waikato riverine lake. Active fish removal using netting and boat electrofishing was supported by installation of a one-way exclusion barrier on the outflow to Lake Ohinewai. The barrier was designed to allow fish passage downstream out of the lake, while preventing adult carp from moving upstream. The estimated carp population was reduced from 8,548 fish (5,863–12,937, 95% confidence limits) in 2011–454 fish (251–889, 95% CL) in 2014. However, by 2016 the carp population had recovered to 2,063 fish (1,070–4,328, 95% CL) with increased abundance in invasive brown bullhead catfish (Ameiurus nebulosus) also observed. This research demonstrates that carp populations can be reduced to low abundances in the short to medium term, but continued removal will be required for on-going control. 相似文献
14.
Seasonal variations in the antioxidant enzymes (catalase, superoxide dismutase [SOD], NADH-DT diaphorase), biotransformation enzyme, glutathione-S-transferase (GST) and microsomal lipid peroxidation in digestive tissue of barnacle, Balanus balanoides, from polluted and non-polluted populations have been evaluated. Relationships with accumulated polyaromatic hydrocarbon (PAH) concentration in barnacle tissues and environmental parameters (water temperature, salinity, dissolved oxygen concentration, water pH) were determined. As a general trend, maximum antioxidant enzyme and GST activities were detected in the pre-monsoon period or summer (March-June) followed by a gradual decrease during the monsoon (July October) with a minimum in the post-monsoon period or winter (November February). This pattern was similar to tissue concentrations of PAHs, resulting in a significant positive correlation with antioxidant enzymes, mainly catalase and SOD. Microsomal lipid peroxidation exhibited an almost reverse trend of seasonal variation to that of antioxidant enzyme activities indicating an enhanced susceptibility of barnacle tissues to oxidative stress. Among the environmental parameters, only water temperature seemed to have a significant effect on observed variations of antioxidant enzymes and GST activities. The barnacles from polluted and non-polluted populations exhibited seasonal differences in the activities of all the enzymes studied, particularly catalase, SOD and GST, suggesting the possibility of some biochemical adaptation in organisms from a chronically polluted environment. The results indicated that antioxidant defense components, catalase and SOD, are sensitive parameters that could be useful biomarkers for the evaluation of contaminated aquatic ecosystems. The results also suggested the potentiality of barnacle, B. balanoides, as a bioindicator organism against organic pollution. 相似文献
15.
GSTA is a major glutathione S-transferase gene responsible for 4-hydroxynonenal conjugation in largemouth bass liver 总被引：2，自引：0，他引：2
We have previously shown that largemouth bass (Micropterus salmoides) has a remarkable ability to conjugate 4-hydroxy-2-nonenal (4HNE), a mutagenic and cytotoxic alpha,beta-unsaturated aldehyde produced during the peroxidation of lipids. In addition, we have isolated a glutathione S-transferase cDNA (bass GSTA) that encodes a recombinant protein which is highly active in 4HNE conjugation and structurally similar to plaice (Pleuronectes platessa) GSTA. In the present study, HPLC-GST subunit analysis revealed the presence of at least two major GST isoforms in bass liver, with one peak constituting 80% of the total bass liver GST protein. Liquid chromatography mass spectrometry (LC-MS) and electrospray ionization analysis of the major bass GST subunit yielded a molecular weight of 26,396 kDa. Endo-proteinase Lys-C digestion and Edman degradation protein sequencing of this GST peak demonstrated that this protein was encoded by bass GSTA. Analysis of genomic DNA fragments isolated by nested PCR indicated the presence of a GST gene cluster in bass liver that contained GSTA, and was similar to a GST gene cluster characterized by Leaver et al., in plaice. Collectively, our data indicates the presence of a major GST in bass liver involved in the protection against oxidative stress. This GST is part of a gene cluster that may be conserved in certain freshwater and marine fish. 相似文献
16.
Richard T. Di Giulio Jeff V. Behar David B. Carlson Bruce M. Hasspieler David E. Watson 《Marine environmental research》1995,39(1-4)
Xenobiotic-mediated productions of reactive oxygen species, via enzymemediated redox cycling, have been implicated in a variety of toxicological phenomena including lipid peroxidation, enzyme inactivation and oxidative DNA damage leading to cancer. A comparison was undertaken of two benthic freshwater fish species that appear to differ markedly in their susceptibility to chemical carcinogenesis—the channel catfish (Ictalurus punctatus) and the more cancer-prone brown bullhead (Ameriurus nebulosus)-in terms of basic biochemical characteristics related to oxidative stress. This has included analysis of microsomal redox cycling of model xenobiotics (e.g. menadione) as well as antioxidant and other detoxifying enzymes in hepatic tissue of the two species. In addition, endpoints of oxidative stress, such as altered glutathione status and oxidative DNA damage, were examined. These studies have revealed numerous qualitative and quantitative differences between the two species both in terms of basal enzyme activities and in species response to model prooxidants. For example, bullhead appear to possess a greater capacity for microsomal redox cycling of xenobiotics, but have glutathione-dependent defense systems less able to withstand oxidative challenge. These and other interspecific differences have allowed for an improved understanding of the basic mechanisms which may underly species susceptibility to oxidative stress and critical manifestations such as cancer. 相似文献
17.
采用生物毒性测试与评价方法对常用消毒剂药物三氯异氰尿酸（Trichloroisocyanuric acid，TCCA）对剑尾鱼的急性毒性及其Ⅰ、Ⅱ相代谢酶活性影响进行了研究。结果表明，TCCA对剑尾鱼的96hLG0为2．15mg／L。TCCA对剑尾鱼肝脏还原型谷光苷肽（GSH）、谷胱苷肽硫转酶（GST）和7。乙氧基异吩恶唑酮-脱乙基酶（EROD）都存在诱导作用。雌雄个体在GST和EROD的诱导响应时间有显著差异性，其中雄性个体GST对TCCA暴露响应比雌性个体敏感。 相似文献
18.
Laeonereis acuta was seasonally collected in an industrially polluted site (P) and in an unpolluted site (UP) at the Patos Lagoon estuary (southern Brazil). Glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) activity (U/mg protein) was determined in five groups of worms from each site. Metallothionein (MT - mol GSH/g ww) and lipid peroxides content (LPO - nmoles of cumene hydroperoxide/g ww) were also measured. Annual mean values for CAT (UP=3.7+/-0.3; P=5.7+/-0.6), GST (UP=0.034+/-0.003; P=0.045+/-0.004) and MT (UP=0.15+/-0.02; P=0.23+/-0.03) were higher (p<0.05) in worms from the P site. In autumn, CAT activity was higher (p<0.05) in worms from the P site (7.6 +/- 1.3) than in those from the UP site (3.6 +/- 0.4). In summer, MT concentration was higher in worms from the P site (0.37 +/- 0.03) than in those from the UP site (0.19 +/- 0.01). No significant difference (p>0.05) in the LPO content was observed in worms from the different sites or collected in different seasons. These results indicate that worms from the polluted site showed higher antioxidant responses than those from the unpolluted site, sufficient to prevent oxidative damage in terms of LPO. 相似文献
19.
20.
谷胱甘肽硫转移酶是一类在细胞解毒和抗氧化过程中发挥重要作用的超家族酶系。采用cDNA末端快速扩增技术,从菲律宾蛤仔中克隆获得Rho型GST和微粒体型GST基因的全长序列(分别命名为VpGSTR和VpGSTMi)。序列分析显示,VpGSTR和VpGSTMi的cDNA全长分别为942bp和661bp,编码234和149个氨基酸。系统分析结果表明,菲律宾蛤仔GSTR与GSTMi在进化树上的位置与其分类所处的位置一致。荧光定量PCR结果发现,VpGSTR和VpGSTMi广泛分布于所检测组织中,且在肝胰腺组织中表达量最高。在鳗弧菌侵染后,VpGSTR和VpGSTMi基因表达量在24h内均表现出明显的上升趋势。上述结果表明,VpGSTR和VpGSTMi可能在菲律宾蛤仔抵御细菌侵染导致的氧化应激中发挥重要功能。 相似文献