组织中氧化型和还原型谷胱甘肽荧光测定法

介绍了一种同时测定组织中氧化型谷胱甘肽(GSSG)和还原型谷胱甘肽(GSH)的荧光方法,应用邻苯二甲醛作为荧光试剂,GSH 和 GSSG 的标准曲线(2—10μg)均呈线性关系.测定了17例正常人肺组织及肺癌组织胞浆和线粒体内GSH 含量的亚细胞分布,本法简单、灵敏、重复性好、回收完全,比高效液相层析法容易推广.     

亚硒酸钠对小麦幼苗中谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性以及谷胱甘肽含量的影响

用1.0 mg·L-1的亚硒酸钠根施小麦幼苗,测定亚硒酸钠对谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性以及还原性谷胱甘肽含量的结果表明,外源亚硒酸钠对麦苗地上部的谷胱甘肽过氧化物酶和谷胱甘肽转硫酶活性均有诱导作用,使麦苗体内的谷胱甘肽含量水平增加.     

水分胁迫对刺槐叶和根谷胱甘肽抗氧化系统的影响

在人工控水条件下,采用土壤最大持水量70%、55%、40%的水分处理模拟环境中的正常水分、轻度和重度水分胁迫处理,测定了刺槐叶片和根系中还原型谷胱甘肽(GSH)和还原型抗坏血酸(AsA)含量以及谷胱甘肽还原酶(GR)、谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性,以探讨水分胁迫条件下刺槐谷胱甘肽抗氧化系统的保护作用.结果显示:各水分处理的刺槐叶片GSH和AsA含量及GR 和SOD活性均明显高于根,根中GSH-Px活性只有在重度水分胁迫处理下大于叶片.随水分胁迫加剧,刺槐GSH含量在叶片中先升高后降低,在根中不断升高;AsA含量在叶中持续降低,在根中先升高后降低;GR活性在叶片和根系中都会降低,GSH-Px和SOD活性在叶中先升高后降低,在根中均持续升高.研究表明,刺槐谷胱甘肽抗氧化系统的GSH和GSH-Px对干旱胁迫诱发的活性氧清除起主要作用,同时提高GSH含量和GSH-Px活性是刺槐应对干旱胁迫的重要措施.     

谷胱甘肽的研究与应用

谷胱甘肽的研究与应用刘振玉（天津体育学院基础部,天津３００３８１）关键词还原型谷胱甘肽,氧化型谷胱甘肽谷胱甘肽（ＧＳＨ）是由谷氨酸、半胱氨酸和甘氨酸形成的三肽化合物,在生物体内有许多重要作用。合成谷胱甘肽的第一步是在谷氨酸的γ－羧基与半胱氨酸的氨基之...     

谷胱甘肽对小麦幼穗胚性愈伤组织形成以及几种相关酶活性的影响

用谷胱甘肽（GSH）处理小麦幼穗,其愈伤组织绿点率明显高于对照和BSO（buthionine sulfoximine)处理,这种效应与后期GSH诱导谷胱甘肽转移酶（GST）和醛氧化酶（ALD）活性的增加一致。     

S-谷胱甘肽化修饰的研究进展

S-谷胱甘肽化(S-glutathionylation)是谷胱甘肽和靶蛋白半胱氨酸残基之间形成混合二硫化物的过程.由于其能调节靶蛋白功能,因此也属于蛋白质翻译后修饰.与其相对应,蛋白质的去谷胱甘肽化可由谷氧还蛋白(Grx)催化.因此,S-谷胱甘肽化修饰也被认为是一种防止蛋白质半胱氨酸巯基发生不可逆修饰的保护机制.由于该修饰还会改变含有巯基的氧化还原敏感型蛋白的结构与功能,因此也属于蛋白质功能调节的重要方式.哺乳动物细胞中S-谷胱甘肽化水平的改变与许多病理机制有关,但S-谷胱甘肽化在植物中的研究还处于起步阶段.本文综述了蛋白质的S-谷胱甘肽化的反应机制、检测方法、生理作用的相关研究进展,最后还提出今后研究中要解决的重要问题.     

谷胱甘肽化修饰与氧化还原信号转导

蛋白质谷胱甘肽化（S-glutathionylation）是一种重要的翻译后修饰方式,氧化还原信号转导途径的很多相关分子都可受到谷胱甘肽化的调节,尤其是一些重要的蛋白激酶和转录因子。因此蛋白质的谷胱甘肽化修饰日益引起人们的重视。人们推测,谷胱甘肽化可能是细胞内氧化还原信号转导的一种重要机制。     

发酵法生产谷胱甘肽的研究进展

谷胱甘肽是一种含巯基活性三肽。简要综述发酵法生产谷胱甘肽中的菌种选育、发酵调控策略、提取及分离纯化等方面的研究进展。     

关于巯基和Mn~(2+)介导豆壳过氧化物酶氧化藜芦醇的研究

藜芦醇作为非酚型木素模型物具有较高的氧化还原电位,豆壳过氧化物酶(soybeanhullperoxidase,SHP,EC.1.11.1.7)通过依赖于过氧化氢的正常过氧化物酶催化循环不能氧化藜芦醇,但在还原型谷胱甘肽、Mn2+和有机酸络合剂存在下却可以通过不依赖于过氧化氢的氧化酶反应途径完成对藜芦醇的氧化,产物为藜芦醛,反应最适pH为4.2。动力学研究表明该反应遵循顺规序列反应机制;对藜芦醇的表观KM值为4.3mmol/L,对谷胱甘肽的表观KM值为4.8mmol/L。巯基还原剂二硫苏糖醇、L-半胱氨酸和β-巯基乙醇亦可替代还原型谷胱甘肽促进藜芦醇氧化     

人谷胱甘肽硫转化酶基因在链霉菌的高效表达

谷胱甘肽硫转化酶(GST)是一个具有广泛底物专一性,与一些化学致癌物代谢有关的酶的一个家族.它既有谷胱甘肽硫转化酶的活性,也有过氧化物酶的活性,能使谷胱甘肽分子上的巯基(SH)与广泛的亚硝基本类化合物结合,使这类致癌物转化成无毒的化合物.它亦能与细胞内源的一些阴离子化合物(如胆红素和亚铁血红素)结合并参与运输.在人体中,GST的缺乏常与新生儿非溶血性的胆红素积累而导致的病因有关,同时GST在保护生物体过氧化反应的损伤中也起着重要的作用.     

Glutathione and glutathione metabolizing enzymes in yeasts

Total glutathione content, glutathione peroxidase, glutathione transferase and glutathione reductase activities have been measured in 12 species of yeasts. All the strains tested contained glutathione, though in different amounts, as well as the above mentioned enzymes. To discriminate between the selenium-dependent and the selenium-independent form, glutathione peroxidase activity has been measured with both H₂O₂ and cumene hydroperoxide. Rhodotorula glutinis appeared to be the only strain in which the selenium-dependent form was not found, but this yeast exhibited the highest level of selenium-independent glutathione peroxidase activity as compared to the other strains.     

Development of Mechanisms of Protection Against Oxidative Stress in Doxorubicin-Resistant Rat Tumoral Cells in Culture

We have compared some mechanisms involved in the defense against doxorubicin-induced free radical damage in rat hepatoma and glioblastoma cell lines and their doxorubicin-resistant variants presenting an overexpression of the multidrug resistance gene.

Immediate in vivo production of malondialdehyde was minor and was not different in sensitive and resistant cells. Alpha-tocopherol was undetectable in all cell lines. Glutathione levels were not different in sensitive and resistant cells and these levels did not vary upon doxorubicin treatment. Resistant cells exhibited either a 50% decrease (hepatoma) or a 25% increase (glioblastoma) of glutathione-S-transferase activity. Glutathione reductase presented no important change upon acquisition of resistance. In contrast, selenium-dependent glutathione peroxidase activity was consistently 2-6-fold increased in the resistant cells, which suggests a magnification of protection mechanisms against hydroxyle radical formation from H₂O₂ in resistant cells. Depletion of glutathione levels by buthionine sulfoximine sensitized hepatoma resistant cells to doxorubicin, but had no effect on doxorubicin cytotoxicity to glioblastoma cells.     

Development of Mechanisms of Protection Against Oxidative Stress in Doxorubicin-Resistant Rat Tumoral Cells in Culture

We have compared some mechanisms involved in the defense against doxorubicin-induced free radical damage in rat hepatoma and glioblastoma cell lines and their doxorubicin-resistant variants presenting an overexpression of the multidrug resistance gene.

Immediate in vivo production of malondialdehyde was minor and was not different in sensitive and resistant cells. Alpha-tocopherol was undetectable in all cell lines. Glutathione levels were not different in sensitive and resistant cells and these levels did not vary upon doxorubicin treatment. Resistant cells exhibited either a 50% decrease (hepatoma) or a 25% increase (glioblastoma) of glutathione-S-transferase activity. Glutathione reductase presented no important change upon acquisition of resistance. In contrast, selenium-dependent glutathione peroxidase activity was consistently 2-6-fold increased in the resistant cells, which suggests a magnification of protection mechanisms against hydroxyle radical formation from H₂O₂ in resistant cells. Depletion of glutathione levels by buthionine sulfoximine sensitized hepatoma resistant cells to doxorubicin, but had no effect on doxorubicin cytotoxicity to glioblastoma cells.     

Elevation of glutathione levels and glutathione S-transferase activity in arsenic-resistant chinese hamster ovary cells

Summary Arsenic-resistant Chinese hamster ovary (CHO) cells were established by progressively increasing the concentration of sodium arsenite in culture medium. One of the resistant clones, SA7, was also cross-resistant to As(V), Zn, Fe(II), Co, and Hg. The susceptibilities to sodium arsenite in parental CHO cells, revertant SA7N cells, and resistant SA7 cells were correlated with their intracellular glutathione (GSH) levels and glutathione S-transferase (GST) activity. The resistance in SA7 cells was diminished by depletion of GSH in cells after treatment with buthionine sulfoximine. Furthermore, after reexposure of revertant SA7N cells to sodium arsenite, the intracellular GSH levels, GST activity, and resistance to sodium arsenite were raised to the same levels as SA7 cells. These data indicate that the elevation of intracellular GSH levels and GST activity in SA7 cells may be responsible for the resistance to arsenite. A p25 protein, which could be a monomer subunit of GST, accumulated in SA7 cells. In addition, an outward transport inhibitor, verapamil, indiscriminately increased the arsenite toxicity in resistant and parental cells. This work was supported in part by grant NSC77-0201-B001-31 from the National Science Council, Republic of China.     

Effect of Isoproterenol Administration on Rat Heart Glutathione Status

The intraperitoneal administration of 3, 10 and 80 mg/Kg isoproterenol produced in the cardiac muscle a dose dependent increase of GSH content and a slight elevation of GSSG content. In addition, the treatment with the catecholamine at the doses of 3 and 10 mg/Kg produced a slight decrease of the mixed glutathione disulfides level, whilst at the dose of 80 mg/Kg, this effect was more pronounced. These changes were not accompanied by modifications of the activities of the enzymes glutathione peroxidase, glutathione reductase and glutathione S-transferase.     

The effects of acute ethanol exposure and ageing on rat brain glutathione metabolism

Abstract

Binge alcohol consumption in adolescents is increasing, and it has been proposed that immature brain deals poorly with oxidative stress. The aim of our work was to study the effect of an acute dose of ethanol on glutathione (GSH) metabolism in frontal cortex, hippocampus and striatum of juvenile and adult rats. We have observed no change in levels of glutathione produced by acute alcohol in the three brain areas studied of juvenile and adult rats. Only in the frontal cortex the ratio of GSH/GSSG was increased in the ethanol-treated adult rats. GSH levels in the hippocampus and striatum were significantly higher in adult animals compared to young ones. Higher glutathione peroxidase (GPx) activity in adult rats was observed in frontal cortex and in striatum. Our data show an increased GSH concentration and GPx activity in different cerebral regions of the adult rat, compared to the young ones, suggesting that age-related variations of total antioxidant defences in brain may predispose young brain structures to ethanol-induced, oxidative stress-mediated tissue damage.     

Induction of glutathione S-transferase in the castor semilooper,Achaea janata (lepidoptera,Noctuidae) following fenitrothion treatment

Glutathione S-transferase activity was determined in the lepidopteran insect species,Achaea janata, during larval, pupal and adult stages following treatment with sublethal and lethal doses of fenitrothion. Both doses of insecticide produced significant induction of enzyme activity. The rate of induction of enzyme activity was not significantly different in insects that received sublethal and lethal doses of insecticide. Enzyme activity in the different stages of insecticide-treated insects was in the order pupa > adult > larva. However, the inducing effect of the insecticide was higher in larvae, than in pupae and adult. In the absence of induction, the level of enzyme was as much as 3 times higher in midgut tissue than in carcass. In larvae treated with sodium barbitone along with fenitrothion, the knock-down effect of the insecticide was delayed. This was attributed to the increased induction of glutathione S-transferase in the larvae treated with sodium barbitone. The level of reduced glutathione, a rate-limiting factor in the induction of glutathione S-transferase, changed in a cyclic manner in insecticide-treated larvae.     

Glutathione-related enzymes,glutathione and multidrug resistance

This review examines the hypothesis that glutathione and its associated enzymes contribute to the overall drug-resistance seen in multidrug resistant cell lines. Reports of 34 cell lines independently selected for resistance to MDR drugs are compared for evidence of consistent changes in activity of glutathione-related enzymes as well as for changes in glutathione content. The role of glutathione S-transferases in MDR is further analyzed by comparing changes in sensitivity to MDR drugs in cell lines selected for resistance to non-MDR drugs that have resulting increases in glutathione S-transferase activity. In addition, results of studies in which genes for glutathione S-transferase isozymes were transfected into drug-sensitive cells are reviewed. The role of the glutathione redox cycle is examined by comparing changes in elements of this cycle in MDR cell lines as well as by analyzing reports of the effects of glutathione depletion on MDR drug sensitivity. Overall, there is no consistent or compelling evidence that glutathione and its associated enzymes augment resistance in multidrug resistant cell lines.