Antioxidant enzymes in the digestive gland of the common musselMytilus edulis

Antioxidant enzymes function to remove deleterious reactive oxygen species, including the superoxide anion radical and H₂O₂. Subcellular distributions and optimal and other properties of catalase (EC. 1.11.1.6), superoxide dismutase (SOD; EC. 1.15.1.1), selenium-dependent glutathione peroxidase (Se-GPX; EC. 1.11.1.9) and total glutathione peroxidase (GPX) activities were determined in the digestive gland of the common musselMytilus edulis L. by spectrophotometric and cytochemical/electron microscopic (catalase) techniques. Assay conditions for Se-GPX and total GPX activities were determined which optimized the difference between the non-enzymic and enzymic rates of reaction. General peroxidase activity (guaiacol as substrate) (EC. 1.11.1.7) was not detectable in any subcellular fraction. Catalase was largely, if not totally, peroxisomal, whereas SOD and GPX activities were mainly cytosolic. Distinct mitochondrial (Mn-SOD) and cytosolic (CuZn-SOD) SOD forms were indicated. Catalase properties were consistent with a catalase, rather than a catalase-peroxidase. The pH-dependence and temperature-dependence of GPX activity were different with H₂O₂ or CHP as substrate, and these and other observations indicate the existence of a distinct Se-GPX. Under saturating or optimal (GPX) assay conditions, the apparent Michaelis constantsK _m (mM) were: catalase, 48 to 68 (substrate, H₂O₂); Se-GPX, 0.11 (H₂O₂) and 2.0 (glutathione); and total GPX, 2.2 (eumene hydroperoxide) and 1.2 (glutathione). Calculated catalase activity was 2 to 4 orders of magnitude greater than Se-GPX activity over an [H₂O₂] of 1 to 1000 M. The results are discussed in relation to theoretical calculations of in vivo oxyradical production and phylogenetic differences in antioxidant enzyme activities.     

Zn2+胁迫及恢复下番茄幼苗根系中抗氧化系统的动态变化

通过温室水培试验,研究1.0×10-4mol/LZn2 胁迫及解除胁迫对番茄(Lycopersicom esculentum Mill.)幼苗根系中抗氧化系统的动态变化.结果表明,胁迫条件下,番茄根系丙二醛(MDA)和蛋白质含量增加;超氧化物歧化酶(SOD,EC1.15.1.1),过氧化氢酶(CAT,EC1.11.1.6)和谷胱甘肽过氧化物酶(GPX,EC1.11.1.9)活性上升,而过氧化物酶(POD,EC1.11.1.7)活性下降;AsA-GSH循环代谢受影响;抗坏血酸过氧化酶(APX,EC1.11.1.11)活性在胁迫d1、d3上升,随后下降,而谷胱甘肽还原酶(GR,EC1.6.4.2)活性在胁迫1~5d时下降,随后上升;抗坏血酸(AsA)和谷胱甘肽(GSH)含量上升.解除胁迫后,根系中MDA和蛋白质含量随时间延长逐渐减少,接近对照;POD和APX活性增加,而SOD、CAT和GR活性及GSH含量先升高,随后减少;GPX活性和AsA含量却降低;恢复后根系中抗氧化物水平仍然高于对照.依据实验结果,文中讨论了胁迫及恢复过程中植物AsA-GSH循环代谢的作用.图3参16     

Effect of ambient oxygen concentration on activities of enzymatic antioxidant defences and aerobic metabolism in the hydrothermal vent worm, Paralvinella grasslei

The alvinellid Paralvinella grasslei is a common endemic polychaete from the deep-sea hydrothermal vent communities located on the East Pacific Rise (EPR). These organisms colonise a large range of microhabitats around active sites where physico-chemical conditions are thought to generate reactive oxygen species (ROS). Furthermore, in this aerobic organism, ROS could also be generated by the activity of the mitochondrial respiratory chain. In this paper, we investigated the effect of ambient oxygen concentration on the activities of three essential antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX) and their relationships with the activity of enzymes involved in aerobic metabolism (cytochrome c oxidase, COX; citrate synthase, CS). Results of incubation of P. grasslei in a high-pressure vessel with circulating seawater at different oxygen partial pressures indicate that this worm regulates COX and CS activities differently in gills and body wall. CAT and GPX activities increase in these tissues when animals are maintained in filtered surface seawater. Moreover, levels of malondialdehyde increase in gills, testifying that oxidative damage occurs under these conditions. CAT and GPX activities are positively related to COX and CS activities, but no correlation was detected between SOD and the metabolic enzyme activities. In comparison with littoral annelids, SOD activities are very high whereas CAT activities are very low or absent in P. grasslei. The possible reasons for the occurrence of such differences are discussed.     

Identification of haemocytes and their role in clotting in the blue crab,Callinectes sapidus

Three types of haemocyte — hyaline cells, small granule and large granule haemocytes — have been identified in the circulation of blue crabs, Callinectes sapidus Rathbun, that were collected and processed in North Carolina, USA in February 1991. Differentiation of haemocytes begins in the haematopoietic tissue and continues in the circulation. Hyaline cells are distinguished morphologically by their relatively small size, high nucleocytoplasmic ratio and sparse cytoplasmic granule content. Small and large granule haemocytes are distinguished, respectively, by a preponderance of small and large electron dense granules in their cytoplasm. Furthermore, a light and electron microscopic examination of the clotting process performed in July 1991 revealed that hyaline cells initiated coagulation when exposed to seawater, whereas granular haemocytes did not undergo marked changes when so treated. These distinct functional and morphologic features suggest that the hyaline cell and granular haemocytes represent distinct cell lines. It remains unclear whether small granule haemocytes differentiate into large granule haemocytes.     

First evidence of gender-related differences in immune parameters of the clam Ruditapes philippinarum (Mollusca, Bivalvia)

For the first time, gender-related differences in some important functional parameters of haemocytes from the clam Ruditapes philippinarum have been studied. Clams were collected during the pre-spawning phase, when sex can be distinguished by microscopic observation of gonadal smears, in order to ascertain whether the two sexes reach the stressful spawning period with different degrees of immunosurveillance. Total haemocyte count (THC), haemocyte size frequency distribution, capability of haemocytes to assume the vital dye neutral red (NR, indicative of endocytotic activity), lysozyme-like and acid phosphatase activities in both haemocyte lysate (HL) and cell-free haemolymph (CFH), and superoxide dismutase (SOD) and catalase (CAT) activities in HL were all evaluated. No differences in THC values were found between the two sexes, but differing haemocyte size frequency distributions were observed: the fraction of larger haemocytes (7–9 μm diameter, 200 fl volume) was markedly increased in females and that of smaller haemocytes (<5 μm diameter, <200 fl volume) markedly increased in males. Interestingly, when haemocytes were observed under the light microscope, a significantly higher percentage of granulocytes was found in haemolymph from females, compared with that of males. Significantly increased NR uptake was recorded in female haemocytes. No significant variation in lysozyme-like activity was observed in HL, whereas in CFH, enzyme activity was significantly higher in females. HL acid phosphatase activity was significantly higher in males with respect to females; no significant variations in enzyme activity were observed in CFH. HL SOD and CAT activities were significantly higher in females than in males. Overall, on the basis of the cell parameters studied here, gender-related differences in immune parameters were found in R. philippinarum, indicating that females have more active haemocytes than males during the pre-spawning period.     

Screening of endogenous antioxidants in some medicinal plants

The evaluation of antioxidant enzymes and antioxidant metabolites was done in leaf tissues of Azadirachta indica, Butea monosperma, Cassia fistula, Mangifera indica, and Syzygium cumini growing in the Thar Desert, Rajasthan, India. The plants are naturally exposed to drought stress and high temperatures during summer. Enzymatic reactive oxygen species (ROS) scavenging mechanisms in plants include superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase, catalase, and guaiacol peroxidase, and non-enzymatic antioxidants including the carotenoids, proline, and vitamin C were studied. The strategies to cope up with ROS under these extreme conditions are plant-specific. The highest activity of APX was found in M. indica (13.6?±?2.4?units?g^?1 fresh wt.). A. indica exhibited maximum guaiacol peroxidase activity (0.024?±?0.006?units?min?g^?1 fresh wt.), while S. cumini showed maximum SOD (12.5?±?2.3?units?g^?1 fresh wt.) and catalase activities (6.9?±?2.2?units?g^?1 fresh wt.). M. indica and S. cumini have been found to be more potent antioxidant systems among the studied plants.     

Antioxidative response of Lemna polyrrhiza L. to cadmium stress

Growth, lipid peroxidation, different antioxidative enzymes and metal accumulation were studied in Lemna polyrrhiza treated with different concentrations (1-40 ppm) of CdSO4. The growth of the plant was slightly enhanced with 1 ppm, while higher concentrations retarted growth and multiplication of fronds, the effect being concentration and dose dependant. Increase in malondialdehyde content was insignificant after the first week but a prolonged exposure led to significant (p < 0.05) increase of about 38% and 45% over the control in 20 and 30 ppm, respectively after four weeks. Catalase (EC 1.11.1.6; CAT) activity increased at low concentration, but it declined to 42% and 54% at 40 ppm after 6 and 30 days, respectively Superoxide dismutase (EC 1.15.1.1; SOD), ascorbate peroxidase (EC 1.11.1.11;APx) and glutathione reductase (EC 1.6.4.2) increased at both low as well at high concentrations, but a prolonged exposure to high concentration of Cd (40 ppm) led to significant (p < 0.05) decline in the mean activities of these antioxidant enzymes. Accumulation of Cd in biomass was concentration and time dependant However at high concentration of 40 ppm, Cd accumulation did not increase significantly (p < 0.05) with time. Increased activities of antioxidant enzymes in Cd treated plants suggest that metal tolerance in L. polyrrhiza might be associated to the changes of antioxidant enzymatic activities.     

Impact of environmental concentrations of beta-cypermethrin on the antioxidant system in the brain and liver of zebrafish (Danio rerio)

Beta-cypermethrin (beta-CYP) is a widely used pyrethroid pesticide, the extensive application of which may potentially cause damage to non-target organisms. To investigate the effect of beta-CYP on the antioxidant system of aquatic animals, adult zebrafish were exposed to environmentally relevant dosages (0.01, 0.1 and 1.0 μg/L) of beta-CYP. The activities of four antioxidant enzymes in zebrafish liver and brain tissue were tested after 7, 15 and 30 days of exposure. Our results showed that exposure of beta-CYP could induce different levels of increase in hepatic superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx) activities at 7 and 15 days post exposure (dpe), but caused apparent inhibition of hepatic SOD, GR and GPx activities at 30 dpe. Unlike in liver tissue, SOD and CAT activities in zebrafish brain did not show any apparent response to beta-CYP during the whole treatment period. In addition, increased brain GPx activities were observed at 7 and 30 dpe.     

Protective role of L-ascorbic acid against cypermethrin-induced oxidative stress and lipid peroxidation in Wistar rats

Cypermethrin is a synthetic pyrethroid insecticide used for pest control in agriculture and as an acaricide in man and animals. This study was undertaken with the objective to investigate the propensity of cypermethrin to induce oxidative stress in rats following repetitive dermal exposure and its possible attenuation by L-ascorbic acid. Results obtained showed that cypermethrin significantly (p < 0.05) increased malonaldehyde levels, activity of catalase in rat erythrocytes and plasma protein levels. Whereas, activities of glutathione (GSH), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were significantly (p < 0.05) reduced in the cypermethrin exposed rats as compared to the control. Supplementation of L-ascorbic acid in cypermethrin-exposed rats decreased lipid peroxidation of erythrocytes, total plasma protein and catalase activity significantly (p < 0.05) compared to non-cypermethrin-exposed group. However, L-ascorbic acid did not alleviate the negative effects of cypermethrin on the activities of SOD and GSH. This study revealed that the presence of L-ascorbic acid diminishes the adverse effects of cypermethrin on some oxidative stress parameters.     

芘暴露对马氏珠母贝鳃和肝胰腺抗氧化酶活性的影响

在实验生态条件下,采用生态毒理学方法,选用海洋环境中常见的有机污染物多环芳烃芘（Pyrene）,以马氏珠母贝（Pinctada martensi）为实验材料进行毒理实验。研究了芘不同质量浓度（8、16、32和64μg·L^-1、不同时间（7,10 d）的胁迫对马氏珠母贝鳃和肝胰腺组织中超氧化物歧化酶（SOD）、过氧化物酶（POD）和谷胱甘肽过氧化物酶（GPx）3种抗氧化酶活性的影响。结果表明：3种酶在不同组织和不同胁迫时间的敏感性有所差异,在芘胁迫马氏珠母贝第7天,鳃组织中 SOD、POD、GPx 3种酶活性主要表现为诱导现象,高质量浓度组对3种酶活性均具有极显著诱导作用（p〈0.01）,而芘胁迫对肝胰腺3种酶活性的影响主要表现为抑制作用,且POD酶活性表现最敏感,在较低浓度组就被显著抑制（p〈0.01）;到染毒第10天,鳃组织中SOD、GPx两种酶诱导作用明显,POD诱导现象不显著（p〉0.05）,肝胰腺中3种酶活性与对照组相比诱导明显,并表现出一定的剂效关系。两种组织中SOD和GPx活性的变化具有一定的同步性。通过比较最低可观察效应质量浓度（LOEC）,发现在第7天,鳃组织中敏感性强弱表现为SOD=POD〉GPx,肝胰腺中表现为POD〉GPx〉SOD,而到第10d,两组织中敏感性又分别表现为GPx〉SOD〉POD,GPx=SOD〉POD。因此,在较短的暴露时间,POD酶活性可作为芘胁迫对马氏珠母贝的生物标志物,而对于较长时间的染毒实验,可选用 GPx 酶作为芘胁迫对马氏珠母贝的生物标志物以监测海洋环境芘污染。     

硫酸锰废渣浸出液对斑马鱼的毒性及抗氧化酶活性的影响

已有研究表明,堆积在湘西花垣河河岸的硫酸锰废渣含有多种重金属,可能是导致花垣河河水和沉积物重金属污染的主要原因.为了评价硫酸锰废渣的潜在生态风险和对水生动物的毒性,本研究分析了硫酸锰废渣浸出液中的主要重金属成分,用漫出液对斑马鱼进行了急性毒性实验,观察在急性胁迫下斑马鱼的体征变化,测定了斑马鱼在亚致死浓度下的生化指标的...     

Study of the effect of refinery waste water exposure on Allium cepa's enzymatic antioxidant defense: probing potential biomarkers

The effects of heavy metals, hydrocarbons, and various toxicants present in Mathura Refinery Waste Water (MRWW) on Allium cepa were examined as a model plant system. The study was based on exposure of A. cepa to different concentrations of MRWW and compared to untreated control (exposure of onion bulbs with aquaguard purified water) to determine the total protein content and activities of certain antioxidant enzymes. These enzymes were evaluated for their efficacy to serve as biomarkers of refinery waste water pollution. DNA damaging potential of MRWW was also investigated. Data demonstrated maximal enhancement in ascorbate peroxidase activity subsequent to MRWW insult, although a significant increase in activities was also noted for other enzymes in the following order: superoxide dismutase?>?glutathione-S-transferase?> catalase. This is suggestive of their potency as a biomarker of MRWW toxicity. Increase in activities of monodehydroascorbate reductase (MDHAR; 152%) and deoxyribonuclease (345%) were also found in the A. cepa system as a result of MRWW exposure. In conclusion, A. cepa system might thus serve as an appropriate tool for monitoring water pollution, especially produced by petroleum waste and heavy metals in term of induction of glutathione peroxidase, MDHAR, and deoxyribonuclease activity.     

Studies on the role of six enzymes in the metabolism of kinetin in mustard aphid, Lipaphis erysimi (Kalt.)

The activity of catalase, glutathione peroxidase, superoxide dismutase, O-demethylase, ATPase and succinate dehydrogenase, belonging to two main classes of detoxification enzymes (i.e. hydrolases and oxido-reductases), mostly involved in metabolism and degradation of xenobiotics in insects, were assessed under the influence of kinetin, a plant growth regulator (PGR). The nymphs (48-52 hr old) of Lipaphis erysimi (Kalt.) were permitted to feed on radish plant, Raphanus sativus L. treated with kinetin (400 ppm) for 13, 25 and 37 hr. It was found that the activity of catalase, glutathione peroxidase and superoxide dismutase increased significantly when compared with the control of the same age group, which indicated that these enzymes might be playing a significant role in the metabolism of kinetin in this insect. The activity of O-demethylase showed an increase up to 25 hr of the treatment but it decreased under prolonged treatment whereas the activity of succinate dehydrogenase fluctuated insignificantly. ATPase showed a decrease in the activity with the treatment suggesting kinetin's interference in synthesis of ATPase.     

Responses of the earthworm Eisenia andrei exposed to sublethal aluminium levels in an artificial soil substrate

Industrial discharges of Al are increasingly common. In this study, the activities of three antioxidases, malondialdehyde (MDA) content and changes in coelomocytes were investigated in earthworms (Eisenia andrei) exposed to different concentrations of aluminium chloride (range 0–200 mg Al kg^?1 dry soil) in artificial soils for 32 days. Within the first 16 days of Al exposure, catalase activity was elevated in most treatments, while the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were inhibited in all treatments. After 16 days exposure, SOD and GSH-Px activities returned to normal at lower Al concentrations, but were still inhibited at higher concentrations, hence the inhibition of SOD and GSH-Px showed some exposure-level dependence during the late exposure phase. MDA content was significantly elevated from 16 days after initial exposure. Neutral Red retention time of coelomocytes decreased at the highest exposure concentrations after 32 days, and an increased proportion of cells showing membrane damage was also observed at this time using flow cytometry. We conclude that excessive Al in soils can cause oxidative stress and cell damage in earthworms. Hence, Al accumulation in the environment may present an ecological hazard through suppression of the important functions of earthworms in soil ecosystems.     

Antioxidant enzymatic activities and lipid peroxidation in liver and ovary of zebrafish (Danio rerio) exposed to deltamethrin

Deltamethrin (DM) is being used as a substitute for organochlorines and organophosphates in pest control because of its low environmental persistence and toxicity. But it has become an environmental contaminant as it has been used widely. In this study, we investigated the effect of DM (technical grade) on the antioxidant system of adult zebrafish. For this, six-month-old fish were exposed to 2, 4 and 6?μg/L of DM for 96?h. The tissues selected were liver and ovary. Our data showed that exposure to DM increases CAT (catalase), SOD (superoxide dismutase), GPx (glutathione peroxidase, antioxidant enzymes), LPO (lipid peroxidation, non-enzymatic antioxidant) and GST (glutathione S-transferase, detoxifying enzyme) in liver and ovary. Increased GST could detoxify the toxicant; still there could be enough DM to cause oxidative stress. It appears from our study that zebrafish used compensatory mechanisms in eliminating reactive oxygen species. These data will be useful as oxidative stress is being used as a biomarker for aquatic pollution.     

乙酸铜对银鲳幼鱼急性毒性及抗氧化酶活性的影响

为评价乙酸铜对银鲳的安全性及毒性效应,采用静水急性暴露实验,研究了8个乙酸铜浓度梯度(0、0.150、0.206、0.282、0.387、0.531、0.729、1.00 mg·L~(-1))对银鲳幼鱼的急性毒性,以肝脏和鳃组织的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化酶(GSH—PX)以及谷胱甘肽巯基转移酶(GST)活性为指标研究乙酸铜对银鲳幼鱼的毒理作用,并进行安全评价。结果表明,高浓度乙酸铜会对银鲳幼鱼产生了较大的毒性,48 h和96 h-LC_(50)分别为0.898 mg·L~(-1)、0.264 mg·L~(-1),安全质量浓度为0.026 mg·L~(-1);不同浓度乙酸铜胁迫下银鲳幼鱼组织中的SOD、CAT、GSH-PX和GST活性均表现为低浓度被诱导而高浓度受抑制的规律,与乙酸铜浓度呈抛物线型剂量效应关系,此外,肝脏中4种抗氧化酶活性普遍高于鳃组织。研究发现,乙酸铜胁迫对抗氧化酶的影响,可以反映银鲳幼鱼机体的受损状况,其中SOD可较灵敏地指示早期低浓度的铜污染。     

Effects of copper on lipid peroxidation,glutathione, metallothionein,and antioxidative enzymes in the freshwater mussel Anodonta anatina

Copper is an essential element to all animals. At elevated concentrations, it is toxic and can participate in the formation of reactive oxygen species, leading to cellular damage. In this study, the ecotoxicological relevance of copper was investigated with freshwater mussels, Anodonta anatina. When the mussels were exposed to copper at environmentally realistic concentrations, either via the water (0.3?µmol?L^?1 Cu) or fed with Cu-loaded algae (equivalent to 0.06?µmol?L^?1 Cu), the level of thiobarbituric acid-reactive substances rose and glutathione decreased. This was associated with the induction of metallothionein and, relative to total protein, of glutathione reductase and the antioxidative enzymes superoxide dismutase, catalase, and glutathione peroxidase. But, since the overall protein-synthetic capacity was hampered by the copper insult, the activities of the enzymes relative to tissue weight and copper concentrations were depressed. During depuration, most parameters started to normalize although not returning to control values within 12 days.     

Mercuric chloride induced proteotoxicity and structural destabilization in marine fish (Therapon jarbua)

The reaction byproducts derived from lipid peroxidation (LPO), as well as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) enzyme activities were measured in tissues of marine fish (Therapon jarbua) exposed to mercuric chloride (HgCl) in water dispersions of 0.125 or 0.25 ppm. LPO was significantly increased in various tissues relative to control values after 96-h exposure. SOD and GPx activities significantly decreased after exposure to first two doses but significantly elevated CAT in Dose II kidney and liver tissues. The elevated levels of LPO, decreased activities of SOD and GPx, and increased CAT activities in all tissues examined in T. jarbua are an index of oxidative stress in fish. Structural analysis by scanning electron microscopy studies revealed the structural deformation in HgCl₂-exposed animals. These observations suggest that HgCl acts as a mediator in free radical generation. The increase in CAT and decrease in SOD and GPx activities in these tissues may be an adaptive response.     

Effects of mercury graded doses on redox status,metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali

Mercury (Hg) is a ubiquitous and a major environmental metal pollutant in the aquatic ecosystem. The present study was performed to evaluate the effect of mercury graded doses exposure on oxidative stress, redox status, metallothionein levels and genotoxicity in the intestine of sea cucumber Holothuria forskali. Specimens were exposed for 96?h to three concentrations of Hg (40, 80 and 160?µg/L). Exposure of H. forskali to Hg promoted oxidative stress with an increase in malondialdehyde (MDA), protein carbonyl (PCO) and advanced oxidation protein products (AOPP) levels. An increase of glutathione (GSH), vitamin C (ViteC) and non-protein thiols (NPSH) contents was also observed. Additionally, antioxidant activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) increased especially with the highest doses of Hg indicating a great defense of the antioxidant system. Our investigation revealed an increase in total Metallothionein (MT) content which was more pronounced at the sharpest concentrations of Hg. A significant decline of AChE activity was also observed. In the intestine of Hg-treated H. forskali genotoxicity was confirmed by DNA degradation.