紫外线对大鼠晶状体抗氧化酶mRNA表达水平的影响

为探讨紫外线对晶状体的损伤机制,用ＲＴ－ＰＣＲ方法（ｒｅｖｅｒｓｅｔｒａｎｓｃｒｉｐｔｉｏｎ－ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ,反转录聚合酶链反应）,研究经紫外线照射后大鼠晶状体抗氧化相关酶,包括铜锌－超氧化物歧化酶（ｃｏｐｐｅｒ－ｚｉｎｃ－ｓｕｐｅｒｏｘｉｄｅｄｉｓｍｕｔａｓｅ,Ｃｕ－Ｚｎ－ＳＯＤ）,谷胱甘肽过氧化物酶（ｇｌｕ－ｔａｔｈｉｏｎｅｐｅｒｏｘｉｄａｓｅ,ＧＳＨ－Ｐｘ）和过氧化氢酶（ｃａｔａｌａｓｅ,ＣＡＴ）等ｍＲＮＡ的表达．结果显示,短时间的照射（２～５ｍｉｎ）,抗氧化相关酶的ｍＲＮＡ表达水平有增高表现,随后其ｍＲＮＡ表达水平开始下降,１５ｍｉｎ时抗氧化相关酶ｍＲＮＡ的表达下降更为明显,与对照组相比有非常显著性差异（Ｐ＜０．００１）．照射后２４ｈ,抗氧化相关酶的ｍＲＮＡ表达有不同程度的恢复;照射后４８ｈ,其ｍＲＮＡ表达水平基本恢复,与对照组相比没有显著性差异．从而从基因水平上初步探讨了紫外线的氧化损伤机制     

Comparison of antioxidant enzymes in saliva of elderly smokers and non-smokers

Objectives: This study was designed to compare the levels of copper/zinc superoxide dismutase (Cu/Zn SOD), peroxidase (POx) and glutathione peroxidase (GSH‐Px) in saliva of smokers and those in saliva of non‐smokers. Methods: Unstimulated saliva samples were collected from 88 elderly males (65 years old or over) who visited a private dental clinic. Forty‐four subjects were current smokers (more than 20 cigarettes daily for at least 30 years) and 44 were non‐smokers. The levels of salivary thiocyanate, Cu/Zn SOD, GSH‐Px, and POx activity were measured using standard procedures. Results: The mean levels of salivary thiocyanate (SCN^?) and SOD were significantly higher (p < 0.01) in the smoking group than in the non‐smoking group, whereas the specific activity levels of POx and GSH‐Px were significantly higher (p < 0.05) in the non‐smoking group than in the smoking group. Significant correlation coefficients were found between the levels of SCN^? and SOD (r = 0.37, p < 0.001). In the non‐smoking group, a significant positive association was found between specific activity of POx and age (r = 0.33, p < 0.05). Conclusion: Measurement of SCN^? and Cu/Zn SOD in human saliva might be useful for estimating the level of oxidative stress caused by cigarette smoke. Despite increased H₂O₂ level as a defense system induced by SOD, detoxification of H₂O₂ might be deteriorated in the oral cavity of elderly smokers.     

Manganese-excess induces oxidative stress,lowers the pool of antioxidants and elevates activities of key antioxidative enzymes in rice seedlings

Manganese (Mn) is an essential element for plant growth but in excess, specially in acidic soils, it can become phytotoxic. In order to investigate whether oxidative stress is associated with the expression of Mn toxicity during early seedling establishment of rice plants, we examined the changes in the level of reactive oxygen species (ROS), oxidative stress induced an alteration in the level of non-enzymic antioxidants and activities of antioxidative enzymes in rice seedlings grown in sand cultures containing 3 and 6 mM MnCl₂. Mn treatment inhibited growth of rice seedlings, the metal increasingly accumulated in roots and shoots and caused damage to membranes. Mn treated plants showed increased generation of superoxide anion (O₂ ^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) and decline in protein thiol. The level of nonprotein thiol, however, increased due to Mn treatment. A decline in contents of reduced ascorbate (AsA) and glutathione (GSH) as well as decline in ratios of their reduced to oxidize forms was observed in Mn-treated seedlings. The activities of antioxidative enzymes superoxide dismutase (SOD) and its isoforms Mn SOD, Cu/Zn SOD, Fe SOD as well as guaiacol peroxidase (GPX) increased in the seedlings due to Mn treatment however, catalase (CAT) activity increased in 10 days old seedlings but it declined by 20 days under Mn treatment. The enzymes of Halliwell-Asada cycle, ascorbate peroxidase (APX) monodehydoascorbate reductase (MDHAR), dehyroascorbate reductase (DHAR) and glutathione reductase (GR) increased significantly in Mn treated seedlings over controls. Results suggest that in rice seedlings excess Mn induces oxidative stress, imbalances the levels of antioxidants and the antioxidative enzymes SOD, GPX, APX and GR appear to play an important role in scavenging ROS and withstanding oxidative stress induced by Mn.     

The antioxidant defense system of isolated guinea pig Leydig cells

Utilization of highly enriched preparations of steroidogenic Leydig cells have proven invaluable for studying the direct effects of various hormones and agents on Leydig cell functionin vitro. However, recent work indicates that isolated Leydig cells are often subjected to oxygen (O₂) toxicity when cultured at ambient (19%) oxygen concentrations. Because intracellular antioxidants play an important role in protecting cells against oxygen toxicity, we have investigated the intracellular antioxidant defense system of isolated Leydig cells. The cellular levels of several antioxidants including catalase, glucose-6-phosphate dehydrogenase (G-6-PDH), superoxide dismutase (SOD) of the Cu/Zn & Mn variety, glutathione peroxidase, glutathione reductase and total glutathione were quantitated using enriched populations of Leydig cells isolated from adult male guinea pig testes. Compared to whole testicular homogenates, Leydig cells contained significantly (P<0.01) less G-6-PDH, total SOD, glutathione reductase and total glutathione, but significantly (P<0.001) more glutathione peroxidase. Compared to hepatic values previously reported in the guinea pig, Leydig cells contain nearly 400 times less catalase, about 14 times less glutathione peroxidase and almost 11 times less glutathione reductase. Since G-6-PDH and glutathione reductase are both necessary to regenerate reduced gluthathione (GSH) which couples with glutathione peroxidase to breakdown hydrogen peroxide (H₂O₂) under normal conditions, it is plausible that the oxygen toxicity observed in isolated Leydig cells is due to the intracellular accumulation of H₂O₂. Using the dichlorofluorescin diacetate (DCF-DA) assay, we found that Leydig cells incubated in the presence of 19% O₂ produced significantly (P<0.001) higher levels of H₂O₂ with time in culture compared to Leydig cells maintained at 3% O₂. These results support the hypothesis that the increased susceptibility of isolated Leydig cells to oxygen toxicity may be due, in part, to decreased amounts of certain antioxidant defenses and an increased production of the reactive oxygen species H₂O₂.     

Antioxidants activities and concentration of selenium,zinc and copper in preterm and IUGR human placentas

The aim of this study was to examine changes in activities of cytochrome c oxidase (CCO), glucose-6-phosphate dehydrogenase (G6PDH), Cu–Zn superoxide dismutase (Cu–Zn SOD), glutathione peroxidase (GSH-Px), glutathione (GSH) levels and copper (Cu), zinc (Zn) and selenium (Se) concentrations, and to assess the possible differences between preterm placentas, placentas from term pregnancies complicated by intrauterine growth restriction (IUGR) and full-term control placentas.The enzyme activities and the level of GSH decreased in IUGR and preterm placentas in comparison with the control group. CCO activity and GSH level in preterm placentas were markedly lower compared with the IUGR (P<0.01; P<0.05) and control (P<0.01; P<0.05) placentas, respectively. In IUGR placentas the level of Cu was reduced by 23% (P<0.05) and Zn by 37%. In preterm placentas the level of Cu was reduced by 19% and Zn by 42%. Se level in IUGR and preterm placentas was higher (P<0.05) by 28% and 32% than in control group, respectively. The strong relation was observed between birth weight and CCO activity, birth weight and Cu–Zn SOD activity, and a low level of Zn and Cu influenced the birth weight especially in IUGR cases. Moreover, the strong inverse correlation between Se level and birth weight, Se level and placental weight and Se level and CCO activity are new findings.     

不同海拔青海云杉与祁连圆柏叶片抗氧化系统

以祁连山寺大隆林区连续海拔梯度(2 665~3 365 m)上青海云杉(Picea crassifolia)和祁连圆柏(Sabina przewalskii)为材料, 测定叶片中抗氧化保护系统的变化, 探讨常绿木本植物抗氧化系统对高山极端环境的适应机制。结果显示, 祁连圆柏和青海云杉叶片中丙二醛(MDA)含量均与海拔高度呈正相关, 相同海拔上青海云杉MDA含量极显著高于祁连圆柏(p＜0.01)。随海拔升高, 两树种抗氧化保护酶超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)活性和非酶促抗氧化剂脯氨酸(Pro)、抗坏血酸(AsA)、还原性谷胱甘肽(GSH)含量均呈明显增加趋势。青海云杉叶片的AsA水平高于祁连圆柏, 但对海拔变化的敏感性较低; 祁连圆柏的GSH、Pro水平及其对海拔变化的敏感性均高于青海云杉。结果表明, 研究区青海云杉所受过氧化伤害较祁连圆柏更严重, 但两树种清除O₂^-·的能力相当而主要负责分解H₂O₂的酶种有所不同: 祁连圆柏中为POD, 青海云杉中则为CAT、APX和GR, AsA-GSH循环系统在青海云杉活性氧清除中的作用强于在祁连圆柏中, 祁连圆柏的活性氧清除物质可能以Pro为主。     

Activities of Hydrogen Peroxide-Scavenging Enzymes in Germinating Wheat Seeds

During imbibition and germination of wheat (Triticum aestivum)in the dark over 72 h, activities of the enzymes of the ascorbate(AsA)-dependent H₂O₂-scavenging pathway, AsA peroxidase, monodehydroascorbate(MDAsA) reductase, dehydroascorbate (DHAsA) reductase and glutathione(GSSG) reductase as well as superoxide dismutase (SOD), catalaseand guaiacol peroxidase were determined both in whole grainsand in isolated embryos and endosperm. With the exception of DHAsA reductase, activities of the otherenzymes assayed increased in germinating seeds, especially duringradicle emergence (between 24–48 h of imbibition). Theseincreases, particularly for AsA peroxidase, were much higherin the embryo than in the endosperm. Within 72 h of imbibition,activities per seed increased 116-fold for AsA peroxidase, 19-foldfor guaiacol peroxidase, 5-fold for catalase and only 1·4-foldfor SOD. In contrast to the decreases in DHAsA reductase, theother AsA recycling enzyme, MDAsA reductase, increased 5-foldwithin 72 h. The results indicate that, in wheat seeds, imbibition and germinationis associated with enhanced cellular capacity to detoxify H₂O₂.For this detoxification the operation of AsA peroxidase togetherwith the AsA-regenerating enzymes appears to be of particularimportance. Key words: Ascorbate peroxidase, germination, hydrogen peroxide detoxification, inhibition, wheat     

The balance between Cu,Zn-superoxide dismutase and catalase affects the sensitivity of mouse epidermal cells to oxidative stress.

Oxidants are toxic, but at low doses they can stimulate rather than inhibit the growth of mammalian cells and play a role in the etiology of cancer and fibrosis. The effect of oxidants on cells is modulated by multiple interacting antioxidant defense systems. We have studied the individual roles and the interaction of Cu,Zn-superoxide dismutase (SOD) and catalase (CAT) in transfectants with human cDNAs of mouse epidermal cells JB6 clone 41. Since only moderate increases in these enzymes are physiologically meaningful, we chose the following five clones for in-depth characterization: CAT 4 and CAT 12 with 2.6-fold and 4.2-fold increased catalase activities, respectively, SOD 15 and SOD 3 with 2.3-fold and 3.6-fold increased Cu,Zn-SOD activities, respectively, and SOCAT 3 with a 3-fold higher catalase activity and 1.7-fold higher Cu,Zn-SOD activity than the parent JB6 clone 41. While the increases in enzyme activities were moderate, the human cDNAs were highly expressed in the transfectants. As demonstrated for the clone SOD 15, this discordance between message concentrations and enzyme activities may be due to the low stability of the human Cu,Zn-SOD mRNA in the mouse recipient cells. According to immunoblots the content of Mn-SOD was unaltered in the transfectants. While the activities of glutathione peroxidase were comparable in all strains, the concentrations of reduced glutathione (GSH) were significantly lower in SOD 3 and SOD 15. This decrease in GSH may reflect a chronic prooxidant state in these Cu,Zn-SOD overproducers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Salinity, oxidative stress and antioxidant responses in shoot cultures of rice

When shoot cultures derived from salt-sensitive Oryza sativavar. Taipei 309 were grown at 25C in medium containing 0.35M NaCl, responses to possible oxidative stress in the earlystages of exposure were observed. Overall levels of Mn-superoxidedismutase activity, Cu, Zn-superoxide dismutase activity andH₂O₂ were significantly elevated. After 1 d there was a notabledecline in tissue concentrations of GSH and a correspondingincrease in GSSG. However, after a further day, concentrationsof GSH and GSSG returned to concentrations normally encounteredin control cultures. Activities of ascorbate peroxidase andcatalase were similar whether the shoots were grown in the presenceor absence of NaCl. In contrast, there was an early increasein glutathione reductase activity in NaCl-exposed cultures,and no indication of extensive increases in lipid peroxidation.Thus although some indications of oxidative stress accompanyexposure of this salt-sensitive rice variety to salinity, mechanismsappear to exist within its shoot tissue to permit the toleranceof such oxidative stress. Key words: Salinity stress, hydrogen peroxide, glutathione, antioxidant enzymes, Oryza sativa     

Effects of fluoride on hepatic antioxidant system and transcription of Cu/Zn SOD gene in young pigs

Thirty-two barrows (Duroc x Landrace x Yorkshire) were randomly divided into four groups, each of which included eight pigs. The groups received the same basal diet supplemented with 0, 100, 250 and 400mg/kg fluoride, respectively. The malondialdehyde (MDA) and glutathione (GSH) levels, antioxidant enzymes activities and zinc/copper superoxide dismutase (Cu/Zn SOD) mRNA content in the liver were determined to evaluate the fluoride hepatic intoxication. Results showed the increased lipid peroxides (LPO) level and the reduced GSH content, along with a concomitant decrease in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px). Moreover, the level of hepatic Cu/Zn SOD mRNA was also significantly reduced. We suggest the mechanism of fluoride injuring the liver as follows: fluoride causes a decrease in Cu/Zn SOD mRNA and the reduced activities of antioxidant enzymes, leads to the declined ability of scavenging free radicals with excessive production of LPO, which seriously damages the hepatic structure and function.     

Antioxidant status of the rat nasal cavity

Despite extensive interest in the rodent nasal cavity as a target organ for toxicity, there is very limited information regarding nasal defenses against oxidative stress and xenobiotic-derived oxidants. Using immunohistochemistry, we have examined the distribution of Cu,Zn and Mn superoxide dismutase (SOD), catalase, glutathione (GSH) peroxidase, and DT-diaphorase in rat nasal tissues. In addition, we have determined the concentrations of ascorbate and alpha-tocopherol and the activities of SOD (combined Cu,Zn and Mn forms), catalase, GSH peroxidase, GSH reductase, and DT-diaphorase in nasal respiratory epithelium (RE), olfactory epithelium (OE), and in lung. Immunohistochemistry demonstrated that all four enzymes were similarly distributed, with the greatest staining intensity in dorsal-medial regions of the nasal cavity. In respiratory epithelium, ciliated columnar cells and subepithelial glands stained positively, while in olfactory tissue the enzymes were detected in the sustentacular cells and Bowman's glands. With the exception of SOD, enzyme activities were higher in RE than OE, while concentrations of ascorbate and alpha-tocopherol were higher in OE than RE. With the exception of catalase, nasal activities were either higher than or comparable to those of the lung. Thus, the rat nasal cavity appears to be well protected against oxidative damage.     

Exogenous sodium nitroprusside and glutathione alleviate copper toxicity by reducing copper uptake and oxidative damage in rice (Oryza sativa L.) seedlings

Nitric oxide (NO) and glutathione (GSH) regulate a variety of physiological processes and stress responses; however, their involvement in mitigating Cu toxicity in plants has not been extensively studied. This study investigated the interactive effect of exogenous sodium nitroprusside (SNP) and GSH on Cu homeostasis and Cu-induced oxidative damage in rice seedlings. Hydroponically grown 12-day-old seedlings were subjected to 100 μM CuSO₄ alone and in combination with 200 μM SNP (an NO donor) and 200 μM GSH. Cu exposure for 48 h resulted in toxicity symptoms such as stunted growth, chlorosis, and rolling in leaves. Cu toxicity was also manifested by a sharp increase in lipoxygenase (LOX) activity, lipid peroxidation (MDA), hydrogen peroxide (H₂O₂), proline (Pro) content, and rapid reductions in biomass, chlorophyll (Chl), and relative water content (RWC). Cu-caused oxidative stress was evident by overaccumulation of reactive oxygen species (ROS; superoxide (O₂ ^?–) and H₂O₂). Ascorbate (AsA) content decreased while GSH and phytochelatin (PC) content increased significantly in Cu-stressed seedlings. Exogenous SNP, GSH, or SNP?+?GSH decreased toxicity symptoms and diminished a Cu-induced increase in LOX activity, O₂ ^?–, H₂O₂, MDA, and Pro content. They also counteracted a Cu-induced increase in superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and glyoxalase I and glyoxalase II activities, which paralleled changes in ROS and MDA levels. These seedlings also showed a significant increase in catalase (CAT), glutathione peroxidase (GPX), dehydroascorbate reductase (DHAR), glutathione S-transferase (GST) activities, and AsA and PC content compared with the seedlings stressed with Cu alone. Cu analysis revealed that SNP and GSH restricted the accumulation of Cu in the roots and leaves of Cu-stressed seedlings. Our results suggest that Cu exposure provoked an oxidative burden while reduced Cu uptake and modulating the antioxidant defense and glyoxalase systems by adding SNP and GSH play an important role in alleviating Cu toxicity. Furthermore, the protective action of GSH and SNP?+?GSH was more efficient than SNP alone.     

Glutathione metabolism in Urtica dioica in response to cadmium based oxidative stress

To investigate the antioxidative response of glutathione metabolism in Urtica dioica L. to a cadmium induced oxidative stress, activities of glutathione reductase (GR), glutathione-S-transferase (GST), and glutathione peroxidase (GSH-Px), content of reduced (GSH) and oxidized (GSSG) glutathione, lipid peroxidation (LPO), and also accumulation of Fe, Zn, Mn, Cu besides Cd were determined in the roots, stems, and leaves of plants exposed to 0 (control), 0.045, and 0.09 mM CdCl₂ for 58 h. Whereas the Cd content continuously increased in all organs, the Fe, Zn, Mn, and Cu content decreased in dependence on the applied Cd concentration and incubation time. The Cd treatment resulted in increased GR and GST activities in all organs, however, GSH-Px activity was dependent on Cd concentration and plant organ. The GSH/GSSG ratio maintained above the control level in the stems at both Cd concentrations. The LPO was generally close to the control values in the roots and stems but it increased in the leaves especially at 0.09 mM Cd.     

Comparative study of alleviating effects of GSH, Se and Zn under combined contamination of cadmium and chromium in rice (Oryza sativa)

A hydroponic experiment was conducted to study the ameliorative effects of separate or combined application of exogenous glutathione (GSH), selenium (Se) and zinc (Zn) upon 20 μM cadmium (Cd) plus 20 μM chromium (Cr) heavy metal stress (HM) in rice seedlings. The results showed that HM caused a marked reduction in seedling height, chlorophyll content (SPAD) and biomass, and activities of catalase (CAT) and ascorbate peroxidase (APX) in leaves and H⁺-ATPase in roots/leaves, but elevated superoxide dismutase (SOD) and guaiacol peroxidase (POD) activities in leaves with elevated malondialdehyde (MDA) accumulation both in leaves and roots over the control. The best mitigation effect was recorded in HM+GSH+Zn and HM+GSH (addition of GSH+Zn and GSH to HM solution), which greatly alleviated HM-induced growth inhibition and oxidative stress. Compared with HM alone, HM+GSH and HM+GSH+Zn markedly reduced Cr uptake and translocation but not affected Cd concentration; improved H⁺-ATPase activity and Fe, Zn, Mn uptake and translocation, and repressed MDA accumulation. Meanwhile exogenous GSH and GSH+Zn counteracted HM-induced response of antioxidant enzymes, via suppressing HM-induced dramatic increase of root/leaf SOD and leaf POD activities, and elevating stress-depressed leaf APX and leaf/root CAT activities.     

Over-expression of chloroplast-targeted Mn superoxide dismutase in cotton (Gossypium hirsutum L., cv. Coker 312) does not alter the reduction of photosynthesis after short exposures to low temperature and high light intensity

Transgenic cotton plants from several independently-transformed lines expressing a chimeric gene encoding a chloroplast-targeted Mn superoxide dismutase (SOD) from tobacco exhibit a three-fold increase in the total leaf SOD activity, strong Mn SOD activity associated with isolated chloroplasts, and a 30% and 20% increase in ascorbate peroxidase and glutathione reductase activities, respectively. The Mn SOD plants did exhibit a slightly enhanced protection against light-mediated, paraquat-induced cellular damage but only at 0.3 µM paraquat. In addition, photosynthetic rates at 10°C and 15°C were similar to those of controls, and the immediate recovery of photosynthesis after a 35-min exposure to 5°C and full sun was only slightly better than that for wild-type plants. The recovery for longer exposure times was comparable for both genotypes as was the deactivation of the H₂O₂-sensitive, Calvin-cycle enzyme, stromal fructose 1,6-bisphosphatase (FBPase). Compared to the controls, Mn SOD plant leaves in full sun prior to chilling stress had a lower activation of FBPase, a higher ratio of oxidized to reduced forms of ascorbate, and a higher total glutathione content. After 35 min at 5°C in full sunlight, total glutathione had risen in control leaves to 88% of the Mn SOD plant values, and oxidized to reduced ascorbate ratios were higher for both genotypes. However, an 80% increase in the ratio of oxidized to reduced glutathione occurred for Mn SOD plant leaves with no change for controls. This increased demand on the ascorbate-glutathione cycle is circumstantial evidence that high Mn SOD activity in the chloroplast leads to increased H₂O₂ pools that could, in some manner, affect photosynthetic recovery after a stress period. We postulate that the pool sizes of reduced ascorbate and glutathione may restrict the ability of the ascorbate-glutathione cycle to compensate for the increased activity of SOD in cotton over-producing mitochondrial Mn SOD in chloroplasts during short-term chilling/high light stress.     

Copper-induced changes in the growth, oxidative metabolism, and saponin production in suspension culture roots of Panax ginseng in bioreactors

Roots of Panax ginseng exposed to various concentrations of Cu (0.0, 5, 10.0, 25.0, and 50.0 μM) accumulated high amounts of Cu in a concentration-dependent and duration-dependent manner. Roots treated with 50 μM Cu resulted in 52% and 89% growth inhibition after 20 and 40 days, respectively. Saponin synthesis was stimulated at a Cu concentration between 5 and 25 μM but decreased at 50 μM Cu. Malondialdehyde content (MDA), lipoxygenase activity (LOX), superoxide ion (O₂ ^•−) accumulation, and H₂O₂ content at 5 and 10 μM Cu-treated roots were not increased but strongly increased at 50 μM Cu resulting in the oxidation of ascorbate (ASC) and glutathione (GSH) to dehydroascorbate (DHA) and glutathione disulfide (GSSG), respectively indicating a clear oxidative stress. Seven well-resolved bands of superoxide dismutase (SOD) were detected in the gel and an increase in SOD activity seemed to be mainly due to the induction of Fe-SOD 3. Five to 10 μM Cu slightly induced activity of ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR), guaiacol peroxidase (G-POD) but inhibited monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR) enzyme activities. No changes in catalase (CAT) activity and in activity gel were found up to 25 μM Cu, but both G-POD and CAT activities were inhibited at 50 μM Cu. Glutathione metabolism enzymes such as γ-glutamylcysteine synthetase (γ-GCS), glutathione-S-transferase (GST), and glutathione peroxidase activities (GPx) were activated at 5 and 10 μM Cu but were strongly inhibited at 50 μM Cu due to the Cu accumulation in root tissues. The strong depletion of GSH at 50 μM Cu was associated to the strong induction of γ-glutamyltranspeptidase (γ-GGT) activity. These results indicate that plant could grow under Cu stress (5–25 μM) by modulating the antioxidant defense mechanism for combating Cu induced oxidative stress.     

Overexpression of Superoxide Dismutase Protects Plants from Oxidative Stress (Induction of Ascorbate Peroxidase in Superoxide Dismutase-Overexpressing Plants)

Photosynthesis of leaf discs from transgenic tobacco plants (Nicotiana tabacum) that express a chimeric gene that encodes chloroplast-localized Cu/Zn superoxide dismutase (SOD+) was protected from oxidative stress caused by exposure to high light intensity and low temperature. Under the same conditions, leaf discs of plants that did not express the pea SOD isoform (SOD-) had substantially lower photosynthetic rates. Young plants of both genotypes were more sensitive to oxidative stress than mature plants, but SOD+ plants retained higher photosynthetic rates than SOD- plants at all developmental stages tested. Not surprisingly, SOD+ plants had approximately 3-fold higher SOD specific activity than SOD- plants. However, SOD+ plants also exhibited a 3- to 4-fold increase in ascorbate peroxidase (APX) specific activity and had a corresponding increase in levels of APX mRNA. Dehydroascorbate reductase and glutathione reductase specific activities were the same in both SOD+ and SOD- plants. These results indicate that transgenic tobacco plants that overexpress pea Cu/Zn SOD II can compensate for the increased levels of SOD with increased expression of the H2O2-scavenging enzyme APX. Therefore, the enhancement of the active oxygen-scavenging system that leads to increased oxidative stress protection in SOD+ plants could result not only from increased SOD levels but from the combined increases in SOD and APX activity.     

Antioxidant responses of pea genotypes to zinc deficiency

The effects of Zn deficiency on antioxidant responses of two pea (Pisum sativum L.) genotypes, a Zn-efficient IPFD-99-13 and Zn-inefficient KPMR-500, grown in sand culture were studied. In the pea genotype KPMR-500, Zn deficiency decreased dry matter yield, tissue Zn concentration, and antioxidant enzyme activities istronger than in the genotype IPFD-99-13. Genotype IPFD-99-13 developed more efficient antioxidant system to scavenge ROS than genotype KPMR-500. Zinc deficiency produced oxidative damage to pea genotypes due to enhanced accumulation of TBARS and H₂O₂ and decreased activities of antioxidant enzymes (Cu/Zn superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX)). In the leaves of IPFD-99-13 genotype, the higher activity of ROS-scavenging enzyme, e.g., SOD, CAT, POD, and glutathione reductase, and antioxidants, such as ascorbate and non-protein thiols, led to the lower accumulation of H₂O₂ and lipid peroxides. These results suggest that, by maintaining an efficient antioxidant defense system, the IPFD-99-13 genotype shows a lower sensivity to Zn deficiency than the KPMR-500 genotype.     

Comparison of intracellular glutathione and related antioxidant enzymes: Impact of two glycosylated whey hydrolysates

The effects of two glycosylated whey hydrolysates (GWH-Gal A and GWH-Gal B) on glutathione (GSH) and related antioxidant enzymes in SGC-7901 cells were evaluated. Two whey glycosylated hydrolysates promoted an increase in reduced glutathione (GSH) in normal SGC-7901 cells. GSH, glutathione peroxidase (GPx), γ-glutamine cysteine synthetaase (γ-GCS), and catalase (CAT) at 1.0 and 2.0 mg/mL in normal SGC-7901 cells were higher in the GWH-Gal A group than in the GWH-Gal B group (P < 0.05). Compared with GWH-Gal B, GWH-Gal A more strongly inhibited decreases in intracellular GSH, GPx, γ-GCS, CAT, and superoxide dismutase (SOD) in H₂O₂-induced SGC-7901 cells. Compared with GWH-Gal B, GWH-Gal A at 1.0 and 2.0 mg/mL effectively inhibited increases in lactate dehydrogenase (LDH) and malondialdehyde (MDA) in H₂O₂-induced SGC-7901 cells (P < 0.05). Therefore, GSH content and related antioxidant enzyme activity levels (GPx, γ-GCS, CAT, SOD) in both normal and H₂O₂-induced SGC-7901 cells were considerably stronger in the GWH-Gal A group than in the GWH-Gal B group.     

Antioxidant defences of the apoplast

Summary The apoplast of barley and oat leaves contained superoxide dismutase (SOD), catalase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase, and glutathione reductase activities. The activities of these enzymes in the apoplastic extracts were greatly modified 24 h after inoculation with the biotrophic fungal pathogenBlumeria graminis. The quantum efficiency of photosystem II, which is related to photosynthetic electron transport flux, was comparable in inoculated and healthy leaves during this period. Apoplastic soluble acid invertase activity was also modified in inoculated leaves. Inoculation-dependent increases in apoplastic SOD activity were observed in all lines. Major bands of SOD activity, observed in apoplastic protein extracts by activity staining of gels following isoelectric focusing, were similar to those observed in whole leaves but two additional minor bands were found in the apoplastic fraction. The apoplastic extracts contained substantial amounts of dehydroascorbate (DHA) but little or no glutathione (GSH). Biotic stress decreased apoplastic ascorbate and DHA but increased apoplastic GSH in resistant lines. The antioxidant cycle enzymes may function to remove apoplastic H₂O₂ with ascorbate and GSH derived from the cytoplasm. DHA and oxidized glutathione may be reduced in the apoplast or returned to the cytosol for rereduction.Abbreviations AA reduced ascorbate - APX ascorbate peroxidase - DHA dehydroascorbate (oxidised ascorbate) - DHAR dehydroascorbate reductase - G6PDH glucose-6-phosphate dehydrogenase - GSH reduced glutathione - GSSG glutathione disulphide - GR glutathione reductase - MDHA monodehydroascorbate - MDHAR monodehydroascorbate reductase - SOD superoxide dismutase