原虫硫氧还蛋白研究进展

硫氧还蛋白(Trx)是一种广泛存在于原核生物和真核生物中的小分子蛋白质,它和硫氧还蛋白还原酶(TrxR)、还原型烟酰胺腺嘌呤二核苷酸磷酸Ⅱ(NADPH)共同构成了硫氧还蛋白系统,是硫氧还蛋白系统的重要组成部分。顶复门原虫作为胞内寄生虫,自身缺乏抗氧化的过氧化氢酶和谷胱甘肽过氧化物酶,依靠硫氧还蛋白系统抵抗宿主的氧化作用。论文介绍了原虫Trx的结构,从抗氧化应激和蛋白结合等方面阐述了Trx的生物学活性及功能,介绍了Trx及其抑制剂在抗肿瘤等方面的研究进展,为抗原虫药物的发现提供参考。     

Effect of acute salinomycin-tiamulin toxicity on the lipid peroxide and antioxidant status of broiler chicken.

The combined effect, if any, of salinomycin poisoning and salinomycin-tiamulin interaction on lipid-peroxidative processes and the antioxidative defence system of the liver was studied in domestic fowl. Male broilers (28-day-old), reared on a diet containing 60 mg/kg salinomycin, were treated intraoesophageally with salinomycin (140 mg/kg body mass) or tiamulin (50 mg/kg body mass). Malondialdehyde, reduced glutathione and cytochrome P-450 concentrations as well as glutathione peroxidase and catalase activities of the liver were determined. Liver malondialdehyde concentration rose in the salinomycin-treated group while the amount of cytochrome P-450 increased in both groups treated. Glutathione concentration and glutathione peroxidase activity of the liver decreased rapidly but hepatic catalase activity increased in both groups after the treatment. Manifestation of the effect exerted by salinomycin and salinomycin-tiamulin on lipid-peroxidative processes nearly coincided with the onset of clinical signs and preceded the increase of hepatic cytochrome P-450 concentration. According to the results, the background of the previously reported incompatibility between salinomycin and tiamulin is the synergistic effect exerted on the antioxidant (glutathione) system.     

Association Between ETFA Genotype and Activity of Superoxide Dismutase,Catalase and Glutathione Peroxidase in Cryopreserved Sperm of Holstein–Friesian Bulls

The aim of this study was to determine whether C/T missense mutation within the ETFA gene is associated with sperm antioxidant enzymatic activity. One hundred and twenty Holstein–Friesian bulls were genotyped by the PCR‐RFLP technique (MwoI). Commercial straws of frozen‐thawed semen were used to evaluate the activity of three antioxidant enzymes: superoxide dismutase, catalase and glutathione peroxidase. Among all bulls investigated, genotype CT was the most frequent (44.2%), in comparison with CC (42.5%) and TT (13.3%). Significant differences in glutathione peroxidase activity were observed between homozygous individuals (CC vs TT) with heterozygous CT having intermediate values. Dismutase activity was significantly associated with ETFA genotype, although only bulls with the CT genotype were significantly different from bulls carrying the CC genotype. The activity of catalase showed a similar trend (but was not statistically significant). In conclusion, we found that bulls with the ETFA TT genotype produce sperm with the highest glutathione peroxidase activity and can therefore be more efficiently protected from reactive oxygen. The mechanism of this interaction needs to be elucidated in future research.     

Enzymatic antioxidant defense in isolated rat hepatocytes exposed to cadmium

The aim of the study was the evaluation of cadmium effects on the activity of antioxidant enzymes in rat hepatocytes. The studies were conducted with isolated rat hepatocytes incubated for 1 or 2 hours in a modified (deprived of carbonates with phosphates) Williams' E medium (MWE) in the presence of cadmium chloride (25, 50 and 200 microM). Hepatocytes incubated in the MWE medium without cadmium chloride were used as a control. The application of the modified Williams' E medium allowed for the appearance of cadmium compounds in a soluble form that is indispensable for suitable estimation of its toxic action. There were evaluated markers of the oxidative stress such as: concentration of thiobarbiturate reactive substances (TBARS)--proportional to the level of lipid peroxidation, concentration of reduced glutathione (GSH), and the activity of antioxidant enzymes, including superoxide dismutase (SOD1 and SOD2), catalase (CAT), total glutathione peroxidase (GSHPx), selenium--dependent glutathione peroxidase (SeGSHPx), glutathione transferase (GST) and glutathione reductase (GSHR). Alterations of antioxidant enzymes activity, the level of TBARS and GSH in isolated rat hepatocytes caused by cadmium in vitro, were shown to depend on the concentration and time of exposure of cells to this metal. The increased level of TBARS and GSH was observed as well as changes in the activity of antioxidant enzymes. The activity of SOD isoenzymes and CAT was increased, whereas GSHPx and GST were decreased. These results indicate that cadmium induces oxidative stress followed by alterations in the cellular antioxidant enzyme system in isolated rat hepatocytes.     

Effect of dietary supplementation with vitamins E and C on production performance, immune responses and antioxidant status of White Leghorn layers under tropical summer conditions

1. The effects of vitamin E (DL-alpha-tocopheryl acetate) and vitamin C (ascorbic acid) supplementation on performance, some immune variables and antioxidant status of White Leghorn layers (aged 44-56 weeks) exposed to tropical summer conditions were investigated. 2. Both vitamins E and C significantly improved the egg production, food conversion efficiency, antibody responses to inoculated sheep red blood cells (SRBC) and Newcastle disease virus (NDV) vaccine, lymphocyte proliferation in response to mitogen and activities of antioxidant enzymes red blood cell catalase (RBCC) and glutathione peroxidase (GSH-PX) in layers. Vitamin C increased the inflammatory response to phytohaemagglutinin-P (PHA-P) inoculation. 3. Supplemental vitamin C at 200 mg/kg diet significantly increased eggshell weight, eggshell thickness, and shell breaking strength. However, no effect of supplemental vitamin E on eggshell quality was observed. 4. Vitamins E and C, as well as interactions between them, significantly influenced the activities of glutathione reductase and lipid peroxidase. A combination of vitamin E at 125 IU/kg with vitamin C at 200 mg/kg diet had an additive effect on reducing the activity of oxidative enzyme lipid peroxidase (LP) and increasing the activity of antioxidant enzyme glutathione reductase (GSHR). 5. It is concluded that vitamin E (125 IU/kg) and vitamin C (200 mg/kg) could independently alleviate the effects of heat stress on production performance and immunological variables of layers. However, combination of both the vitamins at the above-mentioned concentrations is beneficial in eliciting higher antioxidant status in laying hens exposed to tropical summer conditions.     

SO2对家蚕血淋巴中谷胱甘肽含量及相关酶活性的影响

通过分析在SO2胁迫下家蚕5龄幼虫血淋巴中谷胱甘肽(GSH)含量及相关酶活性的变化,探讨SO2对家蚕的毒理作用机制。当对家蚕5龄幼虫每天以40 mg/m3SO2熏气刺激6 h,蚕体血淋巴中的GSH含量显著降低,雄蚕GSH的平均含量为49.8μmol/L,雌蚕为41.0μmol/L,分别为对照的73.80%和55.76%,而谷胱甘肽硫-转移酶(GST)活性显著升高,雄蚕GST的平均活性为31.86 U/L,雌蚕为31.80 U/L,分别高于对照46.15%和27.30%。与此同时,γ-谷氨酰转肽酶(γ-GT)活性在蚕体中也有不同程度的升高,其中在雄蚕体内的变化极为显著;谷胱甘肽过氧化酶(GSH-Px)和谷胱甘肽合成酶系的活性在蚕体内均无显著变化。以上结果表明:SO2刺激可对家蚕血淋巴中的GSH含量及其相关酶的活性产生影响,且雌、雄蚕对SO2的敏感性不同;GSH含量的减少及其相关酶活性的上升,说明SO2对蚕体的毒害作用与其降低蚕体抗氧化物质水平、削弱抗氧化防御系统有关。     

The relationship between serum biotin and oxidant/antioxidant activities in bovine lameness

Serum biotin concentrations, erythrocyte superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), reduced glutathione (GSH) and plasma thiobarbituric acid reactive substances (TBARS) were measured in 36 dairy cows, 18 of them were healthy and served as control. In the 18 cows with lameness problems, there were 5 cows with interdigital necrobacillosis, 5 cows with subsolar abscessation, 2 cows with solar ulcers, 2 cows with white line disease, 2 cows with chronic laminitis and 2 cows with septic arthritis. The degree of lameness was estimated to be slight in 3 cows, moderate in 11 cows and severe in 4 cows. Plasma fibrinogen levels and TBARS concentrations were increased significantly (P≤0.05) in lame cows compared to control group. The antioxidant enzymes GSH-Px, and CAT concentrations were increased significantly (P≤0.05) in lame cows. The level of reduced glutathione and the activity of SOD were significantly decreased in affected cows compared to healthy ones. Serum biotin levels in healthy cows ranged from 2.25 to 3.5ng/ml while in lame cows, biotin levels ranged from 1.17 to 2.3ng/ml. Biotin levels correlated positively with blood GSH (r=0.870, P≤0.05), (r=0.735, P≤0.05) and with GSH-Px (r=0.539, P≤0.05), (r=0.637, P≤0.05) and with SOD (r=0.637, P≤0.05), (r=0.449, P≤0.05) and with catalase (r=0.533, P≤0.05), (r=0.585, P≤0.05) in both healthy and lameness affected subjects, respectively.     

Interrelationships of erythrocyte glutathione peroxidase system enzymes in the fasted Beagle

Eight adult female Beagles were fasted for 21 days to investigate responses of the erythrocyte glutathione peroxidase system enzymes. Blood samples were collected once a week. Erythrocyte reduced glutathione was maintained within 10% of base-line value. Hexose monophosphate pathway dehydrogenases showed a significant inverse relationship to glutathione reductase (GR) activity. Further, active GR, as a percentage of total GR, increased as hexose monophosphate pathway dehydrogenase activity fell. A similar relationship between these enzymes has been reported in human glucose-6-phosphate dehydrogenase deficiency. Also, glutathione peroxidase showed an inverse linear response to erythrocytic reduced glutathione. Though the responses of this system are complex, they appear interrelated.     

奶牛性控精液与常规精液抗氧化物酶活性比较分析

通过比较常规精液和性控精液在活力、顶体完整率及精浆中四种抗氧化酶类活力的差异,对性控精液的品质进行综合评定。对用流式细胞仪分离后的性控精液和常规精液细管解冻(37.5℃,30s),分别分析精子活力和精子顶体完整率,并测定精浆中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)、谷胱甘肽还原酶(GR)和过氧化氢酶(CAT)的活性。结果表明性控精液精子在活力显著低于常规精液,且精浆中四种抗氧化酶类的活力也都显著低于常规精液。     

Activity of superoxide dysmutase, catalase and glutathione peroxidase in rats exposed to chlorpyrifos and erofloxacin

The aim of the study was to investigate the influence of administration of chlorpyrifos and/or enrofloxacin on the activity of chosen antioxidative enzymes i.e.: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in erythrocytes of rats. Chlorpyrifos was administered by stomach tube during 28 days at a dose of 3 mg/kg bw (0.02 LD50), and enrofloxacin was administered by stomach tube at a dose of 5 mg/kg bw during 3 subsequent days. It was stated that administration of enrofloxacin at applied dose did not cause any major changes in the activity of investigated antioxidative enzymes. The four-week exposure of rats to chlorpyrifos caused noticeable decrease in SOD and CAT activity in erythrocytes of rats at the beginning of the experiment (up to 24th hour) in comparison with the control group. The activity of GPx during all periods of the experiment was increased. In the group of animals in which both chlorpyrifos and enrofloxacin were applied, the profile of changes in activity of examined enzymes was similar to that one, which was observed after administration of chlorpyrifos exclusively, what may indicate lack of co-action between compounds used in the experiment.     

Molecular forms of selected antioxidant enzymes in dog semen--electrophoretical identification

The aim of the study was the electrophoretical identification of molecular forms of selected antioxidant enzymes in dog semen. Ejaculates to be studied were chosen from five dogs, aged from two to eight years. Polyacrylamide gel electrophoresis was carried out under non-denaturing conditions and then gels were stained for the activity of the following enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Sperm homogenates and all fractions (pre-spermatic, spermatic and post-spermatic) of dog ejaculate demonstrated one protein band with SOD activity characterized by low electrophoretic mobility. Based on the confirmed sensitivity to H2O2, it can be assumed that the detected SOD is an enzyme containing ions of Zn2+ and Cu2+ (Cu,Zn SOD). In sperm homogenates one protein band with GPx activity was characterized by high electrophoretic mobility, whereas in the spermatic and post-spermatic fractions of dog ejaculate three protein bands with different (low, medium and high) electrophoretic mobility were identified. CAT molecular forms were not found in either sperm homogenates or in the analyzed fractions of ejaculate.     

动物中活性氧的产生及清除机制

各种氧化应激使动物细胞中积累大量的活性氧，从而导致畜禽生产性能的下降。动物体内有效清除活性氧的保护机制分为酶促和非酶促两类。酶促系统包括超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH—Px)和过氧化物酶(POD)。非酶促系统包括抗坏血酸(Vc)、谷胱甘肽、维生素E、类胡萝卜紊等。在动物日粮中添加抗氧化剂来维持抗氧化平衡，是集约化管理条件下维持畜禽良好生产性能和繁殖性能的一个重要方法。     

Antioxidant profile of cataractous English Cocker Spaniels

Cataracts have been attributed to oxidative injury in proteins and lipids. Primary defenses that directly protect the lens against oxidative damage include small molecule antioxidants (vitamin C, vitamin E, glutathione and carotenoids) and antioxidant enzymes (superoxide dismutase, catalase, and the glutathione enzyme systems – glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase). In humans, low plasmatic levels of vitamin C, vitamin E and carotenoids have been associated with a high risk of senile cataracts. Dogs are more prone to develop cataracts. A decrease in antioxidant defenses could be responsible for increased lens oxidation and cataract development. In this study we report the levels of erythrocytic enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase) and plasma vitamin C as well as malondialdehyde, in normal and cataractous English Cocker Spaniel dogs. Plasma vitamin C levels were consistently lower in cataractous dogs (20.17 μ m  ± 8.2 μ m ) when compared with normal dogs (24.1 μ m  ± 9.4 μ m ). These results indicate a possibly decreased synthesis in vitamin C, leading to lower aqueous humor levels of this vitamin. Considering that vitamin C levels in the aqueous humor may be responsible for lens antioxidant maintenance, and that these levels are obtained from plasma secretion through the ciliary epithelium, decreased plasma levels may indicate a decrease in the antioxidant capacity of the aqueous humor.     

Tissue distribution of monomeric glutathione peroxidase in broiler chicks.

It is known that there are two kinds of enzymes which show glutathione peroxidase activity, 'classical' glutathione peroxidase and glutathione S-transferase. Recently, a third enzyme was found, monomeric glutathione peroxidase, in broiler chick liver cytosolic fraction. To gain an insight into the possible physiological role of the monomeric glutathione peroxidase, the distribution of this enzyme in other broiler tissues was studied. The monomeric glutathione peroxidase was found in all tissues examined and in erythrocytes. The percentage of the total glutathione peroxidase activity accounted for by the monomeric enzyme ranged from 4 per cent in erythrocytes to 28 per cent in liver. Livers from three avian and two mammalian species contained the monomeric glutathione peroxidase. The contribution of the monomeric enzyme to total glutathione peroxidase activity with cumene hydroperoxide was high in poultry livers, while only trace monomeric glutathione peroxidase activity was found in mammalian livers. Chick monomeric glutathione peroxidase showed high activity toward phospholipid hydroperoxide. Thus, monomeric glutathione peroxidase might be an important enzyme in reducing membrane lipid hydroperoxides in birds.     

High sensitivit to streptozotocin in herbivorous voles,Microtus arvalis,compared to mice

Herbivorous voles, Microtus arvalis, have characteristics similar to herbivores in that their hepatic glycolytic enzyme activities are relatively low. The effects of a single low dose (100 mg/kg body weight) of streptozotocin (STZ) in voles were studied and the difference in sensitivity to or toxicity of STZ in voles and C57BL/6 mice was compared. In voles which received STZ, the cumulative incidence of glycosuria reached 53% by 4 weeks after administration. The diabetic voles showed marked increas in their blood glucose and plasma free fatty acid concentrations and a significant decrease in plasma immunoreactive insulin concentrations. Their hepatic hexokinase, glucokinase, glutathione peroxidase and glutamate dehydrogenase activities decreased significantly and lesions were widely observed in the liver, kidney and pancreas. The activities of glutathione peroxidase, a scavenger of H₂O₂, decreased significantly in their liver and pancreas. These changes were not observed in C57BL/6 mice which received STZ. The higher sensitivity to and toxicity of STZ in voles than in mice are considered to be caused by the characteristically low activities of glycolytic enzymes and glutathione peroxidase in the tissues of voles. Voles may be a good model for studying the mechanisms of cytotoxicity by STZ in herbivorous animals.Abbreviations GK glucokinase - GIDH glutamate dehydrogenase - GSH-px glutathione peroxidase - HE haematoxylin-eosin - HK nexokinase - IRI immunoreactive insulin - STZ streptozotocin     

Amelioration of altered oxidant/antioxidant balance of Indian water buffaloes with subclinical mastitis by vitamins A, D3, E, and H supplementation

The effect of vitamins A, D₃, E, and H supplementation on oxidative stress indices in Indian water buffaloes suffering from subclinical mastitis was investigated. Changes in the total oxidant capacity (TOC), total antioxidant capacity (TAC), nitric oxide (NO), and activities of glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) in milk were evaluated before and after the supplementation of vitamins A, D₃, E, and H. The buffaloes suffering from subclinical mastitis revealed remarkable alterations in the milk oxidants/antioxidants balance shifted towards oxidative status. The buffaloes with subclinical mastitis revealed significantly (P?≤?0.01) higher TOC, NO contents, and CAT activity, while TAC content and GSH-Px activity were significantly (P?≤?0.01) lower in comparison with the healthy controls. However, SOD activity did not show any significant change. Supplementation of vitamins A, D₃, E, and H to these animals revealed significant (P?≤?0.01) reduction in TOC, NO, and CAT, while a significant (P?≤?0.01) increase in TAC and GSH-Px activity was also evident. From the present study, it may be concluded that supplementation of these vitamins can help ameliorate the altered milk oxidants/antioxidants balance towards normalcy and, thus, ensue recovery from subclinical mastitis in the Indian water buffaloes.     

Assessment of Detoxifying Markers for Florfenicol in Rainbow Trout Liver

Florfenicol (FF) is employed in fish farms to contest or prevent bacterial infections. However, this pharmaceutical may produce reactive oxygen species that may cause biochemical changes in antibiotic-treated fish. The aim of this study was to evaluate the effect of FF on Rainbow Trout Oncorhynchus mykiss treated for 10 d with 7.5 and 15 mg/kg FF followed by a withdrawal period of 5 d. Superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, glyoxalase I and glyoxalase II, total glutathione, lactic dehydrogenase, and alkaline phosphatase were investigated in the livers of treated and untreated fish. A general impairment of antioxidant enzymes and metabolic indicators was measured in FF-treated Rainbow Trout. Onset of oxidative damage may have occurred during the antibiotic treatment as a consequence of the effect of FF toxicity at mainly the highest dose. Nevertheless, the rise in levels of total glutathione and glutathione S-transferase even after the withdrawal period may shield the antibiotic-mediated oxidative processes.

Received December 22, 2015; accepted May 26, 2016 Published online October 28, 2016     

Ni和Cu胁迫对骆驼蓬抗氧化酶活性的影响

采用温室盆栽法,研究Ni、Cu单一处理对金昌Ni/Cu尾矿库先锋植物骆驼蓬幼苗Ni、Cu积累、膜脂过氧化程度与抗氧化酶系统的影响。结果表明,骆驼蓬幼苗叶片及根部Ni、Cu含量随土壤介质中Ni、Cu含量的增加而增加,Ni主要富集在骆驼蓬的叶片,而Cu主要富集在骆驼蓬的根部;叶片及根部丙二醛(MDA)含量总体随Ni、Cu处理浓度升高而增加,膜脂过氧化程度加剧;Ni胁迫下,骆驼蓬叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸酶(APX)和谷胱甘肽还原酶(GR)活性较对照均有所增强,共同组成植物体内一个有效的活性氧自由基(ROS)清除系统,根部SOD、APX和CAT活性受到抑制,POD和GR受诱导活性增强,在减轻Ni胁迫引起氧化损伤方面起重要作用;Cu胁迫下骆驼蓬叶片SOD、POD、CAT、APX及GR活性较对照均有所提高,减弱了Cu胁迫引起的氧化损伤;根部APX活性受抑制,SOD、POD、CAT及GR活性受诱导增强,在缓解Cu胁迫引起氧化损伤方面发挥重要作用。     

Effect of dietary microbially produced gamma‐linolenic acid and plant extracts on enzymatic and non‐enzymatic antioxidants in various broiler chicken organs

Plant extracts and fungal fermented feed with gamma‐linolenic acid‐rich microbial oils are perspective additives for use in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for the prevention and treatment for certain pathological conditions, and as antioxidants. The activity of antioxidant enzymes and the level of reduced glutathione were measured in the plasma and in liver, heart and kidney mitochondria after 42 days of feeding broiler chickens both regular and combination diets. These were selected based on our previous experience. The administration of agrimony and gamma‐linolenic acid resulted in a significant decrease in superoxide dismutase activity in all four bodies in contrast to plant extracts. We conclude that the decrease in activity is due to decreased production, and hence dismutation, of superoxide radicals to peroxides followed by lower activity of glutathione peroxidase, which was not seen in the case of only plant extract administration. Generally, higher glutathione reductase activity would be in response to increased demands on reduced glutathione as a cofactor for the reaction catalysed by glutathione peroxidase and the utilization of glutathione itself. However, measured levels of reduced glutathione showed no change. The results argue against any oxidative stress conditions. The application of agrimony extract appears to be suitable for the antioxidant effect against peroxidation of gamma‐linolenic acid. As the efficacy of measuring the effects of diets on the oxidative stability of meat caused by selected antioxidant enzymes is rather low, additional data from the experiment will be processed to clearly assess the influence of this combination of diets.