Antioxidant processes are affected in juvenile rainbow trout (Oncorhynchus mykiss) exposed to ozone and oxygen-supersaturated water

Two groups of juvenile rainbow trout (Oncorhynchus mykiss) were exposed for 4 h to either hyperoxic (>40 mg O₂/l) or ozonised (5.2 μg O₃/l) water containing supersaturated oxygen. Gill, liver, blood cells and plasma were taken 1, 5, 12, 24 and 48 h following the exposure periods and antioxidant responses and oxidative damage studied in terms of, respectively, antioxidant enzyme activities/glutathione status and lipid peroxidation. Observed biochemical changes were significant at p<0.05. Ozonation elevated oxidised glutathione (GSSG) levels, increased lipid peroxidation and decreased reduced glutathione (GSH) levels in gills 1 h following exposure, which indicate oxidative stress. However, the longer-term effects (48 h) following ozone exposure resulted in increased GSH levels in both gills and liver and, measured as oxidative stress index (OSI), were indicative of enhanced potential of tissues to resist oxidative stress. Ozonation also resulted in elevated activity of superoxide dismutase (SOD) at 5–12 h following exposure in gills compared to 24–48 h in liver, indicating either differential regulation and time-courses of response, or an earlier impact of ozonolysis products on gills than liver. In contrast, catalase activity was elevated in both gills and liver at 24 h by the effects of hyperoxia alone. Hyperoxia also decreased GSSG levels in both gills and liver, but had no effect on lipid peroxidation. Increases were seen in the activities of selenium-dependent glutathione peroxidase as a result of ozonation (gills; 5 and 24 h) and hyperoxia (liver; 1 h), and in total GPX (Se-dependent and independent) as a result of ozonation (both tissues; 48 h), but no effects were seen on glutathione reductase (GR) in either tissues. Levels of GSH were increased in blood cells at various times following hyperoxia. Overall, the results show (i) differential antioxidant and pro-oxidant effects of ozonation compared to hyperoxia, (ii) gills and possibly blood as the first line of impact and defence, with later effects on liver, and (iii) that even after 24–48 h following ozone exposure, all antioxidant defences had not returned to pre-exposure values.     

Effect of 7,8-Dihydroxyflavone as an Antioxidant on In Vitro Maturation of Oocytes and Development of Parthenogenetic Embryos in Pigs

One of the factors that impairs in vitro produced porcine embryosis the oxidative stress that is mainly caused by the imbalance between reactiveoxygen species (ROS) generation and antioxidants activity, especially that ofglutathione (GSH). Here, we examined the effect of 7,8-dihydroxyflavone (7,8-DHF), akind of flavonoid antioxidant, on porcine oocyte maturation and its developmentalcompetence. Porcine oocytes were cultured in media supplemented with 0, 1, 5 and 10μM 7,8-DHF during both in vitro maturation (IVM) and invitro culture (IVC) after parthenogenetic activation. Maturation ofoocytes was evaluated based on first polar body (PB) extrusion and intracellular GSHlevel, and developmental competence was assessed through observing cleavage andblastocyst formation. In each step, the levels of intracellular GSH and ROS wereassessed by fluorescence intensity, and the apoptosis-related gene expression wasexamined using semiquantitative RT-PCR. The group treated with 1 μM 7,8-DHF duringIVM and IVC showed increased cytoplasmic maturation and reached the blastocysts stage(36.1%) at a higher rate than the other groups (24.7, 16.0 and 10.3% for 0, 5 and 10μM, P<0.05). In that group, the intracellular GSH level was significantlyincreased while ROS generation was significantly decreased after IVM and IVC(P<0.05). Moreover, it showed high expression of an anti-apoptotic gene(BCL2L1) and low expression of a pro-apoptotic gene(BAK1) (P<0.05). In conclusion, treatment with 1 μM 7,8-DHFduring IVM and IVC showed an anti-apoptotic effect by increasing intracellular GSHsynthesis and scavenging ROS and therefore improved the developmental competence ofporcine embryos.     

氮素形态对烤烟成熟过程中抗坏血酸-谷胱甘肽循环的影响

探讨了氮素形态对烤烟成熟过程中抗坏血酸-谷胱甘肽循环系统的影响。结果表明,烟叶自移栽后60 d开始,抗坏血酸过氧化物酶(APX)活性和抗坏血酸(ASA)含量逐渐上升,到70~80 d达到最高峰,随后稳定在一定的水平。3种不同氮素形态处理的APX活性和ASA含量变化规律相似,均随着硝态氮比例的增加而增大;谷胱甘肽还原酶(GR)活性在移栽后90 d达最大值,此后逐渐下降,谷胱甘肽(GSH)含量随着烟叶的成熟持续下降。烟叶成熟过程中各处理GR活性和GSH含量随着铵态氮比例的增加而增大。     

Antioxidant enzymes and somatic cell count in dairy cows fed with organic source of zinc,copper and selenium

The objectives of this study were to evaluate the effects of dietary organic sources of zinc (Zn), copper (Cu) and selenium (Se) for dairy cows on somatic cell count (SCC), occurrence of subclinical mastitis, number of clinical mastitis cases and concentration of superoxide dismutase (CuZnSOD), glutathione peroxidase (GSH-Px) and ceruloplasmin (CP). Nineteen dairy cows, with confirmed pregnancy, were selected by body weight, body condition score (BCS), number of lactation, and milk yield in previous lactation, and randomly distributed among two groups, one to receive organic (n = 9) and the other the inorganic (n = 10) source of Zn, Cu and Se. Diets were formulated to meet the nutritional requirements of animals from 60 days before the expected date of calving up to 80 days of lactation. Blood samples were collected at − 60, − 21, 1, 21, 40 and 80 days in relation to the expected date of calving for analysis of CuZnSOD, GSH-Px and CP concentration. Milk samples for SCC determination were collected weekly after 15 days of lactation, also at day 1 and 7 and when a clinical case was diagnosed for microbiological culture. The number of new and total cases of subclinical mastitis was lower for the group of cows fed with organic sources of Zn, Cu and Se compared to animals receiving inorganic sources. Average SCC during the first 80 days of lactation tended to be lower for the group fed with organic Zn, Cu and Se. Feeding organic Zn, Cu and Se did not alter the concentration of CuZnSOD, GSH-Px and CP.     

Inhibitory effect of iron on vasodilatation in the isolated rat aorta

AIM: The objectives of the present study were to examine the effect of iron on relaxation of isolated rat aortic rings, and to elucidate the underlying mechanism. METHODS: The thoracic aortic rings of male Sprague-Dawley rats were mounted on bath system. Vasodilatation of aortic rings preconstricted with 10^-6 mol/L of phenylephrine (PE) was measured. RESULTS: (1) Exposure of endothelium-intact aortic rings to ferric ammonium citrate (FAC) for 30 min caused a significant reduction in the relaxation response to acetylcholine (ACh). Pretreatment with L-arginine (L-Arg) before incubation with FAC did not reverse the inhibition of relaxation response to ACh completely. (2) In endothelium-intact aortic rings, L-Arg relaxed the PE preconstricted vessels. Exposure to FAC for 30 min caused the decrease in the relaxation response to L-Arg. There was no difference in the relaxation response to nitric oxide donor, sodium nitroprusside, between endothelium-denuded arteries treated with or without FAC. (3) Dimethyl sulfoxide had no effect on the inhibition of relaxation to ACh by FAC in endothelium-intact rings. Pretreatment of arteries with glutathione and catalase prevented the decrease in relaxation responses to ACh induced by FAC. (4) The nitric oxide synthase activity was (56.49±2.49)×10³U/g protein in normal aorta with endothelium, while after incubation with FAC for 30 min, it reduced to (25.15±5.75)×10³U/g protein ( P< 0.05). CONCLUSION: Inactivation of nitric oxide synthase and decrease in intracellular glutathione level might mediate iron-induced inhibition of arterial relaxation responses to ACh.     

谷胱甘肽S-转移酶综述

谷胱甘肽S-转移酶(glutathioneS-transferases,GSTs)是由多个基因编码、具有多种功能的超基因家族酶,是多种生物体内的主要解毒系统。本文综述了GSTs的分型、结构等方面的研究进展。     

Effects of carbaryl and azinphos methyl on juvenile rainbow trout (Oncorhynchus mykiss) detoxifying enzymes

In this study, the effects of sublethal exposures to the anticholinesterase insecticides azinphos methyl (AzMe) and carbaryl on the detoxifying responses of juvenile rainbow trout Oncorhynchus mykiss were investigated. Juvenile specimen were exposed to sublethal concentrations of AzMe (2.5 and 5 μg/L) and carbaryl (1 and 3 mg/L) for 24, 48 and 96 h. Carboxylesterase (CbE), catalase (CAT) and glutathione S-transferase (GST) activities as well as reduced glutathione (GSH) and cytochrome P450-1A (CYP1A) levels were monitored in liver and/or kidney. In all exposed groups liver CbE was significantly inhibited. Liver and kidney GSH level was reduced after sublethal exposure to both compounds. Carbaryl induced CAT activity during the first 48 h of exposure, followed by a significant decrease, whereas AzMe continuously decreased CAT activity. GST activity and CYP1A were transiently induced at 24 h by carbaryl exposure (3 mg/L) but sublethal exposure to AzMe did not affect GST activity or CYP1A. Our results show that the O. mykiss detoxifying system are a target for carbaryl and AzMe action, probably affecting redox balance. Although the responses showed similar trends in both organs, they were more important in liver than in kidney. The early inhibitory effect in CAT activity and GSH content produced by AzMe may be associated with a high degree of oxidative stress. Early induction of CYP1A, GST and CAT by carbaryl followed by enzyme inhibition suggests a milder or delayed oxidative stress, revealing differences between both pesticides metabolization. CbE inhibition is a good biomarker for AzMe and carbaryl exposure.     

槲皮素对棉铃虫体内一些解毒酶系和靶标酶的诱导作用

用培养基混药法研究了槲皮素对棉铃虫羧酸酯酶、谷胱甘肽转移酶的诱导作用和对乙酰胆碱酯酶敏感度的影响。槲皮素诱导种群对甲基对硫磷的敏感度明显降低 ,对灭多威和溴氰菊酯的敏感度没有明显变化。诱导种群羧酸酯酶的活性提高 2～ 3倍 ,谷胱甘肽转移酶活性提高近 15倍 ,而乙酰胆碱酯酶对对氧磷的敏感度却提高了 3.5倍。说明槲皮素诱导种群对甲基对硫磷的敏感度降低是由于解毒酶系和乙酰胆碱酯酶综合作用的结果。     

谷胱甘肽对盐胁迫下玉米幼苗抗氧化特性和光合性能的影响

以80mmol/L NaCl模拟盐胁迫,研究谷胱甘肽(GSH)对盐胁迫下‘滑玉14’幼苗叶片抗氧化特性和光合性能的影响。结果表明:盐胁迫显著提高叶片细胞质膜透性、丙二醛(MDA)质量摩尔浓度、抗氧化酶超氧化物歧化酶(SOD)、抗坏血酸氧化酶(APX)和脱氢抗坏血酸还原酶(DHAR)活性、抗氧化物质谷胱甘肽(GSH)和抗坏血酸(AsA)质量摩尔浓度及叶绿素荧光参数非光化学猝灭系数(qN),显著降低叶绿素荧光参数PSⅡ最大光化学效率(F_v/F_m)、光化学猝灭系数(qP)和PSⅡ实际量子产额(Φ_(PSⅡ)),光合色素质量分数、光合速率和单株生物量干质量。这说明,盐胁迫对‘滑玉14’造成氧化胁迫,并对光系统Ⅱ的功能造成伤害。50mg/L GSH处理则可以显著提高盐胁迫下玉米幼苗叶片过氧化物酶(POD)、SOD、谷胱甘肽还原酶(GR)活性、GSH质量摩尔浓度、F_v/F_m、qP和Φ_(PSⅡ)、光合色素质量分数、光合速率和单株生物量干质量,并显著降低细胞质膜透性、MDA质量摩尔浓度和qN。上述研究结果说明,GSH处理可以提高玉米幼苗的抗氧化能力和光合性能,从而缓解盐胁迫造成的伤害。     

山羊草谷胱甘肽S-转移酶基因家族鉴定及表达分析

谷胱甘肽S-转移酶是一种多功能蛋白酶,在植物体内参与干旱、盐、低温、重金属等多种非生物胁迫的调节;山羊草是普通小麦D染色体组的供体物种,深入挖掘山羊草中GST基因,对进一步分析六倍体小麦GST基因的功能具有重要意义。本研究利用信息生物学手段,在山羊草中共发现114条GST基因序列,分属于6个亚族;基因复制分析发现共4对基因发生了基因复制,且均为纯化选择;采用荧光定量PCR对部分GST基因在非生物胁迫下的表达分析发现,8个GST基因在响应干旱和盐胁迫时,主要在根部显著上调表达,3个GST基因在响应低温胁迫时,在根和叶中均显著上调表达,说明山羊草中的GST基因在应答非生物胁迫时,在不同组织中的表达存在着差异。