超氧化物歧化酶

本文就超氧化物歧化酶的种类、分布、结构、活性部位、催化机理、纯化、活力测定、理化特性等方面作了介绍,并叙述了它的研究动向和发展前景。     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

NaCl胁迫下枸杞愈伤组织活性氧产生与质膜H+—ATPase活性的关系

以枸杞愈伤组织为材料,研究了盐胁迫对活性氧伤害和质膜H -ATPase活性的影响。结果表明,NaCl浓度为100mmol/L时超氧阴离子(O2-·)和过氧化氢(H2O2)含量上升,质膜H -ATPase活性先升后降;质膜相对透性与丙二醛(MDA)含量呈显著正相关;超氧阴离子(O2-·)和过氧化氢(H2O2)含量与质膜H -ATPase活性在重度胁迫下呈显著负相关,说明盐胁迫下活性氧积累可能是加速伤害质膜功能的主要原因之一。     

Differential levels of antioxidants in paraquat-resistant and -susceptible Erigeron canadensis biotypes in Korea

To characterize the biochemical differences in paraquat-resistant and -susceptible biotypes of Erigeron canadensis L. collected from Korea, we investigated the constitutive levels of various antioxidants such as antioxidant enzymes and low molecular weight antioxidants in leaves, as well as after paraquat treatment. The activities of superoxide dismutase, peroxidase, ascorbate peroxidase, and catalase were higher in the paraquat-resistant biotype than in the paraquat-susceptible biotype. Reduced ascorbic acid content was higher in the resistant biotype, but the content of reduced glutathione was higher in the susceptible biotype. These results indicate that one of the paraquat-resistant mechanisms in E. canadensis in the present study might be related to protecting the activities of antioxidant enzymes, such as superoxide dismutase, peroxidase ascorbate peroxidase, and catalase, as well as the contents of low molecular weight antioxidants such as ascorbate and glutathione.     

Footsteps from Insect Larvae Damage Leaf Surfaces and Initiate Rapid Responses

Plant resistance to insect herbivory involves gene expression in response to wounding and the detection of insect elicitors in oral secretions (Kessler and Baldwin, 2002, Ann. Rev. Plant/ Biol. 53: 299–328). However, crawling insect larvae stimulate the synthesis of 4-aminobutyrate within minutes and imprints of larval footsteps can be visualized within seconds through superoxide production or transient increases in chlorophyll fluorescence (Bown et al., 2002, Plant Physiol. 129: 1430–1434). Here cryo-scanning electron microscopy was used to demonstrate that larval feet, which are equipped with a perimeter row of hook-like crochets, damage leaf tissue and result in larval footprints. Staining for cell death shows that areas of wounding correspond to footsteps detected through increased chlorophyll fluorescence. Superoxide production in response to footsteps was inhibited by diphenyleneiodonium, an inhibitor of the plasma membrane NADPH oxidase enzyme. Inhibition of superoxide production, however, did not eliminate the detection of cell death. The results demonstrate that larval footsteps damage leaf tissue, and initiate rapid local responses which are not dependent on herbivory or oral secretions. It is proposed that superoxide production at the wound site prevents opportunistic pathogen infection.     

蝴蝶兰切花花被不同开放阶段酶活性的变化

以蝴蝶兰鲜切花为试材,对切花花被不同生理阶段3种酶活性的变化进行测量分析,以期为蝴蝶兰切花寿命的延长提供参考依据。结果表明:萼片和花瓣中的过氧化物酶(POD)从花苞期到半开期酶活性变化不明显,从半开期到衰老期酶活性显著升高。花瓣中的超氧化物歧化酶(SOD)从花苞期到半开期酶活性升高,从半开期到衰老期酶活性显著降低;萼片中的超氧化物歧化酶活性变化不显著但也呈先上升后下降趋势。花瓣中的中性蛋白酶(NP)从花苞期到半开期酶活性呈上升趋势,半开期到盛开期呈下降趋势,盛开期到衰老期呈上升趋势;花萼中的中性蛋白酶从花苞期到盛开期酶活性呈下降趋势,从盛开期到衰老期呈上升趋势;总体来看中性蛋白酶的活性呈先下降后上升趋势。     

超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响

试验旨在探究在H₂O₂诱导的氧化应激状态下，超氧化物歧化酶模拟物（SODm）对仔猪空肠上皮细胞系（IPEC-J2）的保护作用。利用MTT法筛选出构建氧化应激模型H₂O₂的适宜浓度；将IPEC-J2细胞分别用0、0.05、0.5、5、50、500、2 000 U/mL SODm进行培养，利用MTT法分别在2、4、8、12 h时测定各组细胞存活率，筛选出SODm作用的适宜浓度和时间；根据构建的氧化应激模型和SODm适宜浓度和时间，将IPEC-J2细胞随机分为空白组、模型组、SODm处理组（SODm_0.5、SODm₅、SODm₅₀）和超氧化物歧化酶（SOD）处理组（SOD_0.5、SOD₅、SOD₅₀），分别测定各组细胞存活率、活性氧（ROS）含量及细胞内SOD、谷胱甘肽过氧化物酶（GSH-Px）活性及丙二醛（MDA）含量和总抗氧化能力（T-AOC）。结果表明：①H₂O₂构建细胞氧化应激模型的适宜浓度是1.0 mmol/mL。②当不同浓度SODm分别作用4、8、12 h时，0.5~50 U/mL SODm组细胞存活率显著高于空白组（P<0.05）；且8 h时存活率最高，在12 h时处于下降趋势。③除空白组外，SODm₅和SOD₅预处理的IPEC-J2存活率显著高于其他组（P<0.05）；且SODm和SOD组中细胞ROS含量显著低于模型组（P<0.05）；模型组与空白组相比，均显著降低了SOD、GSH-Px活性和T-AOC水平，提高了MDA含量（P<0.05）；与模型组相比，所有SODm和SOD处理组均显著提高了SOD、GSH-Px活性和T-AOC水平，降低了MDA含量（P<0.05），同时SODm₅₀组T-AOC水平显著低于SOD₅₀组（P<0.05）。综上，SODm对H₂O₂诱导氧化损伤状态下IPEC-J2具有保护作用。     

Novel Caryophyllane-Related Sesquiterpenoids with Anti-Inflammatory Activity from Rumphella antipathes (Linnaeus, 1758)

Two previously undescribed caryophyllane-related sesquiterpenoids, antipacids A (1) and B (2), with a novel bicyclo[5.2.0] core skeleton, and known compound clovane-2β,9α-diol (3), along with rumphellolide L (4), an esterified product of 1 and 3, were isolated from the organic extract of octocoral Rumphella antipathes. Their structures, including the absolute configurations were elucidated by spectroscopic and chemical experiments. In vivo anti-inflammatory activity analysis indicated that antipacid B (2) inhibited the generation of superoxide anions and the release of elastase by human neutrophils, with IC₅₀ values of 11.22 and 23.53 μM, respectively, while rumphellolide L (4) suppressed the release of elastase with an IC₅₀ value of 7.63 μM.     

红素氧还蛋白研究进展

自然界中大多数的红素氧还蛋白(rubredoxin)集中来自严格厌氧菌,包括细菌、古菌和少数微需氧细菌,而植物中该类蛋白相对较少,它的功能主要表现在作为辅因子或电子供体参与多个生化途径。也有研究显示,无论是植物中还是微生物中的红素氧还蛋白基因在协助生物抵御氧化胁迫方面均发挥着重要的作用。本文全面介绍自然界中的红素氧还蛋白基因的分布、结构以及在微生物、植物中所发挥的作用等,希望能为今后的进一步研究提供参考。     

Effects of cerium oxide supplementation to laying hen diets on performance,egg quality,some antioxidant enzymes in serum and lipid oxidation in egg yolk

This study was conducted to determine the effects of dietary cerium oxide levels (0, 100, 200, 300 or 400 mg/kg) on the laying performance, egg quality, some blood serum parameters and egg lipid peroxidation of laying hen. In total, one hundred and twenty 22‐week‐old brown Lohman LSL laying hens were randomly assigned to five groups equally (n = 24). Each treatment was replicated six times. Dietary supplementation of cerium oxide had no significant effect on feed intake and egg weight. The addition of cerium oxide to the laying hens' feed improved feed conversion ratio and increased (p < 0.05) egg production. Quality criteria of egg for except shell breaking strength were not affected by supplementing cerium oxide. In particular, supplementation of 200 and 300 mg/kg cerium oxide to the laying hens feed led to a significant (p < 0. 01) increase in egg shell breaking strength. Calcium and phosphorus concentration of serum increased significantly (p < 0.05) with supplementation of 100 mg/kg cerium oxide to laying hen diets. It was also observed that serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration decreased significantly with supplementation of cerium oxide in diets. Inclusion of cerium oxide resulted in a significant reduction in thiobarbituric acid reactive substance (TBARS) values in egg yolk in this study. It can be concluded that the addition of cerium oxide had positive effects on egg production, feed conversion ratio and egg shelf life. Based on the results of this study, it could be advised to supplement laying hens feed with cerium oxide as feed additives.