Ethanol- and nicotine-induced membrane changes in embryonic and neonatal chick brains

In order to study the effects of EtOH and/or nicotine on brain membrane fatty acid composition, various concentrations of EtOH and/or nicotine were injected into the air sac of chicken eggs at 0 days of incubation. Controls were injected with saline. Experimental groups were injected with either 200 micromol EtOH/kg egg, 100 micromol nicotine/kg egg, 200 micromol nicotine/kg egg, 200 micromol EtOH/kg and 100 micromol nicotine/kg egg, or 200 micromol EtOH/kg and 200 micromol nicotine/kg egg. In all experimental groups, EtOH- and nicotine-induced decreases in brain long-chain polyunsaturated membrane fatty acids were observed in stage 44 embryos, stage 45 embryos, and neonatal chicks. These EtOH- and nicotine-induced decreases in brain membrane polyunsaturated fatty acids correlated with elevated levels of brain lipid hydroperoxides and reduced brain acetylcholinesterase (AChE; EC. 3.1.1.7) activities.     

Effect of indole acetic acid administration on the neutrophil functions and oxidative stress from neutrophil, mesenteric lymph node and liver

This study was done to investigate the effect of the in vivo administration of indole acetic acid (IAA) on the neutrophil function, the activities of antioxidants enzymes in neutrophils, the mesenteric lymph node and on the oxidative stress in liver and plasma. The animals received subcutaneous administration of IAA in a phosphate-buffered saline (the control group received only the phosphate-buffered saline). The other groups received IAA at concentrations of 1 mg (T1), 2 mg (T2) and 18 mg (T3) per kg of body mass per day. Administration of IAA in both treatments T2 and T3 promoted a significant rise in the phagocytic capacity of neutrophils (by 51%), in comparison with the control. Another alteration was observed in antioxidant enzyme activities of the neutrophil and lymph node. But in the liver, the treatments imposed a significant decrease in the activity of catalase of 19% and 30% for T2 and T3, respectively, in comparison with the control. A similar effect was observed in the activity of hepatic glutathione peroxidase for T3 where a significant decrease of 31%, compared with the control, was obtained. The IAA did not show another significant alteration of the activities of superoxide dismutase and glutathione reductase activities in liver. The hepatic lipid peroxidation level, available by reactive products with thiobarbituric acid, has shown a significant decrease of 27% and 29% with T1 and T3 respectively, in comparison with the control. IAA treatment did not show a significant alteration in reduced glutathione contents in comparison with the liver and plasma controls. In conclusion, the IAA administration has a good potential animal utilization for increasing the phagocytic capacity with no prooxidant effect.     

重组酿酒酵母合成谷胱甘肽的研究

利用PCR方法以酿酒酵母Saccharomyces cerevisiae S288c基因组DNA为模板;PCR扩增γ -谷氨酰半胱氨酸合成酶(GSH1)及谷胱甘肽合成酶(GSH2)基因;以乙醇脱氢酶(ADH1)启动子进行表达;并以Kan^r基因和Hyg^r基因作为筛选标记;构建了两个重组表达质粒YEplace181AK-GSH1和YEplace181AH-GSH2。采用电击法将这两个质粒分别和同时转入工业酿酒酵母菌株Saccharomyces cerevisiae TS013;在含有G418或（和）Hygromycin的平板上筛选阳性克隆S.TS013/GSH1、S.TS013/GSH2和S.TS013/GSH1+GSH2。重组菌进行摇瓶发酵;采用四氧嘧啶法测定培养细胞合成谷胱甘肽含量。结果显示;不同转化子重组菌的胞内谷胱甘肽产量比宿主菌分别提高了44.11%、29.79%和56.47%。     

The effect of growth medium on the antioxidant defense of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

We compared the oxidation of dihydrorhodamine 123, glutathione contents and activities of superoxide dismutase (SOD) and catalase for three wild-type strains of Saccharomyces cerevisiae grown on media with different carbon sources. The rate of oxidation of dihydrorhodamine 123 was much higher in respiring cells grown on ethanol or glycerol media than in fermenting cells grown on glucose medium. The total SOD activity was highest on glycerol medium and lowest on ethanol medium, while the catalase activity was highest on glycerol medium. The sequence of glutathione content values was: glucose > ethanol > glycerol.     

Superoxide dismutase and glutathione reductase overexpression in wheat protoplast: photooxidative stress tolerance and changes in cellular redox state

In previous works, we have established a correlation between antioxidant system response and tolerance to drought, osmotic stress and photooxidative stress of different wheat cultivars with contrasting drought tolerance. In the present work, a protocol to obtain and transform wheat protoplasts was established. Transgenic protoplasts with Manganese Superoxide Dismutase (Mn-SOD) (E.C.: 1.15.1.1) and Glutathione Reductase (GR) (E.C.: 1.6.4.2) overexpression in chloroplasts were obtained, and their responses to photooxidative stress were characterized. Protoplasts with Mn-SOD or GR overexpression, showed different responses and tolerance to photooxidative stress. Protoplasts with Mn-SOD overexpression showed lower levels of oxidative damage, higher level of endogenous hydrogen peroxide and a great induction of total SOD and GR activities during photooxidative treatments. In protoplasts with GR overexpression the oxidative damage provoked by the photooxidative treatment was similar to control protoplasts, the GSH content and GSH/GSH + GSSG ratio were higher than control and Mn-SOD transformed protoplast, and total SOD and GR activities were not induced. Our results suggest that the differential responses and tolerance to photooxidative stress given by Mn-SOD or GR overexpression, also depend on the effects of these enzyme activities over the cellular redox state balance, which modulate the responses to photooxidative stress.     

麦角新碱联合米索前列醇治疗前置胎盘的临床疗效及对血清ROS、SOD、GSH-px水平的影响

目的:分析麦角新碱联合米索前列醇治疗前置胎盘的临床疗效及对血清活性氧(ROS)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-px)水平的影响。方法:选择2016年5月-2017年5月我院收治的前置胎盘患者100例纳入本次研究,根据随机分组法分为观察组(n=52)和对照组(n=48)。对照组使用米索前列醇治疗,观察组采用麦角新碱联合米索前列醇进行治疗。比较两组患者的临床疗效、治疗前后血清ROS、SOD、GSH-px水平的变化、术中、术后出血量、止血时间及不良反应的发生情况。结果:治疗后,两组患者的总有效率分别为96.15%、81.25%,观察组显著高于对照组(P0.05);两组患者血清ROS、SOD、GSH-px水平均较治疗前显著改善,且观察组患者血清ROS低于对照组,血清SOD、GSH-px水平显著高于对照组(P0.05);观察组患者术中、术后出血量及止血时间均显著低于对照组(P0.05);两组患者不良反应总发生率分别为7.69%、22.92%,观察组显著低于对照组(P0.05)。结论:麦角新碱联合米索前列醇治疗前置胎盘的疗效和安全性均显著优于单用米索前列醇治疗,可能与其有效改善血清ROS、SOD、GSH-px水平有关。     

PprI和RecX蛋白对耐辐射奇球菌抗氧化作用的影响

利用基因突变、化学发光法和酶活性分析研究了耐辐射奇球菌中与辐射抗性密切相关的基因pprI(Dr0167)和recX(Dr1310)突变对菌体活性氧清除作用的影响,分析了其对抗氧化酶活性的调控功能。实验结果表明,缺失pprI的突变株对活性氧自由基氧化异常敏感,过氧化氢酶和超氧化物歧化酶活性显著降低。与之相反,RecX对菌体活性氧清除作用表现为一种“负”的影响,即缺失recX的突变株对活性氧自由基的清除能力反而增强了,过氧化氢酶和超氧化物歧化酶的酶活性明显增加。表明这两个基因与抗氧化系统的调控有关。为进一步研究该菌的抗氧化机制提供了一些思路。     

Regulation of phytochelatin synthesis by zinc and cadmium in marine green alga,Dunaliella tertiolecta

Although Cd(2+) is a more effective inducer of phytochelatin (PC) synthesis than Zn(2+) in higher plants, we have observed greater induction of PC synthesis by Zn(2+) than Cd(2+) in the marine green alga, Dunaliella tertiolecta. To elucidate this unique regulation of PC synthesis by Zn(2+), we investigated the effects of Zn(2+) and Cd(2+) on the activities of both phytochelatin synthase (PC synthase) and enzymes in the GSH biosynthetic pathway. PC synthase was more strongly activated by Cd(2+) than by Zn(2+), but the difference was not very big. On the other hand, gamma-glutamylcysteine synthetase (gamma-ECS) and glutathione synthetase (GS) were activated by both heavy metals, but their activities were higher in Zn-treated cells than in Cd-treated cells. Dose-dependent stimulation of intracellular reactive oxygen species (ROS) production was observed with Zn(2+), but not Cd(2+) treatment. These results suggest that Zn(2+) strongly promotes the synthesis of GSH through indirect activation of gamma-ECS and GS by stimulating ROS generation. This acceleration of the flux rate for GSH synthesis might mainly contribute to high level PC synthesis.     

A spectrophotometric assay of gamma-glutamylcysteine synthetase and glutathione synthetase in crude extracts from tissues and cultured mammalian cells

An assay of gamma-glutamylcysteine synthetase (gamma-GCS) and glutathione synthetase (GS) in crude extracts of cultured cells and tissues is described. It represents a novel combination of known methods, and is based on the formation of glutathione (GSH) from cysteine, glutamate and glycine in the presence of rat kidney GS for the assay of gamma-GCS, or from gamma-glutamylcysteine and glycine for the assay of GS. GSH is then quantified by the Tietze recycling method. Assay mixtures contain the gamma-glutamyl transpeptidase (GGT) inhibitor acivicin in order to prevent the degradation of gamma-glutamylcysteine and of the accumulating GSH, and dithiothreitol in order to prevent the oxidation of cysteine and gamma-glutamylcysteine. gamma-GCS and GS levels determined by this method are comparable to those determined by others. The method is suitable for the rapid determination of gamma-GCS GS in GGT-containing tissues and for the studies of induction of gamma-GCS and GS in tissue cultures.     

PEGylated ceria nanoparticles used for radioprotection on human liver cells under γ-ray irradiation

Ceria nanoparticles (CNPs) have recently been shown to protect cells and animals from radiation-induced damage. However, most of the CNPs used in previous studies were either naked or weakly protected by surfactants, which inevitably encounter many obstacles in biological applications. Here, alendronate was used as an ideal anchor to graft polyethylene glycol (PEG) onto CNPs, leading to enhanced stability, reduced cytotoxicity, and improved biological properties. Further investigation assessed the protective ability of the nanoparticles against radiation-induced effects for human normal liver cells (L-02), indicating that the PEGylated CNPs (CNPs–AL–PEG) were more efficient than naked CNPs. We determined that enhanced Ce³⁺/Ce⁴⁺ ratios improved intracellular dispersion and that the ameliorated intracellular distribution of CNPs–AL–PEG contributes to the elevated expression of SOD2, which leads to increased protection of normal cells against ROS and reduces the oxidatively generated DNA damage. These studies hold tremendous promise for radioprotection and biological applications.     

不同剂量白藜芦醇对糖尿病性白内障大鼠晶状体抗氧化酶活力的影响

目的:探究不同剂量白藜芦醇对糖尿病性白内障大鼠晶状体抗氧化酶活力的影响。方法:75只5周龄健康SPF级雄性SD大鼠按照随机数字表法分为正常对照组、模型组,白藜芦醇低剂量组,白藜芦醇中剂量组和白藜芦醇高剂量组,每组各15只。五组大鼠均给予常规适应性喂养,模型组和白藜芦醇低、中、高剂量组大鼠采用链脲佐菌素(STZ)以60 mg/kg的给药剂量制作糖尿病大鼠模型,成模后白藜芦醇低剂量组按20 mg/kg、白藜芦醇中剂量组按50 mg/kg、白藜芦醇高剂量组按100 mg/kg的给药剂量每日给予白藜芦醇灌胃。观察12周后5组大鼠晶状体的混浊程度,检测血糖、体重后处死大鼠,检测晶状体内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)以及过氧化氢酶(CAT)的活性改变。结果:随着白藜芦醇剂量的升高,白藜芦醇低、中、高剂量组大鼠的血糖逐渐降低而体重逐渐升高,且组间比较均具有统计学差异(P0.05)。三组不同剂量白藜芦醇干预组大鼠晶状体的浑浊程度均低于模型组,且白藜芦醇高剂量组大鼠晶状体的浑浊程度最低,差异均具有统计学意义(P0.05)。白藜芦醇低、中、高剂量组大鼠SOD、GSH-PX和CAT酶活力与模型组大鼠相比均明显升高,而与正常对照组相比均明显降低(均P0.05)。随着白藜芦醇剂量的升高,白藜芦醇低、中、高剂量组大鼠SOD、GSH-PX和CAT酶活力逐渐升高,且组间比较均具有统计学差异(P0.05)。结论:高剂量白藜芦醇可更为明显地降低血糖浓度,提高晶状体SOD、GSH-Px及CAT酶活力,改善糖尿病性白内障晶状体的浑浊程度。     

Amelioration of glucose induced hemolysis of human erythrocytes by vitamin E

Cells under aerobic condition are always threatened with the insult of reactive oxygen species, which are efficiently taken care of by the highly powerful antioxidant systems of the cell. The erythrocytes (RBCs) are constantly exposed to oxygen and oxidative stress but their metabolic activity is capable of reversing the injury under normal conditions. In vitro hemolysis of RBCs induced by 5, 10 and 20 mM glucose was used as a model to study the free radical induced damage of biological membranes in hyperglycemic conditions and the protection rendered by vitamin E on the same. RBCs are susceptible to oxidative damage, peroxidation of the membrane lipids, release of hemoglobin (hemolysis) and alteration in activity of antioxidant enzymes catalase and superoxide dismutase. The glucose induced oxidative stress and the protective effect of vitamin E on cellular membrane of human RBCs manifested as inhibition of membrane peroxidation and protein oxidation and restoration of activities of superoxide dismutase and catalase, was investigated.Thiobarbituric acid reactive substances are generated from decomposition of lipid peroxides and their determination gives a reliable estimate of the amount of lipid peroxides present in the membrane. Vitamin E at 18 μg/ml (normal serum level) strongly enhanced the RBC resistance to oxidative lysis leading to only 50–55% hemolysis in 24 h, whereas RBCs treated with 10 and 20 mM glucose without vitamin E leads to 70–80% hemolysis in 24 h. Levels of enzymic antioxidants catalase, superoxide dismutase and nonenzymic antioxidants glutathione showed restoration to normal levels in presence of vitamin E. The study shows that vitamin E can protect the erythrocyte membrane exposed to hyperglycemic conditions and so a superior antioxidant status of a diabetic patient may be helpful in retarding the progressive tissue damage seen in chronic diabetic patients.     

大豆萌发过程的活性氧代谢

本文研究了大豆萌发过程中活性氧的产生与清除,并探讨了光因子在活性氧代谢中的作用。大豆呼吸强度、O产生速率及H2O2水平都在吸水后第四天达到高峰,然后下降,三者的变化趋势同步。SOD、POD及APX的活性随萌发过程而逐渐增强,最后趋于平稳。SOD同工酶谱中分别于萌发的第二、第三天各出现一条新的酶带。CAT在萌发的初期猛增50倍左右,之后趋于稳定。在三种清除H2O2的酶（CAT、POD、APX）中,CAT清除H2O2的能力远远高于POD与APX,CAT可能是大豆萌发过程中最主要的H2O2清除酶。光萌发时呼吸强度低于暗中萌发,但O产生速率与H2O2水平高于暗萌发,光萌发时O的产生占总耗氧量的1．1—2．7％,而暗中萌发为0．9—1．3％。光条件下SOD、APX活性明显高于暗中萌发,而POD与CAT则在光和暗条件下相差不大。     

Pair-wise linear and 3D nonlinear relationships between the liver antioxidant enzyme activities and the rate of body oxygen consumption in mice

Relationships between the rate of body oxygen consumption (VO₂) and the liver key antioxidant enzyme activities were assessed in female CBA mice. The pair-wise linear regression and correlation demonstrated significant correlative links between VO₂ and activity of catalase (CAT). Nonlinear 3D plotting revealed a complementary pattern of CAT and glutathione peroxidase (GP) relation. CAT activity was elevated in mice with proportionally high VO₂ and superoxide dismutase (SOD), whereas GP activity was high in animals with low or disproportional VO₂ and SOD.     

A polysaccharide–peptide complex from abalone mushroom (Pleurotus abalonus) fruiting bodies increases activities and gene expression of antioxidant enzymes and reduces lipid peroxidation in senescence-accelerated mice

The antioxidant effects of a polysaccharide–peptide complex (F22) from mushroom (Pleurotus abalonus)-fruiting bodies were studied. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in the liver, kidney, and brain of senescence-accelerated mice showed a marked increase after treatment with the polysaccharide–peptide complex. Concurrently, the gene expression levels of SOD, CAT, and GPx, as determined with real-time polymerase chain reaction, were up-regulated in the liver, kidney, and brain, whereas the MDA content in these organs declined. The maximal lifespan of the mice was prolonged.     

Alterations in Protective Enzymes Against Peroxidation in the Central and Peripheral Nervous System of Control and Dysmyelinating Mutant Mice

The activities of peroxide-detoxifying enzymes such as superoxide dismutase (SOD), glutathione peroxidase, glutathione reductase, and catalase were measured in the nervous system of neurological dysmyelinating mutants: quaking (Qk), shiverer (Shi), and trembler (Tr) mice. Cu/Zn-SOD activity was higher in the cerebellum of Qk and Shi mice (by 53% and 106%, respectively) in comparison with controls, but it was the same in the cerebellum of Tr mice and their corresponding controls. In contrast, there was no difference in the level of Cu/Zn-SOD activity in the cerebrum of Qk, Shi, and Tr mice and their respective controls. Mn-SOD activity was the same among all the mutants compared to control animals in both cerebrum and cerebellum. In Shi cerebellum, glutathione peroxidase and glutathione reductase activities were slightly decreased (a 21.6% and a 13.2% diminution, respectively), whereas catalase activity in cerebrum and cerebellum was the same among mutants and control mice. In the sciatic nerve from Tr mice, all the enzymatic activities were enhanced: sixfold increase for total SOD, and 2.4-fold, 3.5-fold, and 1.8-fold increase for glutathione peroxidase, glutathione reductase, and catalase, respectively.