淡水三角涡虫不同发育时期三种抗氧化酶的研究

以鲁中山地区的淡水三角涡虫卵囊、幼虫、成虫为材料,研究了涡虫在不同发育过程中3种抗氧化酶SOD、CAT、GSH-Px的活性变化.结果 表明,SOD在发育初期活性增长迅速,在幼虫孵出后活性略减,最后趋于稳定;CAT活性在卵囊阶段活性较低,从幼虫孵出后活性增长很快,并在成体中保持较高的活性;GSH-Px活性在卵囊时期活性较高,从幼虫孵出后活性降低,在成体中活性较低.     

Antioxidant enzymes of the black swallowtail butterfly,Papilio polyxenes,and their response to the prooxidant allelochemical,quercetin

The black swallowtail butterfly larvae, Papilio polyxenes, are specialist feeders that have adapted to feeding on plants containing high levels of prooxidant allelochemicals. Third, fourth, and fifth instar larvae were tested for their antioxidant enzyme activities, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPOX), using 850-g supernatants from whole-body homogenates. The overall antioxidant enzyme profile for P. polyxenes was high compared to other insects, with activities ranging as follows: SOD, 1.1–7.5; CAT, 124–343; GR, 1.0–7.5; and GPOX, 0 units. To determine whether these antioxidant enzymes were inducible, P. poly xenes larvae were given a prooxidant challenge by dipping parsley leaves (their diet in the initial studies) in solutions of quercetin, such that the leaves became coated with this prooxidant flavonoid. Mid-fifth instar larvae fed on quercetin-coated leaves were assayed for antioxidant enzyme activities as was previously done with the larvae fed the standard diet. Food consumption and quercetin intake were monitored. SOD activity was increased almost twofold at the highest quercetin concentration tested. CAT and GR activity, on the other hand, were inhibited by increased quercetin consumption, with GR activity completely inhibited at the highest quercetin concentration after 12 h of feeding. GPOX activity, not present in control insects, was also not inducible by a quercetin challenge. These studies point out the key role that the antioxidant enzymes play in insect defenses against plant prooxidants.     

Antioxidant enzyme activities in subcellular fractions of larvae of the black swallowtail butterfly,Papilio polyxenes

The black swallowtail butterfly, Papilio polyxenes, larvae are specialized feeders of pro-oxidant rich plants of Apiaceae and Rutaceae. An important defense against toxic forms of oxygen species generated by ingestion of the pro-oxidants, are the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), GSH-dependent glutathione peroxidases (selenium-dependent glutathione peroxidase [GPOX] and peroxidase activity of selenium-independent glutathione-S-transferase [GT_px]), and glutathione reductase (GR). The subcellular distribution of these enzymes in black swallowtail larvae was investigated and was found to resemble the patterns described for larvae of two other lepidopteran species: the southern armyworm, Spodoptera eridania, and the cabbage looper, Trichoplusia ni. The confinement of SOD in the cytosol and mitochondria was typically eukaryotic, but the relative proportion (1:1) was markedly different from the mammalian pattern (4:1; cytosol:mitochondria). The most obvious difference between the black swallowtail and other lepidoptera as a group, and mammalian species, is in very wide intracellular distributions of CAT, GTpx, and GR in insect species. Insects possess very low levels of a GPOX-like activity which reduces both H₂O₂ and organic peroxides. Consequently, insects have elaborate activities with a wide subcellular distribution of both CAT which decomposes H₂O₂, and GTpx which decomposes organic peroxides. The reduction of peroxides is dependent on GSH, which in this process is oxidized to GSSG. GR which reduces GSSG to GSH is also of wide subcellular distribution, analogous to the distribution pattern of GTpx.     

Antioxidant enzymes as biochemical defenses against phototoxin-induced oxidative stress in three species of herbivorous lepidoptera

Many secondary plant compounds are capable of photoactivation resulting in the production of toxic species of oxygen. One mechanism of defense for insects feeding on phototoxic plants may be the presence of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR). The activities of these enzymes were examined in larvae of three lepidoptera: Ostrinia nubilalis, Manduca sexta, and Anaitis plagiata. Highest levels of antioxidant enzyme activity were found in A. plagiata, a specialist feeder on Hypericum perforatum, which contains high levels of the phototoxin hypericin. Larvae of A. plagiata fed leaf discs treated with hypericin exhibited a short-term, concentration-dependent decline in enzyme activity. Longer term studies with A. palgiata fed either the photoxic H. perforatum, or the closely related but non-phototoxic H. calycinum, resulted in increased CAT and GR activity in larvae fed the phototoxic plant whereas SOD activity was not significantly different. These results suggest that CAT and GR may be inducible defenses against phototoxins.     

Activity Development of Selected Detoxication Enzymes during the Ontogenesis of the Zebrafish (Danio rerio)

Activities of two detoxication enzyme systems, microsomal and soluble glutathione S‐transferases (m‐/sGST, EC 2.5.1.18), and Se‐dependent glutathione peroxidase (GP‐X, EC 1.11.1.9) were monitored during zebrafish (Danio rerio) development. Starting with the 2–4 cell stage of ontogenesis successively up to adult fish, the presence of these detoxication enzymes could be demonstrated. Activities increased slowly during development and showed elevations corresponding to increased stress conditions. Plain increase occured after hatching, possibly caused by the change of the surrounding micromilieu. The juveniles showed highest activities, certainly provoked by new substances as from the food. Enzyme activities of adults revealed moderate levels probably due to a dilution of highly concentrated detoxication activities in the ready developed organs, especially the liver, in the whole body homogenate.     

Activities of enzymes that detoxify superoxide anion and related toxic oxyradicals in Trichoplusia ni

In third-, fourth-, and fifth-instar larvae of the cabbage looper moth, Trichoplusia ni, the activities of the antioxidant enzymes, superoxide dismutase (SOD*), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR) were examined using 850 g supernatants of whole-body homogenates. The enzyme activities, expressed as units mg⁻¹ protein min⁻¹ at 25°C ranged as follows: SOD, 0.67-2.13 units; CAT, 180.5-307.5 units; GPOX, none detectable; and GR, 0.40-1.19 units. There was a similar pattern of changes for SOD and CAT activities with larval ontogeny, but not for GR. The cabbage looper apparently uses SOD and CAT to form a “defensive team” effective against endogenously produced superoxide anion (O₂⪸). Glutathione may serve as an antioxidant for the destruction of any organic/lipid peroxides formed, and GSH oxidized to glutathione disulfide would be recycled by GR. Bioassays against pro-oxidant compounds exogenous sources of (O₂⪸) show high sensitivity of mid-fifth instars to the linear furanocoumarin, 8-methoxypsoralen (xanthotoxin) primarily from photoactivation (320-380 nm), and auto-oxidation of the flavonoid, quercetin. The LC₅₀s are 0.0004 and 0.0045% (w/w) concentration of xanthotoxin and quercetin, respectively. Both pro-oxidants have multiple target sites for lethal action and, in this context, the role of antioxidant enzymes is discussed.     

Changes in the Activity of Antioxidant Enzymes in Wheat Leaves and Roots as a Function of Nitrogen Source and Supply

The activity of enzymes participating in the systems of antioxidant protection was assayed in the second leaf and roots of 21-day-old wheat seedlings (Triticum aestivum L.) grown in a medium with nitrate (NO^– ₃ treatment), ammonium (NH⁺ ₄ treatment), or without nitrogen added (N-deficiency treatment). The activities of superoxide dismutase (SOD), peroxidase, ascorbate peroxidase, glutathione reductase, and catalase in the leaves and roots of the NH⁺ ₄ plants was significantly higher than in the plants grown in the nitrate medium. The activity of SOD decreased and ascorbate peroxidase markedly increased in leaves, whereas the activity of ascorbate peroxidase increased in the roots of N-deficient plants, as compared to the plants grown in nitrate and ammonium. Low-temperature incubation (5°, 12 h) differentially affected the antioxidant activity of the studied plants. Whereas leaf enzyme activities did not change in the NH⁺ ₄ plants, the activities of SOD, peroxidase, ascorbate peroxidase, and catalase markedly increased in the NO^– ₃ plants. In leaves of the N-deficient plant, the activity of SOD decreased; however, the activity of other enzymes increased. In response to temperature decrease, catalase activity increased in the roots of NO^– ₃ and NH⁺ ₄-plants, whereas in the N-deficient plants, the activity of peroxidase increased. Thus, in wheat, both nitrogen form and nitrogen deficiency changed the time-course of antioxidant enzyme activities in response to low temperature.     

Action of antioxidant enzymes and cytochrome P-450 monooxygenases in the cabbage looper in response to plant phototoxins

Effects of two biosynthetically distinct plant phototoxins—xanthototoxin, a furanocoumarin, and harmine, a β-carboline alkaloid, which are known to produce toxic oxygen species—on the food utilization efficiencies and enzymatic detoxification systems of the polyphagous cabbage looper. Trichoplusia ni (Lepidoptera: Noctuidae), were studied. Newly molted fifth-instar larvae were allowed 36 h to ingest diets containing these two phototoxins at 0.15% wet weight in the presence of near ultraviolet (UVA). The growth and development of the larvae, as well as the corresponding activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR) and the detoxification enzyme cytochrome P-450, were measured. Xanthotoxin reduced rates of relative growth and consumption and efficiencies of conversion of ingested and digested food to biomass. Harmine reduced rates of growth and consumption without affecting efficiencies of conversion. Specific activities of SOD, CAT, GPOX, and GR of whole-body homogenates in the absence of compounds were 0.88 units, 153μmol H₂O₂ decomposed·mg protein^?1·min—¹, 38.3 nmol NADPH oxidized·mg protein^?1·min^?1, and 0.56 nmol NADPH oxidized·mg protein^?1·min^?1, respectively. SOD activity was induced 2.9-fold and 3.8-fold by dietary xanthotoxin and harmine, respectively. CAT and GPOX activities were induced 1.2-fold by harmine only, and GR activity was not changed by either chemical. The P-450 activity toward xanthotoxin in the microsomal fraction of midguts was low (0.15 nmol xanthotoxin metabolized·mg protein^?1·min^?1) and was not induced by xanthotoxin ingestion. These studies indicate that P-450 and antioxidant enzyme systems may be independent but consequential, the induction of antioxidant enzymes by phototoxins occurring when low P-450 activity toward the phototoxin permits the accumulation of oxidative stress from unmetabolized phototoxin, which in turn induces antioxidant enzymes.     

Therapeutic Usages of Oxygen Radical Scavengers in Human Diseases: Myths and Realities

Some distinct advances in pharmacologic manipulation of oxygen radical scavengers have been made which could ultimately greatly enhance the use of these reagents as drugs, as well as some innovative techniques for drug delivery. Unfortunately, most of the therapeutic reports in the literature, almost all of which are based on usage of standard (native) SOD and/or catalase, are still anecdotal and/or uncontrolled. A review of the human disedse/treatment literature suggests that further tightening of the scientific design of such trials is stjll badly needed; hopefully better experimental design will be applied when products such as PEG conjugates or genetically engineered polymers are ready for testing.     

Glutathione and Its Metabolic Enzymes in Gliomal Tumor Tissue and the Peritumoral Zone at Different Degrees of Anaplasia

This research was aimed at investigating the features of free radical activity and the parameters of glutathione metabolism in tumor tissues and the peritumoral zone at different degrees of glial tumor anaplasia. We analyzed postoperative material from 20 patients with gliomas of different degrees of anaplasia. The greatest differences compared to adjacent noncancerous tissues were found in the tumor tissue: an increased amount of glutathione and glutathione-related enzymes at Grades I and II, and a decrease of these parameters at Grades III and IV. For the peritumoral zone of Grades I and II, the indices changed in different directions, while for Grades III and IV, they occurred synchronously with the tumor tissue changes. For Low Grade and High Grade gliomas, opposite trends were revealed regarding changes in the level of glutathione and the enzymes involved in its metabolism and in the free radical activity in the peritumoral zone. The content of glutathione and the enzymes involved in its metabolism decreased with the increasing degree of glioma anaplasia. In contrast, free radical activity increased. The glutathione system is an active participant in the antioxidant defense of the body and can be used to characterize the cell condition of gliomas at different stages of tumor development.     

Responses of antioxidative system to chilling stress in two rice cultivars differing in sensitivity

The responses of antioxidative system of rice to chilling were investigated in a tolerant cultivar, Xiangnuo-1, and a susceptible cultivar, IR-50. The electrolyte leakage and malondialdehyde content of Xiangnuo-1 were little affected by chilling treatment but those of IR-50 increased. Activities of suoperoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase, and ascorbic acid content of Xiangnuo-1 were remained high, while those of IR-50 decreased under chilling. The results indicated that higher activities of defense enzymes and higher content of antioxidant under stress were associated with tolerance to chilling.     

Influence of a Brassinosteroid Analogue on Antioxidant Enzymes in Rice Grown in Culture Medium with NaCl

We studied the effects of a polyhydroxylated spirostanic brassinosteroid analogue (BB-16) on the activities of antioxidant enzymes in rice seedlings grown in vitro in culture medium supplemented with NaCl. Seedlings were grown in medium with 75 mM NaCl and 0.001 or 0.01 mg dm⁻³ BB-16 for 16 d or 3-d-old seedlings were exposed for 4 d to 0, 0.001 or 0.01 mg dm⁻³ BB-16 then further grown in medium with 75 mM NaCl without BB-16. Seedlings exposed to 0.01 mg dm⁻³ BB-16 for 16 d showed significant increase in the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione reductase (GR) and a slight increase in ascorbate peroxidase (APX). On the other hand, 4-d exposure to BB-16 only increased SOD and CAT activities at concentration 0.001 mg dm⁻³. GR activity was not altered by this BB-16 treatment. These results indicated that BB-16, which is structurally modified in the lateral chain in relation to natural brassinosteroids, changes the activity of key antioxidant enzymes, which might confer tolerance to saline stress. This revised version was published online in July 2006 with corrections to the Cover Date.     

Subcellular distribution and activities of superoxide dismutase,catalase, glutathione peroxidase,and glutathione reductase in the southern armyworm,Spodoptera eridania

In mid-fifth-instar larvae of the southern armyworm, Spodoptera eridania, the subcellular distribution of four antioxidant enzymes—superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPOX), and glutathione reductase (GR)—were examined. Two-thirds (4.26 units ·mg protein^?1) of the SOD activity was found in the cytosol, and one-thirds (2.13 units ·mg protein^?1) in the mitochondria. CAT activity was unusually high and not restricted to the microsomal fraction where peroxisomes are usually isolated. The activity was distributed as follows: cytosol (163 units) mitochondria (125 units) and microsomes (119 units). Similar to CAT, the subcellular compartmentalization of both GPOX and GR was unusual. No activity was detected in the cytosol, but in mitochondria and microsomes, GR levels were 5.49 and 3.09 units. Although GPOX activity exhibited 14–16-fold enrichment in mitochondria and microsomes, respectively, over the 850g crude homogenate, the level was negligible (mitochondria = 1.4 × 10^?3 units; microsomes = 1.6 × 10^?3 units), indicating that this enzyme is absent. The unusual distribution of CAT has apparently evolved as an evolutionary answer to the absence of GR from the cytosol, and the lack of GPOX activity.     

Antioxidant defense in mitochondria during diapause and postdiapause development of European corn borer (Ostrinia nubilalis, Hubn.)

Antioxidant enzymes (CAT, catalase; GPx, selenium nondependent glutathione peroxidase; GST, glutathione-S-transferase; GR, glutathione reductase; DHAR, dehydroascorbate reductase) were determined in the mitochondria of diapausing and non-diapausing larvae and pupae of both diapausing and non-diapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). CAT, GST, and DHAR activity in mitochondria of diapausing larvae were reduced compared to non-diapausing larvae. Pupae of diapaused-larvae possessed lower GST, but higher DHAR activities compared to pupae of non-diapaused individuals. Comparison between larvae and pupae revealed lower GPx activity in the mitochondria of pupae. CAT activity in the mitochondria of pupae was higher compared to diapausing larvae, but lower than in non-diapausing ones. Correlation and canonical discriminant analyses revealed different antioxidant enzyme compositions for a particular stage and developmental pattern. Our results show that antioxidant enzymes have a similar role in the regulation of energetics in mitochondria as that in diapause and metamorphosis.     

Ultraviolet-B radiation (280–315 nm) invoked antioxidant defence systems in <Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp. and <Emphasis Type="Italic">Crotalaria juncea</Emphasis> L.

A crop legume Vigna unguiculata L. (Walp.) and a wild legume Crotalaria juncea L. were evaluated for their relative responses to the oxidative stress injury induced by various doses of UV-B radiation (UV-B, 280–315 nm; 0, 1.0, 1.4, 4.7, and 6.0 kJ m⁻² d⁻¹). A dose-dependent damage in lipid peroxidation was determined as an index of membrane injury caused by UV-B. The impact was significantly higher in V. unguiculata than in C. juncea. The specific activities of superoxide dismutase, ascorbate peroxidase, monodehydroascorbate reductase, and dehydroascorbate reductase increased directly proportional to UV-B doses. However, the activities of these enzymes were significantly higher in V. unguiculata than in C. juncea indicating that V. unguiculata was inflicted with more severe oxidative stress injury under UV-B. In C. juncea the glutathione reductase and ascorbate oxidase activities were 35 and 40 % greater than in V. unguiculata, respectively. Further, the non-enzymatic antioxidants ascorbate and glutathione, and their reduced/oxidizes ratios in C. juncea were much greater than V. unguiculata indicating C. juncea has an inherently greater antioxidative potential than V. unguiculata. Thus C. juncea is better adapted to oxidative stress than V. unguiculata by means of efficient cellular antioxidant mechanisms helping to combat the photooxidative stress injury elicited by UV-B.     

Salt Stress Injury Induces Oxidative Alterations and Antioxidative Defence in the Roots of Lemna minor

Lemna minor L. roots were treated with different concentrations of NaCl. Lipid peroxidation was investigated histochemically and biochemically. At higher NaCl concentrations an increase in staining was observed in the root apices as compared to control for lipid peroxidation and loss of membrane integrity as well as an increase in contents of thiobarbituric acid reactive substance and peroxide. Both the non-enzymic antioxidants, ascorbate and glutathione increased with the NaCl concentration in the roots. Whereas an increase in superoxide dismutase, guaiacol peroxidase, and glutathione reductase activities were marked, catalase activity decreased in the roots under NaCl stress. This revised version was published online in July 2006 with corrections to the Cover Date.     

白内障晶体中某些微量元素、与谷胱甘肽相关酶类活性的动态观察及其相互关系

本文动态观察了用平阳霉素诱发的大鼠白内障晶体中与谷胱甘肽代谢相关酶类活性和微量元素水平的变化,并与正常晶体进行比较,同时就酶活性与微量元素水平的相关性进行了检验。结果表明：（１）注射平阳霉素早期酶活性增高,谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）、谷胱甘肽还原酶（ＧＳＳＧ－Ｒ）及超氧化物歧化酶（ＳＯＤ）等活性的升高达显著水平,后期酶活性均下降,尤以ＧＳＨ－Ｐｘ、ＧＳＳＧ－Ｒ和谷胱甘肽硫转移酶（ＧＳＨ－Ｓ）等的活性降低明显;（２）ＧＳＨ－Ｐｘ和ＳＯＤ酶活性分别与Ｚｎ具有相关性（Ｐ＜０．０５）,这两种酶也分别与Ｓｅ具有高度相关性（Ｐ＜０．０１）,此两种元素在该类型白内障形成中可能有一定意义。     

The role of host generated free radicals in helminth infections: Nippostrongylus brasiliensis and Nematospiroides dubius compared

and 1986. The role of host generated free radicals in helminth infections: Nippostrongylus brasiliensis and Nematospiroides dubius compared. International Journal for Parasitology 16: 617–622. The possibility that free radicals are involved in the expulsion of Nippostrongylus brasiliensis from the small intestine of its mice host was explored by comparing the susceptibilities to free radicals, and levels of protective enzymes, of adult N. brasiliensis and Nematospiroides dubius, a closely related intestinal parasite of mice. Nippostrongylus brasiliensis was markedly more susceptible to in vitro free radical damage than N. dubius. The difference in susceptibility is probably related to differences in enzymatic protection against free oxygen radicals as N. dubius had roughly twice as much Superoxide dismutase, about 3 times as much catalase and about 4 times as much glutathione reductase as N. brasiliensis. This result may indicate that N. dubius persists in the rodent small intestine, whilst N. brasiliensis is spontaneously expelled, because of a more efficient enzymatic defence system against host-generated free oxygen radicals.     

Cytokinin-induced activity of antioxidant enzymes in transgenic Pssu-ipt tobacco during plant ontogeny

Cytokinin (CK) content and activities of several antioxidant enzymes were examined during plant ontogeny with the aim to elucidate their role in delayed senescence of transgenic Pssu-ipt tobacco. Control Nicotiana tabacum L. (cv. Petit Havana SR1) and transgenic tobacco with the ipt gene under the control of the promoter of small subunit of Rubisco (Pssu-ipt) were both grown either as grafts on control rootstocks or as rooted plants. Both control plant types showed a decline in total content of CKs with proceeding plant senescence. Contrary to this both transgenic plant types exhibited at least ten times higher content of CKs than controls and a significant increase of CK contents throughout the ontogeny with maximal values in the later stages of plant development. Significantly higher portion of O-glucosides was found in both transgenic plant types compared to control ones. In transgenic plants, zeatin and zeatin riboside were predominant type of CKs. Generally, Pssu-ipt tobacco exhibited elevated activities of antioxidant enzymes compared to control tobacco particularly in the later stages of plant development. While in control tobacco activity of glutathione reductase (GR) and superoxide dismutase (SOD) showed increasing activity up to the onset of flowering and then gradually decreased, in both transgenic types GR increased and SOD activity showed only small change throughout the plant ontogeny. Ascorbate peroxidase (APOD) was stimulated in both transgenic types. The manifold enhancement of syringaldazine and guaiacol peroxidase activities was observed in transgenic grafts throughout plant ontogeny in contrast to control and transgenic rooted plants, where the increase was found only in the late stages. Electron microscopic examination showed higher number of crystallic cores in peroxisomes and abnormal interactions among organelles in transgenic tobacco in comparison with control plant. The overproduction of cytokinins resulted in the stimulation of activities of AOE throughout the plant ontogeny of transgenic Pssu-ipt tobacco.