Biochemical markers of oxidative stress in Perna viridis exposed to mercury and temperature

Oxidative damage and antioxidant properties have been studied in Perna viridis subjected to short-term exposure to Hg along with temperature (72h) and long-term temperature exposures (14 days) as pollution biomarkers. The elevated thiobarbituric acid reactive substances (TBA-RS) levels observed in gills and digestive gland under exposure to Hg, individually and combined with temperature, as also long-term temperature stress have been assigned to the oxidative damage resulting in lipid peroxidation (LPX). Increased activities of antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase (GST) both in gills and digestive glands under long-term exposures to temperatures are more prominent to heat rather than cold stress suggesting activation of physiological mechanism to scavenge the ROS produced during heat stress. Also decreased values of reduced glutathione (GSH) on long exposures to temperature stress indicate utilisation of this antioxidant, either to scavenge oxiradicals or act in combination with other enzymes, was more than its production capacity under heat stress. The results suggest that temperature variation does alter the active oxygen metabolism by modulating antioxidant enzyme activities, which can be used as biomarker to detect sublethal effects of pollution.     

光质对烟草叶片生长发育过程中抗氧化系统的影响

以云烟87植株为材料,通过覆盖白、红、黄、蓝、紫色滤膜获得不同光质,于大田条件下研究了光质对烟草叶片生长发育过程中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽过氧化物酶(GPX)、谷胱甘肽还原酶(GR)等抗氧化酶活性,抗氧化剂谷胱甘肽(GSH)和抗坏血酸(AsA)以及丙二醛(MDA)含量的影响.结果表明,在烟草植株第11片叶生长发育的7~70 d内,其抗氧化酶活性和抗氧化物质含量呈现先升高后下降的变化趋势.与白光(对照)相比,黄光诱导烟草叶片SOD、CAT、APX和GR活性升高,以及AsA和GSH含量增加;而红光诱导APX和GR活性上升,以及GSH和AsA含量升高;但紫光却使SOD、CAT、POD、GR和GPX活性下降,GSH和AsA含量降低,而蓝光则使所有抗氧化酶活性和抗氧化物质含量降低.紫光和蓝光处理的烟草叶片中MDA含量较高,而黄光和红光处理的则较低.总体而言,在大田条件下,相对红光和黄光而言,蓝光和紫光处理下的烟草叶片更容易发生光氧化胁迫.     

镉对长江华溪蟹肝胰腺抗氧化酶活力的影响

重金属对环境的污染已成为全球面临的首要问题之一,其中镉(Cd2 )是一种广泛存在的毒性污染物,能通过消化道和呼吸道进入生物体,对机体造成损伤(Zyadah and Abdel-Baky,2000)。研究表明,Cd2 可以通过Ca2 通道穿过细胞膜进入机体(Roesijadi and Robinson,1994),诱导产生大量自由基和活性氧(ROS),从而形成氧胁迫(Toppi andGabbrielli,1994;Hegedus et al.,2001)。ROS可以与体内脂质、蛋白质和核酸反应,导致脂质过氧化、细胞膜损伤并且影响多种酶的活力,对生物体造成威胁。由于在水生生态系统中生物富集污染物的作用明显,故相对于陆地生…     

Antioxidant responses of the Mediterranean mussel, Mytilus galloprovincialis, to environmental variability of dissolved oxygen

Physiological responses of Mytilus galloprovincialis against environmental dissolved oxygen partial pressure (pO(2)) variation were studied in terms of the modulated induction of the main antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) and selenium-dependent glutathione peroxidase (GPX). Field in vivo studies were performed at two sites of the Lagoon of Venice, characterized by different aquatic environmental conditions implying different pO(2). SOD and GPX are more active in gills, and their complementary role is discussed. CAT is more active in the digestive gland, where the enzyme dismutates H(2)O(2) derived from divalent reduction of O(2) performed by various oxidases in peroxisomes. Antioxidant enzyme activities are correlated with water dissolved oxygen (DO), especially in the gills. This tissue, because of its anatomical localization and its physiological role, responds to DO variations modulating the induction of the antioxidant enzymes as a protection mechanism against potential toxicity due to increases in ROS formation.     

镉对泥蚶抗氧化酶系统的影响

研究了不同浓度的重金属Cd²⁺对泥蚶抗氧化酶活性的影响。设置了四组镉的梯度(0、0.025、0.05和0.1 mg·L^-1,分别在第12、24、48、96和144 h对各组织取样,用泥蚶内脏团与鳃制成酶样,对非特异性免疫抗氧化酶SOD、CAT、GST、GPX进行指标测定。结果表明,在开始的12～24 h,中低浓度组SOD酶的活力与对照组相比有显著性地提高,高浓度组SOD酶的活力变化不显著,随着暴露时间的延长和Cd²⁺浓度的增加,随后至144 h,高中浓度组SOD酶的活力降至低于对照组水平(P<0.05);GPX和GST活力的变化与SOD变化相似,它们的活性在12～48 h的范围内达到顶峰,然后与对照组相比出现显著下降;中低浓度组的CAT酶活力在第144 h时与对照组相比无显著差异(P<0.05)。在一定程度的Cd²⁺胁迫会对泥蚶体内的抗氧化酶系统造成损伤。     

Cloning and mRNA expression of antioxidant enzymes in the Pacific oyster, Crassostrea gigas in response to cadmium exposure

Cadmium (Cd) is one of the most toxic heavy metal pollutants in the aquatic environment and can induce the formation of reactive oxygen species (ROS) that cause oxidative stress. In present study, we cloned catalase (CAT) and glutathione peroxidase (GPX) cDNA, and investigated its time- and dose-related effects of three Cd concentrations (0.01, 0.05 or 0.1 ppm) on mRNA levels of antioxidant enzymes (superoxide dismutase (SOD), CAT, GPX) in the gill and changes enzyme levels in the hemolymph of the Pacific oyster, Crassostrea gigas. The cDNA indentified encoded proteins of 516 and 244 amino acids corresponding to CAT and GPX, respectively. BLAST analysis from other species indicated that the residues essential to the enzymatic function of CAT and GPX proteins of C. gigas are highly conserved. Cd treatment significantly increased antioxidant enzyme mRNA expression in the gill in a time- and dose-dependent manner. The mRNA expression at 0.1 ppm Cd concentration increased up to 3 days (CAT, GPX) or 7 days (SOD) and then decreased by 7 days (CAT, GPX) or 11 days (SOD). Aspartate aminotransferase, alanine amintransferase and hydrogen peroxide (H(2)O(2)) concentrations levels increased significantly with exposure to 0.05 or 0.1 ppm Cd for 7 days. These results suggest that antioxidant enzymes play important roles in the physiological changes related to metabolism and cell protection that occur in Pacific oysters exposed to Cd.     

Superoxide dismutase, catalase and glutathione peroxidase activities in the brain of streptozotocin induced diabetic rats

Brain antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) levels were studied in the brains of early diabetic (72 hr) and long term diabetic (one month) rats. Diabetes was induced by injecting streptozotocin (50 mg/kg, i.p.) in citrate buffer. One group of diabetic rats was treated with insulin (1U/day/animal). The results indicate that early diabetic rats exhibit increased SOD and CAT activities with no alteration in the GPX activity. On the contrary, increased CAT decreased GPX activities with no alteration in the SOD activity, was noted in the long-term Diabetic rats. Insulin treatment reversed these alterations in both the groups. It can be concluded that, in diabetic condition antioxidant enzyme levels are elevated and insulin treatment attenuated these changes. Hence, diabetes mellitus, if left untreated, may initiate degenerative processes and other CNS complications due to accumulation of oxidative free radicals.     

硒对亚硝酸钠导致的草鱼肝细胞氧化损伤的保护作用

以草鱼(Ctenopharyngodon idella)肝细胞(L8824)为研究对象,设置对照组、亚硝酸钠暴露实验组、亚硒酸钠孵育实验组和亚硒酸钠孵育后亚硝酸钠暴露实验组,探讨亚硒酸钠对不同浓度亚硝酸钠诱导L8824细胞氧化损伤及凋亡的保护作用。结果显示,亚硝酸钠暴露能抑制L8824细胞贴壁,导致细胞凋亡率增加。亚硝酸钠暴露引致L8824细胞的谷胱甘肽过氧化物酶(GPX)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性降低;gpx、sod和cat基因表达下调(P<0.05), DNA损伤诱导转录物3(ddit3)和bcl-2相关X蛋白(bax)基因表达上调(P<0.05)。亚硒酸钠(10μmol/L)孵育L8824对细胞形态和凋亡率无显著影响,但GPX、SOD和CAT活性上升,gpx、sod、cat、核因子E2相关因子2(Nrf2)和Kelch样环氧氯丙烷相关蛋白-1(keap1)基因表达上调(P<0.05)。亚硒酸钠孵育后亚硝酸钠暴露实验组,细胞凋亡率、GPX、SOD和CAT活性较对照组无显著变化,但B淋巴细胞瘤-2(bcl-2)基因表达显著上调(P<0...     

Peroxisomal participation in the cellular response to the oxidative stress of endotoxin

Exposure to a sublethal dose of endotoxin offers protection against subsequent oxidative stresses. The cellular mechanisms involved in generating this effect are not well understood. We evaluated the effect of endotoxin on antioxidant enzymes in liver peroxisomes. Peroxisomes have recently been shown to contain superoxide dismutase (SOD) and glutathione peroxidase (GPX) in addition to catalase. Peroxisomes were isolated from liver homogenates by differential and density gradient centrifugations. Endotoxin treatment increased the specific activity of SOD and GPX in peroxisomes to 208% and 175% of control activity, respectively. These findings correlated with increases in peroxisomal SOD and GPX proteins observed by immunoblot. Although the quantity of catalase protein was increased when assessed by immunoblot analysis, the specific activity of catalase was decreased to 68% of control activity. Activation of catalase with ethanol only restored catalase activity to control levels suggesting that catalase had undergone irreversible inactivation. The observed increase in GPX activity may represent a compensatory mechanism triggered by accumulating H₂O₂. The data presented here suggest for the first time that mammalian peroxisomal antioxidant enzymes are altered during the oxidative injury of endotoxin treatment.     

Antioxidant defence enzyme activities in hepatopancreas, gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube

The aim of our study was to determine the activity of antioxidant defence (AD) enzymes: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and the phase II biotransformation enzyme glutathione-S-transferase (GST) in the hepatopancreas, the gills and muscle of Spiny cheek crayfish (Orconectes limosus) from the River Danube and to compare tissue specificities of investigated enzymes. Our results indicated that both specific and total SOD activities in the hepatopancreas were lower compared to the gills and muscle. Total SOD activity in the gills was lower with respect to that in muscle. CAT and GSH-Px (both specific and total) activities were higher in the hepatopancreas compared to those in the gills and muscle. In the gills the specific and total GR activities were higher than in the hepatopancreas and muscle. The specific and total GST activities were higher in the hepatopancreas compared with the gills and muscle. Our study represents the first comprehensive report of AD enzymes in tissues of O. limosus caught in the River Danube. The noted tissue distributions of the investigated AD enzyme activities most likely reflected different metabolic activities and different responses to environmental conditions in the examined tissues.     

苯并（a）芘对大弹涂鱼肝脏抗氧化酶活性影响的初步研究

在实验条件下,研究了不同浓度苯并(a)芘(BaP)暴露对大弹涂鱼肝脏内抗氧化酶-超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)和过氧化氢酶(CAT)活性的影响.结果表明,不同浓度的BaP暴露对抗氧化酶活性产生不同程度的影响.低浓度组(3μg     

植物谷胱甘肽过氧化物酶研究进展

氧化胁迫可诱导植物多种防御酶的产生, 其中包括超氧化物歧化酶(SOD, EC1.15.1.1)、抗坏血酸过氧化物酶(APX, EC1.11.1.11)、过氧化氢酶(CAT, E.C.1.11.1.6 )和谷胱甘肽过氧化物酶(GPXs,EC1.11.1.9)。它们在清除活性氧过程中起着不同的作用。GPXs是动物体内清除氧自由基的主要酶类,但它在植物中的功能报道甚少。最近几年研究表明, 植物体内也存在类似于哺乳动物的GPXs家族, 并对其功能研究已初见端倪。本文综述了有关GPXs的结构以及植物GPXs功能的研究进展。     

Antioxidant response of the bivalve Pinna nobilis colonised by invasive red macroalgae Lophocladia lallemandii

Invasive species represent a risk to natural ecosystems and a biodiversity hazard. The present work aims to determine the antioxidant enzyme response – superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), the phase II detoxifying enzyme – glutathione S-transferase (GST) – and markers of oxidative damage – thioredoxin reductase (TR) and malondialdehyde (MDA) – in gills and digestive gland of Pinna nobilis and to study the antioxidant response effects in the bivalve colonised by the invasive macroalgae Lophocladia lallemandii. Colonised specimens were collected in a control area without L. lallemandii and another area completely colonised by L. lallemandii. All enzyme activities were found to be present in gills and digestive gland, with some tissue differences. CAT and SOD activities were higher in gills than digestive gland, whereas GST activity and MDA levels were higher in digestive gland. The presence of L. lallemandii induced a significant increase in the activities of antioxidant enzymes in both gills and digestive gland, except for CAT activity in gills. GST and TR activities were also increased in both tissues, as well as the MDA concentration. We can conclude that the presence of L. lallemandii colonising P. nobilis induces a biological stress and oxidative damage to the fan mussel.     

抗氧化系统参与循环干旱锻炼提高烟草植株抗旱性的形成

对漂浮育苗的烟草幼苗进行控水-半萎焉-复水-恢复的循环干旱锻炼。结果表明,这种干旱锻炼能显著提高烟草叶片抗氧化剂谷胱甘肽（GSH）、抗坏血酸（AsA）的含量、还原型抗氧化剂在总抗氧化剂中的比例和抗氧化酶包括超氧化物歧化酶（SOD）、抗坏血酸过氧化物酶（APX）、过氧化氢酶（CAT）、愈创木酚过氧化物酶（GPX）、谷胱甘肽还原酶（GR）的活性,降低丙二醛（MDA）的含量。当烟草植株遭受后续的干旱胁迫时,与未锻炼的对照相比,干旱锻炼过的烟草植株能保持较高的GSH和AsA含量、还原型抗氧化剂在总抗氧化剂中的比例和抗氧化酶（SOD、APx、CAT、GPX和GR）活性,以及较低的MDA含量,表明这种循环干旱锻炼提高了细胞抗氧化能力,有助于缓解烟草植株由干旱引起的氧化胁迫及其所导致的伤害,最终提高其抗旱性。这些结果表明。抗氧化系统参与了循环干旱锻炼提高烟草植株抗旱性的形成过程。     

Effect of vitamin E and selenium on antioxidant enzymes in brain,kidney and liver of cigarette smoke-exposed mice

The present study was conducted to evaluate the protective effects of vitamin E and selenium (Se) application on alteration of antioxidant enzyme activities against cigarette smoking induced oxidative damage in brains, kidneys and liver of mice. Male mice (balb/c) were exposed to cigarette smoke and treated with Se and/or vitamin E. Glutathione transferase (GST), glutathione peroxidase (GPX), glutathione reductase (GRX), superoxide dismutase (SOD) and catalase (CAT) enzyme activities in mice brain, kidney and liver were measured spectrophotometrically. GST, GPX, GRX, SOD and CAT enzyme activities in the brains of smoke-exposed mice were found lower than the enzymes activities of control mice and Se-and vitamin E-treated mice at the end of the three and five months. Opposite to brain, enzyme activities in kidneys and livers of smoke-exposed mice were found higher than the enzymes activities of control mice and Se-and vitamin E-treated mice at the end of the three and five months. Activities of GST, GPX, GRX SOD and CAT in the livers, kidneys and brains of smoke-exposed mice were found statistically different (p < 0.01) compared to control mice and Se-and vitamin E-treated mice. Combined application of vitamin E and Se had an additive protective effect against changing enzymes activities in smoke-exposed mice livers, kidneys and brains at the end of the both application periods. These results suggest that cigarette smoke exposure enhances the oxidative stress, thereby disturbing the tissue antioxidant defense system and combined application of vitamin E and Se protects the brain, kidney and liver from oxidative damage through their antioxidant potential.     

Oxidative stress-inducible antioxidant adaptive response during prostaglandin F2alpha-induced luteal cell death in vivo

Oxidative stress-induced antioxidant adaptive response would be particularly important to cells in high reactive oxygen species (ROS) environments. We aimed to determine the dynamic adaptive response of antioxidant enzymatic systems in sheep corpus luteum (CL) during PGF2alpha-induced luteal cell death. Activities of superoxide dismutase (SOD1 and SOD2), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GSR), and in situ DNA fragmentation were determined in CL at day 10 of the estrous cycle (0 h) and at 12, 24 or 48 h after PGF2alpha injection. A decrease in plasma progesterone concentration was first observed at 6 h after treatment (P < 0.05). Apoptotic cells were rarely observed in the CL at 0 h (less than 0.7%), and their incidence increased (P < 0.01) by 12 h post-PGF2alpha (11.7%) and remained thereafter elevated through 48 h. Activities of SOD1, SOD2, GPX and GSR were not changed at any time points after PGF2alpha treatment. CAT activity increased at 12 h (P < 0.01) and at 24 h (P < 0.05) after PGF2alpha treatment as compared to that at 0 h. These findings demonstrate that PGF2alpha induce luteal cell death without depressing the activity of antioxidant enzymes. It is suggested that transient increase in CAT activity is an adaptive response of the CL to oxidative stress induced by PGF2alpha.     

镉对尖紫蛤抗氧化酶活性及脂质过氧化的影响

为阐明镉(Cd2+)对尖紫蛤消化盲囊和鳃抗氧化酶的毒性影响程度,研究了不同浓度的Cd2+(0.005、0.05、0.5 mg/L)在不同暴露时间(24h、72h、120h)对尖紫蛤鳃和消化盲囊中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)的活性以及丙二醛(MDA)含量的影响。结果表明,在Cd2+浓度为0.005 mg/L时,在整个实验期间Cd2+对消化盲囊和鳃内的SOD、CAT、GSH-PX活性并无显著影响。在Cd2+浓度为0.05 mg/L和0.5 mg/L时,SOD、CAT、GSH-PX在鳃和消化盲囊中的活性都呈现出明显的时间剂量依赖关系。在0.05 mg/L暴露时,鳃和消化盲囊中的SOD、CAT和GSH-PX的活性随时间逐渐增强,在72h时达到最大值,但在120h时略有降低。在0.5 mg/L暴露时,消化盲囊中SOD、CAT及鳃中CAT活性在24h时上升达到最大值,但鳃中SOD直到72h才达到最大值,并均在120h下降到最低,其中消化盲囊中SOD和CAT活性在120h低于对照组,这可能与消化盲囊对Cd2+的敏感性高于鳃有关。在0.5 mg/L暴露的鳃中,GSH-PX在24h、72h活性并不上升,在120h甚至低于正常水平,0.5 mg/L暴露的消化盲囊中,24h时迅速增高,然后逐渐下降到正常值。这可能与Cd2+结合了GSH-PX的活性中心,降低了GSH-PX的活性有关。与3种酶活性随着时间延长和剂量的增加,酶活性会降低的变化趋势不同,鳃和消化盲囊中的MDA的含量随时间延长和剂量的增加而增加,并不出现下降,这表明尖紫蛤鳃和消化盲囊中的MDA含量可以灵敏的反映机体内的氧化损伤程度,但不能敏感的反映水体中Cd2+的污染情况。     

Oxidative stress induced by intermittent exposure at a simulated altitude of 4000 m decreases mitochondrial superoxide dismutase content in soleus muscle of rats

The effects were examined of 6-month intermittent hypobaric (4000 m) exposure on the antioxidant enzyme systems in soleus and tibialis muscles of rats. At the end of the 6-month experimental exposure, the six rats in both the exposed group and the control group were sacrificed. Immunoreactive mitochondrial superoxide dismutase (Mn-SOD) contents were measured as well as the activities of antioxidant enzymes [Mn-SOD, cytosolic SOD (Cu,Zn-SOD), catalase (CAT), and glutathione peroxidase (GPX)]. Thiobarbituric acid-reactive substances (TBARS) were also determined as an indicator of lipid peroxidation. The high altitude exposure resulted in a marked increase in TBARS content in soleus muscle, suggesting increased levels of oxygen free radicals. Conversely, significant decreases in both Mn-SOD content and activity in solens muscle were oted affer exposure. Such trends were not noticed in tibialis muscle. On the other hand, no significant changes in Cu,Zn-SOD, CAT, or GPX were observed in either muscle. These results suggested that the increases in lipid peroxidation were most probably a result of decreased Mn-SOD function which was more depressed in oxidative than in glycolytic muscle.     

Pro-oxidant and antioxidant processes in gas gland and other tissues of cod (Gadus morhua)

Partial reduction of molecular oxygen produces reactive oxyradicals, including the superoxide anion radical (O _- ² ) and hydroxyl radical (·OH). The gas gland functions under hyperoxic and acidic conditions and therefore is likely to be subjected to enhanced oxidative stress. Aspects of pro- and antioxidant processes in gas gland were compared with other tissues likely to be subject to differing degrees of oxyradical production, viz. liver (site of chemically-mediated oxyradical production), gills and skeletal muscle. Antioxidant enzyme activities (superoxide dismutase, catalase, selenium-dependent and total glutathione peroxidase) per g wet weight were highest in liver and lowest in muscle. Catalase and glutathione peroxidase activies per g wet weight were higher in gills than in gas gland, whereas the reverse was seen for superoxide dismutase. Cytosolic superoxide dismutase activities per mg protein were two- and nine-fold higher in gas gland than in liver and gills. The pH characteristics of the antioxidant enzymes were generally similar in all the tissues. Glutathione, vitamin E and unsaturated (peroxidizable) lipid levels were generally highest in liver followed by gas gland. Lipid peroxidation (malonaldehyde equivalents) was evident in all tissues except gas gland. Hydrogen peroxide and O _- ² were involved in the NAD(P)H-dependent ferric/EDTA-mediated formation of ·OH (as measured by 2-keto-4-methiolbutyrate oxidation) by mitochondrial and postmitochondrial fractions of gas gland. Tissue maximal potentials for ·OH production paralled superoxide dismutase but not catalase or glutathione peroxidase activities. Overall, the results confirm the presence of effective antioxidant defences in gas gland and support previous workers' contentions of a central role for superoxide dismutase in this process.Abbreviations EDTA di-sodium ethylenediaminetetra-acetic acid - G-6-P glucose-6-phosphate - GPX total glutathione peroxidase - GSH reduced glutathione - GSSG oxidised glutathione - GST glutathion-S-transferase - HPLC high performance liquid chromatography - KMBA 2-keto-4-methiolbutyric acid - MOPS 3-[N-morpholino] propane-sulphonic acid - PMS postmitochondrial supernatant - Se-GPX selenium-dependent glutathion peroxidase - SOD superoxide dismutase - TCA trichloroacetic acid