白首乌对超氧阴离子自由基清除作用的研究

为深入探讨耳叶牛皮消块根（白首乌）的药理作用,指导其保健食品的研究与开发,采用化学发光法测定了去皮白首乌及白首乌栓皮对超氧阴离子自由基（Ｏ－２）的清除作用。结果表明,两者均能直接清除Ｏ－２,其ＩＣ５０分别为６９６９μｇ／ｍｌ和２２９８μｇ／ｍｌ,而且后者对Ｏ－２的清除能力比前者高出３０多倍,并同时测定了赤何首乌、西洋参、灵芝、当归、黄芪、菊花和竹叶。结果表明,去皮白首乌清除Ｏ－２的能力显著低于赤何首乌和灵芝,与西洋参和当归相差不大,显著高于黄芪、菊花和竹叶。而白首乌栓皮清除Ｏ－２的能力却高于其中任何一种中药。     

超氧阴离子自由基与嘧啶反应产物的量子化学研究

超氧阴离子自由基与嘧啶反应产物的量子化学研究班福强,戴柏青（哈尔滨师范大学,１５００８０）关键词超氧阴离子自由基;嘧啶;ＵＨＦ从头算超氧阴离子自由基（简称超氧自由基,）是一种重要的活性氧,与其他活性氧［如羟自由基（·ＯＨ）和过氧化氢（Ｈ２Ｏ２）等］一...     

余甘果汁清除超氧阴离子自由基的效能及人体试验初步观察

余甘子(Phyllanthus emblica L.)系大戟科叶下珠属落叶乔木或灌木。主要分布于南亚热带的印度、巴西、斯里兰卡、泰国和我国南方各省(区)。初食其     

针叶树越冬针叶光氧化与叶绿体超氧阴离子自由基

以Tiron为自旋探针,检出了越冬针叶叶绿体指示超氧阴离子自由基产生的TH~·ESR信号。此信号可由紫外辐射,也可由光合有效辐射产生。以单位叶绿素量为基数计算的信号强度比已报道的其它植物材料大2～3个数量级。红松叶绿体信号又大于其它针叶树。此信号可被外源SOD,抗坏血酸、乙醇和N_2抑制;随光照增加而增强,12月到次年5月期间又随时间而增强。上述结果表明红松针叶的冬季光氧化伤害与叶绿体中过量超氧阴离子自由基的产生有关。     

超氧阴离子自由基对绿豆黄化幼苗ACC合酶的影响

以0．5、5和50mmol／L的连二亚硫酸钠(Na2S2O4)为外源超氧阴离子自由基(O2^-)源,处理20min能明显提高绿豆黄化幼苗ACC合酶(ACC synthase,ACS,EC4．4．1．14)的活性。超氧阴离子自由基的特异性清除剂超氧化物歧化酶(superoxide dismutase,SOD)和1,4-二氮杂二环(2,2,2)辛烷[1,4-diazabicyclo(2,2,2)Octane,DABCO]能抑制Na2S2O4的这种作用,显示Na2S2O4产生的O2^-;引起了ACC合酶活性的升高。但Na2S2O4处理较长时间,ACC合酶活性开始下降,处理60min的ACC合酶活性明显低于对照,SOD和DABCO的加入有助于抑制ACC合酶活性的降低,表明超氧阴离子自由基对ACC合酶活性具有促进和抑制作用并存的“双重性”影响。研究表明,外源Na2S2O4处理20min能明显降低以S-腺苷蛋氨酸(S-adenosylmethionine,SAM)为底物的ACC合酶的Km值,处理60min反而提高了ACC合酶的Km值,表明O2^-;是通过改变ACC合酶对底物SAM的亲和力,从而影响其活性的。     

儿茶素对超氧阴离子自由基的清除及其自氧化作用研究

采用NBT光还原法证实了儿茶素具有清除O—·2 的作用 ,确定了儿茶素自氧化作用的最佳测定波长 ,并探讨了儿茶素溶液浓度和 pH对其自氧化作用的影响。     

光系统Ⅱ颗粒内单线态氧诱导产生超氧阴离子自由基

活性氧是生物体有氧代谢的必然产物。当机体处于疾病或胁迫条件下 ,活性氧的生成将更加活跃[1] 。由于类囊体膜是植物体光合放氧的器官 ,处于高浓度的氧环境中 ,同时类囊体膜上还存在着活跃的电子传递体系 ,因此类囊体膜是超氧阴离子自由基生成的活跃部位[2 ] 。最近的研究表明 ,ＰＳⅠ和ＰＳⅡ都是超氧阴离子自由基的生成部位[3] 。其中 ,对ＰＳⅠ生成超氧的机制研究已比较深入[2 .4 ] ,而对ＰＳⅡ生成超氧阴离子自由基的机制研究是在近几年内刚刚开展[3 ,5- 7] 。ＰＳⅡ中生成的另一活性氧是单线态氧。通常认为由于强光照射下ＰＳⅡ生成的…     

芥子碱硫氰酸盐清除超氧阴离子自由基作用的研究

目的:研究芥子碱硫氰酸盐清除超氧阴离子自由基的作用。方法:用化学发光法测定芥子碱硫氰酸盐清除超氧阴离子自由基的能力,并以抗坏血酸的清除能力做为对照,以IC50值(清除率为50%时的浓度值)作为评价指标。结果:芥子碱硫氰酸盐和抗坏血酸的IC50值分别为0.135 mmol/L和18.74 mmol/L,后者的IC50值约为前者的140倍。结论:芥子碱硫氰酸盐具有良好的抗氧化作用,可以作为天然抗氧化剂进行开发。     

植物中超氧阴离子自由基测定方法的改进

通过对植物超氧阴离子自由基测定反应中动力学曲线的分析, 确定了最佳的反应介质、反应参数和羟胺浓度, 以三氯甲烷代替乙醚作为植物色素萃取试剂, 克服了植物超氧阴离子测定中存在的诸多问题, 提高了测定结果的准确性、重复性和可比性。     

Free Radicals and the Ischemia-Evoked Extracellular Accumulation of Amino Acids in Rat Cerebral Cortex

The effects of free radical generating systems on basal and ischemia/reperfusion-evoked release of amino acids into cortical superfusates was examined in the rat using the cortical cup technique. Xanthine oxidase plus xanthine significantly enhanced GABA levels 358 fold over controls during 20 min of four vessel occlusion. Glutamate and phosphoethanolamine release following reperfusion were also elevated. Prostaglandin synthase plus arachidonic acid significantly enhanced the ischemia-evoked release of all amino acids (aspartate 360 fold; glutamate 433 fold; glycine 6 fold; GABA 689 fold; phosphoethanolamine 69 fold) and increased the pre-ischemic levels of glutamate, glycine and phosphoethanolamine. Administration of H₂O₂ plus ferrous sulfate significantly elevated both pre-ischemic amino acid release and ischemia-evoked release. A role for free radical generating systems in the development of ischemic injury is supported by the ability of superoxide dismutase plus catalase to reduce ischemia-evoked amino acid efflux into cortical superfusates. Thus, the species of free radical produced, as well as the amount generated, may alter the pattern of amino acid release under both ischemic and non-ischemic conditions.     

Age-Dependent Resistance to the Toxic Effects of Paraquat in Relation to Superoxide Dismutase Activity in Rat Lung

Young rats are much more resistant to the toxic effects of orally-administered paraquat than are older rats. The possible relation of this phenomenon to the activity of superoxide dismutase in lung is discussed.     

几种外源因子对大豆幼苗SOD活性的影响

高氧能促使大豆幼苗细胞内产生O_2~+速率增加,同时又使幼苗内SOD活性水平提高,以减轻O_2~+增加所引起的细胞伤害。高氧诱发O_2~+同时发生在叶绿体、线粒体和细胞溶质中,与细胞呼吸水平无明显相关。棓酸丙酯有清除体内O_2~+的能力,当浓度在1μmol/L时,能减轻大豆幼苗的氧伤害。相反,DDC是SOD的有效抑制剂,当它的浓度大于5 mmol/L时,显著抑制大豆幼苗SOD活性,增加了体内O_2~+的积累,影响了幼苗的正常生长以及幼苗氧伤害的加剧。     

Induction of Manganese Superoxide Dismutase by Cytokines and Lipopolysaccharide in Cultured Mouse Astrocytes

Abstract: To determine whether cytokines or lipopolysaccharide (LPS) are involved in the induction of superoxide dismutase (SOD) in the nervous system, we examined the effects of these substances on the levels of SOD in cultured mouse astrocytes. Treatment of astrocytes with 10² to 10⁴ U/ml tumor necrosis factor-α for 3 days increased the levels of Mn SOD in a dose- and time-dependent manner to as much as six times the level under nontreated conditions. Treatment with 1.0 µg/ml LPS for 3 days elicited a fourfold increase in levels of Mn SOD, and the effect of LPS was also dose dependent. Furthermore, Mn SOD in astrocytes was induced by a 3-day exposure to interleukin-1α at concentrations of 10² or 10³ U/ml. However, these stimuli had no effect on levels of copper-zinc SOD (Cu/Zn SOD) in astrocytes. By contrast, interferon-γ did not change the levels of either Mn or Cu/Zn SOD in the cells. The results indicate that the selective induction of Mn SOD by cytokines and LPS, which has been observed in nonnervous tissues, may also occur in nervous tissues. The induction of Mn SOD may represent a mechanism for protection of cells from oxidative stress.     

In vitro Damage to Rat Lens by Lumazine and Xanthine Oxidase: Prevention by Superoxide Dismutase

Intact rat lenses incubated with lumazine and xanthine oxidase are physiologically damaged as evidenced by a decrease in the net accumulation of rubidium ions against a concentration gradient. Superoxide dismutase protected the tissue against this damage. These experiments, therefore, demonstrate the susceptibility of the lens tissue to O₂^?_? injury under ambient and nonphotochemical conditions, suggesting a possible implication of this radical in the tissue in vivo and eventual cataract formation. The lumazine/xanthine oxidase system which is known to cause oxygen reduction predominantly by the monovalent route, producing superoxide, appears quite suitable to evaluate the toxicity of O₂^?_? to the tissues in vitro.     

From Haemocuprein to Copper-Zinc Superoxide Dismutase: A History on the Fiftieth Anniversary of the Discovery of Haemocuprein and the Twentieth Anniversary of the Discovery of Superoxide Dismutase

Haemocuprein was discovered fifty years ago by T. Mann and D. Keilin as a copper protein of red blood cells, later named erythrocuprein. Superoxide dismutase was discovered twenty years ago by J.M. McCord and I. Fridovich as an enzymatic activity in preparations of carbonic anhydrase or myoglobin that inhibited the aerobic reduction of cytochrome c by xanthine oxidase. Astonishingly the superoxide dismutase proved to be haemocuprein. Around this time zinc was found in haemocuprein, in equimolar amount to the copper. Haemocuprein thus became copper-zinc superoxide dismutase after thirty years as an obscure cupropro-tein of red blood cells. This historical article is a tribute to the achievement of J.M. McCord and I. Fridovich. Their discovery of superoxide dismutase revolutionized the study of oxygen free-radicals in biochemistry.     

Superoxide Production by the Mitochondrial Respiratory Chain

This mini-review describes the role of different mitochondrial components in the formation of reactive oxygen species under normal and pathological conditions and the effect of inhibitors and uncouplers on superoxide formation.     

磷脂化修饰影响超氧化物歧化酶细胞亲和力的研究

目的:研究磷脂化修饰对重组人超氧化物歧化酶(SOD)进入人冠状动脉内皮细胞(HCAEC)、人心肌细胞(HCM)能力的影响。方法:分别运用流式细胞术和蛋白印迹分析磷脂化修饰的超氧化物歧化酶(PC-SOD)和SOD与HCAEC、HCM的结合能力,并用激光共聚焦显微术分析修饰前后的SOD可显著增强PC-SOD与细胞的亲和力,并可显著增强PC-SOD进入人冠状动脉内皮细胞和人心肌细胞的能力。     

新生SD大鼠皮质神经元的体外培养和鉴定

目的：建立高纯度的新生SD大鼠皮质神经元原代培养方法。方法：取24h内的新生SD大鼠皮质,用木瓜酶和DNaseⅠ共同消化,5％胎牛血清终止消化,吹打分离组织获得单细胞悬液,进行细胞计数,用无血清DMEM／F12种植培养,4h后换成用无血清Neurobasal配制的维持培养液继续培养,尼氏小体染色和免疫荧光法鉴定神经元的纯度。结果：培养第10d,神经元胞体饱满,结构清晰完整,光晕明显,折光性强,可见粗长的树突和轴突,相邻细胞形成紧密网状联系,神经元纯度达到96％以上。结论：经改良和优化,无须添加阿糖胞苷抑制胶质细胞的生长即能够获得生长状态良好、高纯度的神经元。     

Cyclooxygenase-Mediated Generation of Free Radicals During Hypoxia in the Cerebral Cortex of Newborn Piglets

Previous studies have demonstrated that free radicals are formed under hypoxic conditions in newborn piglet brain. To test the hypothesis that the cyclooxygenase pathway serves as a source of free radical generation during hypoxia studies were performed on 24 piglets divided into four groups. Six saline (group 3) and six indomethacin treated (group 4) were exposed to hypoxia (FiO2 0.05-0.07) for 60 min. Cerebral hypoxia was documented biochemically by determination of ATP and phosphocreatine. Fluorescent compounds and conjugated dienes were determined as indices of lipid peroxidation. Free radical formation was determined by using n-tert butyl phenyl nitrone (PBN) as a spin trap agent and measuring spin adduct formation in duplicate using a Varian E-109 spectrometer. Groups 1 and 2 (normoxic) showed no spin adduct formation. Group 3 showed a significant increase in spin adduct formation compared to normoxia (372+/-125 vs. 63+/-15, P<0.001). Hypoxic animals pretreated with indomethacin had a spin adduct level of 197+/-132 and were similar to normoxic animals. ATP/PCr levels were the same in groups 3 and 4 denoting the same degree of cerebral hypoxia in all hypoxic animals. Conjugated dienes increased significantly during hypoxia as compared to normoxia (0.142+/-0.017 vs. 0.0+/-0.0) and were decreased insignificantly with indomethacin treatment. Fluorescent compounds were not significantly different among the four groups. Na+,K+-ATPase activity decreased during hypoxia but was not preserved in hypoxic animals pretreated with indomethacin. These data provide direct evidence of the presence of free radicals during hypoxia and the contribution of cyclooxygenase metabolism to their formation.