昆虫体内抗氧化系统研究进展

昆虫为了减轻和防止活性氧损伤,已形成了复杂的氧化应激机制。可通过酶促如超氧化物歧化酶、过氧化物酶、过氧化氢酶等和非酶促谷胱甘肽、抗坏血酸和胡萝卜素等清除活性氧的系统以清除过量的活性氧。本文论述了昆虫在氧化胁迫下所具有的一套抗氧化系统。对其系统组成抗氧化酶和抗氧化剂的主要成分的抗氧化活性进行了综述。     

水分胁迫下植物体内OH的产生与细胞的氧化损伤

干旱是植物组织的一种重要的胁迫因子。它能干扰植物细胞中活性氧产生与清除的平衡,导致植物细胞遭受氧化胁迫。过去十多年来,人们对水分胁迫下植物体内活性氧的产生、活性氧对植物的伤害及植物保护系统的作用进行了大量的研究,取得了明显的进展［１～４］。然而,水分...     

植物谷胱甘肽过氧化物酶(GPX)研究进展

逆境胁迫会诱导植物产生过多的活性氧(ROS),引起氧化胁迫,严重影响其生长发育。相应地,植物为适应诸多不良环境会产生多种抗氧化剂、抗氧化酶等协调氧化还原平衡,其中谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)是植物体内重要的抗氧化酶之一。对近年来植物中GPX的结构、亚细胞定位、酶催化底物特点及作用研究进展进行综述,并对未来可能的研究方向进行展望。     

活性氧清除系统对干旱胁迫的响应机制

干旱胁迫是影响植物生长发育的主要因子,干旱引起活性氧自由基增加,使植物细胞遭受氧化胁迫．植物体通过酶促和非酶促两大保护系统清除活性氧,活性氧自由基的变化也会引起抗氧化防御系统的不同变化．同时干旱胁迫下活性氧的产生也与ABA的积累、脯氨酸的积累以及叶绿素荧光猝灭密切相关,因此了解活性氧清除系统对干旱胁迫的响应机制以及活性氧在植物生理生化过程中的作用是非常必要的。     

植物中的活性氧研究概述

活性氧作为有氧代谢的副产物不断在植物体内产生。在正常的生长环境条件下,植物将产生活性氧(reaction oxygen species, ROS)作为信号代谢分子以调控不同的代谢反应,例如病毒防御、细胞程序性死亡和气孔开闭等;当氧化胁迫发生时,胞内活性氧稳态受到严重破坏,影响作物的生长发育,从而降低作物产量及品质。为了降低因过量活性氧对植物体所造成的伤害,植物体内进化出了两种活性氧清除系统:酶清除系统和非酶清除系统。本文就此对植物在生长发育过程中ROS的产生、利弊、清除机制以及在作物改良上应用的可能性进行了系统的讨论。     

转铜/锌超氧化物歧化酶和抗坏血酸过氧化物酶基因马铃薯的耐氧化和耐盐性研究

高盐等逆境可以加剧植物体内活性氧的产生,进而引起植物细胞死亡。为开发抗逆境作物,以置于氧化诱导型启动子下定位于叶绿体的转铜/锌超氧化物歧化酶（Cu/ZnSOD）和抗坏血酸过氧化物酶基因(APX)马铃薯为材料,研究了其对MV和 NaCl所引起的氧化胁迫的耐受性。结果表明, MV胁迫下,转基因马铃薯叶片膜的相对电导率明显低于对照; NaCl胁迫下,其叶绿素含量高于对照。 在含NaCl 的培养基上,转基因幼苗生根率明显大于对照。另外,NaCl胁迫下转基因马铃薯叶片的SOD和APX酶活性显著高于对照,与其耐盐性的提高相一致。这些研究表明,转入Cu/ZnSOD和APX基因的马铃薯清除活性氧的能力增强,抗逆性得到提高。本实验采用氧化诱导型启动子调控下的SOD和APX两个基因协同作用,使外源基因只有在逆境胁迫时才特异性表达,增强转基因植株的抗逆效果,为培育抗逆经济作物开阔了思路。     

钙对根际低氧胁迫下黄瓜幼苗活性氧代谢的影响

采用营养液栽培系统,以黄瓜品种中农8号为材料,研究了Ca2 对根际低氧胁迫下黄瓜幼苗体内超氧阴离子(O2?-)、过氧化氢(H2O2)、丙二醛(MDA)含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱苷肽还原酶(GR)活性的影响.结果表明:低氧胁迫下黄瓜体内活性氧含量和保护酶活性均高于对照;低氧缺钙处理的活性氧含量最高,而保护酶活性却较低;营养液Ca2 浓度提高到8mmol/L后,显著降低了低氧胁迫下黄瓜幼苗体内MDA、H2O2含量和O?2-产生速率,提高了SOD、POD、CAT、APX、GR活性,说明Ca2 可减少低氧胁迫下黄瓜幼苗体内活性氧的产生,提高抗氧化酶的活性、降低膜脂过氧化水平,减缓低氧胁迫对植株的伤害,增强黄瓜幼苗对低氧逆境的适应性.     

不同活性氧胁迫下Bacillus sp. F26以抗氧化物酶合成为特征的应激响应

采用不同的活性氧发生源, 研究了· 、H2O2和OH·胁迫下Bacillus sp. F26以抗氧化物酶合成为特征的应激响应。结果表明, 细胞对氧胁迫的应激响应程度取决于活性氧种类、胁迫程度和形式(瞬时和持续)。Bacillus sp. F26对H2O2胁迫的响应程度最高, 过氧化氢酶的快速合成对细胞抵抗H2O2胁迫至关重要, 当细胞及时分解进入胞内的H2O2, 胁迫对细胞的氧化损伤程度并不高, 相反会刺激细胞的生长和底物消耗, 当胁迫超过过氧化氢酶的分解能力时, H2O2会迅速抑制细胞生长和过氧化氢酶合成; 由于 ·与细胞作用的方式和效果与H2O2不同, 超氧化物歧化酶和过氧化氢酶的快速合成并不能保证细胞及时有效地清除胞内的活性氧, 因此, 细胞对 ·胁迫的响应程度要低于H2O2胁迫; 在所考察的3种活性氧中, OH·胁迫(Fenton反应体系)对细胞的氧化损伤程度最大, 胁迫强烈地抑制了细胞生长和抗氧化物酶的合成。由此表明, 由于不同活性氧的化学性质有所不同, 细胞对不同种类、程度和形式的活性氧胁迫会表现出不同的生物学效应, 为了提高自身对氧胁迫的抵抗能力, 微生物会通过自身的代谢调节适应新的环境, 包括调整抗氧化物酶合成水平、改变生长速度以及底物消耗速率等。     

不同活性氧胁迫下Bacillus sp. F26以抗氧化物酶合成为特征的应激响应

采用不同的活性氧发生源, 研究了· 、H2O2和OH·胁迫下Bacillus sp. F26以抗氧化物酶合成为特征的应激响应。结果表明, 细胞对氧胁迫的应激响应程度取决于活性氧种类、胁迫程度和形式(瞬时和持续)。Bacillus sp. F26对H2O2胁迫的响应程度最高, 过氧化氢酶的快速合成对细胞抵抗H2O2胁迫至关重要, 当细胞及时分解进入胞内的H2O2, 胁迫对细胞的氧化损伤程度并不高, 相反会刺激细胞的生长和底物消耗, 当胁迫超过过氧化氢酶的分解能力时, H2O2会迅速抑制细胞生长和过氧化氢酶合成; 由于 ·与细胞作用的方式和效果与H2O2不同, 超氧化物歧化酶和过氧化氢酶的快速合成并不能保证细胞及时有效地清除胞内的活性氧, 因此, 细胞对 ·胁迫的响应程度要低于H2O2胁迫; 在所考察的3种活性氧中, OH·胁迫(Fenton反应体系)对细胞的氧化损伤程度最大, 胁迫强烈地抑制了细胞生长和抗氧化物酶的合成。由此表明, 由于不同活性氧的化学性质有所不同, 细胞对不同种类、程度和形式的活性氧胁迫会表现出不同的生物学效应, 为了提高自身对氧胁迫的抵抗能力, 微生物会通过自身的代谢调节适应新的环境, 包括调整抗氧化物酶合成水平、改变生长速度以及底物消耗速率等。     

盐胁迫过程中抗坏血酸对植物的保护功能

以拟南芥抗坏血酸突变体 vtc- 1 和野生型 wt 为材料 ,研究了抗氧化系统对盐胁迫的响应机制 ,以揭示抗坏血酸 ASA 的抗氧化机理及对植物的保护功能 .结果显示 :10 0 mm ol/ L Na Cl处理 12、2 4、4 8、72 h,vtc- 1和 wt体内 MDA 丙二醛 及 H2 O2 过氧化氢 的含量均明显增加 ,但 vtc- 1增加的程度明显高于 wt,说明盐胁迫可能对vtc- 1造成了更严重的氧化伤害 .胁迫过程中 ,wt体内的几种抗氧化酶 [超氧化物歧化酶 SOD 、过氧化氢酶 CAT 、抗坏血酸过氧化物酶 APX ]活性均升高 ,而 vtc- 1体内 SOD、CAT活性降低 ,APX活性在胁迫 2 4 h之前增加 ,2 4 h之后降低 ;同时 ,vtc- 1中总的抗坏血酸含量和还原型谷胱甘肽 /氧化型谷胱甘肽 GSH/ GSSG 的比值下降 ,而 wt与此相反 .本研究表明 :抗坏血酸参与活性氧 AOS 的代谢 ,减轻 AOS对植物的伤害 ;并可能对植物细胞内的抗氧化酶具有调节作用 ,增强逆境胁迫下植物的抗逆能力 ,对植物有重要的生物学保护功能     

Proline and glycinebetaine enhance antioxidant defense and methylglyoxal detoxification systems and reduce NaCl-induced damage in cultured tobacco cells

Salt stress impairs reactive oxygen species (ROS) and methylglyoxal (MG) detoxification systems, and causes oxidative damage to plants. Up-regulation of the antioxidant and glyoxalase systems provides protection against NaCl-induced oxidative damage in plants. Thiol–disulfide contents, glutathione content and its associated enzyme activities involved in the antioxidant defense and glyoxalase systems, and protein carbonylation in tobacco Bright Yellow-2 cells grown in suspension culture were investigated to assess the protection offered by proline and glycinebetaine against salt stress. Salt stress increased protein carbonylation, contents of thiol, disulfide, reduced (GSH) and oxidized (GSSG) forms of glutathione, and the activity of glutathione-S-transferase and glyoxalase II enzymes, but decreased redox state of both thiol–disulfide and glutathione, and the activity of glutathione peroxidase and glyoxalase I enzymes involved in the ROS and MG detoxification systems. Exogenous application of proline or glycinebetaine resulted in a reduction of protein carbonylation, and in an increase in glutathione redox state and activity of glutathione peroxidase, glutathione-S-transferase and glyoxalase I under salt stress. Neither proline nor glycinebetaine, however, had any direct protective effect on NaCl-induced GSH-associated enzyme activities. The present study, therefore, suggests that both proline and glycinebetaine provide a protective action against NaCl-induced oxidative damage by reducing protein carbonylation, and enhancing antioxidant defense and MG detoxification systems.     

Susceptibility to oxidative stress in Adélie and emperor penguin

The antioxidant defences in aerobic organisms represent the detoxification pathway against toxicity of reactive oxygen species (ROS). These highly reactive molecules are normally produced during the 4-electrons reduction of molecular oxygen to water coupled with oxidative phosphorylation, and during the activity of several enzymatic systems which produce ROS as intermediates. However, the endogenous generation of oxyradicals may be influenced by different environmental and biological factors, and the basal efficiency of antioxidant systems generally reflects the normal prooxidant pressure to which organisms are exposed. If the antioxidant capacity is exceeded (i.e. as a consequence of enhanced intracellular formation of ROS), a pathological condition, generally termed oxidative stress, may arise. In this preliminary work, susceptibility to oxidative stress has been compared in plasma of Adélie penguin (Pygoscelis adeliae), emperor penguin (Aptenodytes forsteri), south polar skua (Catharacta maccormicki) and snow petrel (Pagodroma nivea). Within the framework of the Italian Research Program in Antarctica, blood samples were collected during the austral summer 1998-1999 and the Total Oxyradical Scavenging Capacity (TOSC) analysed. The TOSC assay, measuring the capability of biological samples to neutralise different oxyradicals, has been recently standardised to provide a quantifiable value of biological resistance to toxicity of ROS. Penguins exhibited higher scavenging capacity towards peroxyl radicals than south polar skua and snow petrel. The greater resistance to toxicity of oxyradicals might suggest that penguins are naturally exposed to a higher basal prooxidant pressure in comparison to other analysed Antarctic birds.     

Multifunctional role of a fungal pathogen-secreted laccase 2 in evasion of insect immune defense

Laccases are widely present in bacteria, fungi, plants and invertebrates and involved in a variety of physiological functions. Here, we report that Beauveria bassiana, an economic important entomopathogenic fungus, secretes a laccase 2 (BbLac2) during infection that detoxifies insect immune response-generated reactive oxygen species (ROS) and interferes with host immune phenoloxidase (PO) activation. BbLac2 is expressed in fungal cells during proliferation in the insect haemocoel and can be found to distribute on the surface of haemolymph-derived in vivo fungal hyphal bodies or be secreted. Targeted gene-knockout of BbLac2 increased fungal sensitivity to oxidative stress, decreased virulence to insect, and increased host PO activity. Strains overexpressing BbLac2 showed increased virulence, with reduced host PO activity and lowered ROS levels in infected insects. In vitro assays revealed that BbLac2 could eliminate ROS and oxidize PO substrates (phenols), verifying the enzymatic functioning of the protein in detoxification of cytotoxic ROS and interference with the PO cascade. Moreover, BbLac2 acted as a cell surface protein that masked pathogen associated molecular patterns (PAMPs), enabling the pathogen to evade immune recognition. Our data suggest a multifunctional role for fungal pathogen-secreted laccase 2 in evasion of insect immune defenses.     

In vivo ROS production and use of oxidative stress-derived biomarkers to detect the onset of diseases such as Alzheimer’s disease,Parkinson’s disease,and diabetes

Breakthroughs in biochemistry have furthered our understanding of the onset and progression of various diseases, and have advanced the development of new therapeutics. Oxidative stress and reactive oxygen species (ROS) are ubiquitous in biological systems. ROS can be formed non-enzymatically by chemical, photochemical and electron transfer reactions, or as the byproducts of endogenous enzymatic reactions, phagocytosis, and inflammation. Imbalances in ROS homeostasis, caused by impairments in antioxidant enzymes or non-enzymatic antioxidant networks, increase oxidative stress, leading to the deleterious oxidation and chemical modification of biomacromolecules such as lipids, DNA, and proteins. While many ROS are intracellular signaling messengers and most products of oxidative metabolisms are beneficial for normal cellular function, the elevation of ROS levels by light, hyperglycemia, peroxisomes, and certain enzymes causes oxidative stress-sensitive signaling, toxicity, oncogenesis, neurodegenerative diseases, and diabetes. Although the underlying mechanisms of these diseases are manifold, oxidative stress caused by ROS is a major contributing factor in their onset. This review summarizes the relationship between ROS and oxidative stress, with special reference to recent advancements in the detection of biomarkers related to oxidative stress. Further, we will introduce biomarkers for the early detection of neurodegenerative diseases and diabetes, with a focus on our recent work.     

The role of protein quality control in mitochondrial protein homeostasis under oxidative stress

Mitochondria contribute significantly to the cellular production of ROS. The deleterious effects of increased ROS levels have been implicated in a wide variety of pathological reactions. Apart from a direct detoxification of ROS molecules, protein quality control mechanisms are thought to protect protein functions in the presence of elevated ROS levels. The reactivities of molecular chaperones and proteases remove damaged polypeptides, maintaining enzyme activities, thereby contributing to cellular survival both under normal and stress conditions. We characterized the impact of oxidative stress on mitochondrial protein homeostasis by performing a proteomic analysis of isolated yeast mitochondria, determining the changes in protein abundance after ROS treatments. We identified a set of mitochondrial proteins as substrates of ROS‐dependent proteolysis. Enzymes containing oxidation‐sensitive prosthetic groups like iron/sulfur clusters represented major targets of stress‐dependent degradation. We found that several proteins involved in ROS detoxification were also affected. We identified the ATP‐dependent protease Pim1/LON as a major factor in the degradation of ROS‐modified soluble polypeptides localized in the matrix compartment. As Pim1/LON expression was induced significantly under ROS treatment, we propose that this protease system performs a crucial protective function under oxidative stress conditions.     

Antioxidative response mechanisms in halophytes: Their role in stress defence

Normal growth and development of plants is greatly dependent on the capacity to overcome environmental stresses. Environmental stress conditions like high salinity, drought, high incident light and low or high temperature cause major crop losses worldwide. A common denominator in all these adverse conditions is the production of reactive oxygen species (ROS) within different cellular compartments of the plant cell. Plants have developed robust mechanisms including enzymatic or nonenzymatic scavenging pathways to counter the deleterious effects of ROS production. There are a number of general reviews on oxidative stress in plants and few on the role of ROS scavengers during stress conditions. Here we review the regulation of antioxidant enzymes during salt stress in halophytes, especially mangroves. We conclude that (i) antioxidant enzymes protect halophytes from deleterious ROS production during salt stress, and (ii) genetic information from mangroves and other halophytes would be helpful in defining the roles of individual isoforms. This information would be critical in using the appropriate genes for oxidative stress defence for genetic engineering of enhanced stress tolerance in crop systems.     

Human SOD2 modification by dopamine quinones affects enzymatic activity by promoting its aggregation: possible implications for Parkinson's disease

Mitochondrial dysfunction and oxidative stress are considered central in dopaminergic neurodegeneration in Parkinson's disease (PD). Oxidative stress occurs when the endogenous antioxidant systems are overcome by the generation of reactive oxygen species (ROS). A plausible source of oxidative stress, which could account for the selective degeneration of dopaminergic neurons, is the redox chemistry of dopamine (DA) and leads to the formation of ROS and reactive dopamine-quinones (DAQs). Superoxide dismutase 2 (SOD2) is a mitochondrial enzyme that converts superoxide radicals to molecular oxygen and hydrogen peroxide, providing a first line of defense against ROS. We investigated the possible interplay between DA and SOD2 in the pathogenesis of PD using enzymatic essays, site-specific mutagenesis, and optical and high-field-cw-EPR spectroscopies. Using radioactive DA, we demonstrated that SOD2 is a target of DAQs. Exposure to micromolar DAQ concentrations induces a loss of up to 50% of SOD2 enzymatic activity in a dose-dependent manner, which is correlated to the concomitant formation of protein aggregates, while the coordination geometry of the active site appears unaffected by DAQ modifications. Our findings support a model in which DAQ-mediated SOD2 inactivation increases mitochondrial ROS production, suggesting a link between oxidative stress and mitochondrial dysfunction.     

Antioxidant responses of wheat plants under stress

Currently, food security depends on the increased production of cereals such as wheat (Triticum aestivum L.), which is an important source of calories and protein for humans. However, cells of the crop have suffered from the accumulation of reactive oxygen species (ROS), which can cause severe oxidative damage to the plants, due to environmental stresses. ROS are toxic molecules found in various subcellular compartments. The equilibrium between the production and detoxification of ROS is sustained by enzymatic and nonenzymatic antioxidants. In the present review, we offer a brief summary of antioxidant defense and hydrogen peroxide (H₂O₂) signaling in wheat plants. Wheat plants increase antioxidant defense mechanisms under abiotic stresses, such as drought, cold, heat, salinity and UV-B radiation, to alleviate oxidative damage. Moreover, H₂O₂ signaling is an important factor contributing to stress tolerance in cereals.     

The roles of cellular reactive oxygen species,oxidative stress and antioxidants in pregnancy outcomes

Reactive oxygen species (ROS) are generated as by-products of aerobic respiration and metabolism. Mammalian cells have evolved a variety of enzymatic mechanisms to control ROS production, one of the central elements in signal transduction pathways involved in cell proliferation, differentiation and apoptosis. Antioxidants also ensure defenses against ROS-induced damage to lipids, proteins and DNA. ROS and antioxidants have been implicated in the regulation of reproductive processes in both animal and human, such as cyclic luteal and endometrial changes, follicular development, ovulation, fertilization, embryogenesis, embryonic implantation, and placental differentiation and growth. In contrast, imbalances between ROS production and antioxidant systems induce oxidative stress that negatively impacts reproductive processes. High levels of ROS during embryonic, fetal and placental development are a feature of pregnancy. Consequently, oxidative stress has emerged as a likely promoter of several pregnancy-related disorders, such as spontaneous abortions, embryopathies, preeclampsia, fetal growth restriction, preterm labor and low birth weight. Nutritional and environmental factors may contribute to such adverse pregnancy outcomes and increase the susceptibility of offspring to disease. This occurs, at least in part, via impairment of the antioxidant defense systems and enhancement of ROS generation which alters cellular signalling and/or damage cellular macromolecules. The links between oxidative stress, the female reproductive system and development of adverse pregnancy outcomes, constitute important issues in human and animal reproductive medicine. This review summarizes the role of ROS in female reproductive processes and the state of knowledge on the association between ROS, oxidative stress, antioxidants and pregnancy outcomes in different mammalian species.     

A comparison of arteries and veins in oxidative stress: producers, destroyers, function, and disease

Reactive oxygen species (ROS) are by-products of oxygen metabolism, normally present in low levels inside cells, where they participate in signaling processes. The delicate balance in the continuous cycle of ROS generation and inactivation is maintained by enzymatic and nonenzymatic endogenous systems. Overwhelming production of ROS (by such sources as the mitochondrial electron transport chain, NADPH oxidase, xanthine oxidase, or uncoupled nitric oxide synthase), when inadequately counteracted by destruction through antioxidant systems (such as superoxide dismutase or catalase), leads to a prooxidant state also known as oxidative stress. Increased levels of ROS and markers of oxidative stress have been consistently found in such cardiovascular diseases as atherosclerosis or hypertension, although controversy still exists over the pathophysiological role of oxidative stress in these conditions. ROS can modulate vascular function either by direct oxidative damage or by activating cellular signaling pathways that lead to abnormal contractile, inflammatory, proliferative, or remodeling properties of the blood vessel. Most current research focuses on these processes in arteries, leaving veins, "the other side" of vascular biology, in obscurity. Veins are different structurally and functionally from arteries. Equipped with a smaller smooth muscle layer compared to arteries, but being able to accommodate 70% of the circulating blood volume, veins can modulate cardiovascular homeostasis and contribute significantly to hypertension pathogenesis. Although the reports on the quantitative differences in ROS production in veins compared to arteries had conflicting results, there is a clear qualitative difference in ROS metabolism and utilization between the two vessel types. This review will compare and contrast the current knowledge of ROS metabolism in arteries versus veins in both physiological and pathophysiological conditions. Our understanding of the mechanisms underlying vascular diseases would greatly benefit from a more thorough exploration of the role of veins and venous oxidative stress.