三种鼠科动物血浆超氧化物歧化酶和乳酸脱氢酶以及血浆蛋白成分的比较分析

本文利用SOD和LDH在一块凝胶板上同时显色的优点,电泳分析黑线姬鼠(Apodemus agrarius)、褐家鼠(Rattus norvegicus),黄胸鼠(Rattus flavipectus)血浆SOD和LDH同工酶以及血浆蛋白成分。结果表明,LDH为有色区带,SOD为无色透明区带,而且SOD比LDH泳动快,两类区带互不重迭,以致3种鼠血浆中SOD和LDH的图谱清晰可辨。由于黑线姬鼠的SOD和LDH的泳动度不同于其余2种鼠,周此3种鼠的图谱各具特征。3种鼠血浆蛋白中的前清蛋白和铁传递蛋白区带数和泳动度亦具有明显差异,说明上述酶分子亚基和血浆蛋白分子的化学结构不同,成为种间差异在分子水平上典型的表现。     

LDH和SOD同工酶同时显色法及其运用实例

介绍了一种新颖的同工酶染色法,使得LDH和SOD同工酶在同一块凝胶板上同时显色,既经济又快速,增加了同工酶分析的精确性,并且介绍了本方法在实际研究中的运用。     

乳酸脱氢酶(LDH）同工酶在小白鼠胚胎发育过程中的变化

同工酶是一组能催化同一种生化反应但分 子组成不同的酶。乳酸脱氢酶(lactate dehydrogenase, 简称LDH）是参与糖酵解的酶。Market 等曾提纯组织LDH,并进行淀粉凝胶电泳 的分析,确定LDH有5类同工酶,从阳极到 阴极显示出LDH LDH LDH3, LDH,及 LDH,的5条区带,并指出LDH 同工酶电泳 图在不同组织和不同的发育阶段是不同的。     

羚牛、羊、牛血清同工酶的比较研究

本文报道了羚牛、同羊及秦川牛的血清乳酸脱氢酶(LDH)、酯酶(Es)聚丙烯酰胺凝胶的不连续电泳图谱,并对这3种动物的酶谱进行了分析比较,从酶谱的区带数目、泳动率、相对含量及染色强度来看,均表明3者各具有特征电泳图谱,且羚牛与同羊图谱的相似程度较秦川牛接近。     

超氧化物歧化酶同工酶分析法用于兽类分类的试探

本文以蝙蝠的5种组织和鼠类的红细胞为材料,从方法上探讨SOD同工酶的分析比较对于兽类分类的价值和意义。实验结果表明SOD同工酶的泳动速度和区带的分布组成,种与种之间有明显的区别,作为一个辅助的工具酶对于兽类的演化和物种的鉴定比较适用。此外,同种的个体间以及同个体的组织间未发现明显区别,这预示在分类工作中不必强调被分析个体数的多寡,也不必进行各组织的全面分析,有利于取材不易的或不能杀死取材的珍稀兽类物种间的比较分析。     

乳酸脱氢酶与酯酶同工酶同板染色法

介绍一种在同一块凝胶板上染乳酸脱氢酶(LDH)与酯酶(EST)的染色方法. 该同板染色法利用两种同工酶显色反应互不干扰和颜色不同的特点, 先染LDH, 后染EST, 可以在同一块胶板上得到两种同工酶清晰的酶带, 每一种酶的酶带与单板染色的酶带完全一样. 这种染色法, 能节省同工酶分析所需的试剂、时间和经费, 也便于样品的鉴定与比较, 是一种经济有效的方法. 此方法, 同样适用于苹果酸脱氢酶(MDH)与酯酶等同工酶的同板染色.     

四种园蛛同工酶的比较研究

本文采用聚丙烯酰胺凝胶电泳的方法,对叶斑园蛛、角类肥蛛(黑、白2种体色)、大腹园蛛、黑斑园蛛4种园蛛科蜘蛛的酯酶(EST)同工酶、乳酸脱氢酶(LDH)同工酶、超氧化物歧化酶(SOD)同工酶进行了比较研究。结果表明,4种园蛛的EST、LDH具有明显的种族特异性,而SOD在种间基本没有差别。不同种蜘蛛都有各自的EST、LDH同工酶谱型,可以用它们作为识别物种的附加指标。     

大豆黄酮对衰老小鼠脑组织SOD、LDH同工酶的影响

利用SOD和LDH同工酶电泳分析,研究大豆黄酮对衰老小鼠的抗氧化作用。结果显示大豆黄酮没有改变SOD和LDH同工酶谱的特征,但对因衰老引起的小鼠脑组织LDH和SOD同工酶活性、各组分的相对活性和比活力的变化有不同程度的改善作用,即LDH同工酶中LDH-2、LDH-3的活性明显下降,LDH-1的活性下降最为明显,而LDH-4的活性有所下降,但不显著,LDH-5的活性几乎没有变化,SOD同工酶的SOD-1和SOD-2的活性有不同程度的升高。这表明大豆黄酮是通过抑制LDH同工酶H亚基的合成来降低LDH的活性,而对M亚基的合成没有影响,并且能够促进SOD同工酶SOD-1和SOD-2的合成,不影响其遗传稳定性。     

牛蛙乳酸脱氢酶同工酶的分离纯化及其性质研究

牛蛙心脏中乳酸脱氢酶在聚丙烯酰胺凝胶电泳上显示3种同工酶区带,分别命名为LDH1、LDH2、LDH3,其中LDH1的活力占绝对优势.采用HiTrap^TM Blue HP 亲和层析和DEAE-Sephadex A离子交换层析对牛蛙骨骼肌中的LDH3进行了分离纯化.纯化的LDH3比活力为295 U/mg,Km NADH=0.028,Km丙酮酸=1.242,在SDS-PAGE上显示两条带,提示该同工酶是由两种亚基组成的,亚基的分子量分别为35.3 kD和37.6 kD.     

六种鼠组织中过氧化物酶和酯酶变异的比较

本文研究姬鼠属（Ａｐｏｄｅｍｕｓ）２种鼠和鼠属（Ｒａｔｕｓ）４种鼠的心组织过氧化物酶和肝组织酯酶同工酶等位基因的变异。结果表明,６种鼠心组织过氧化物酶皆为４条区带,其中社鼠和白腹巨鼠的酶带泳动速度略快于褐家鼠和黄毛鼠的酶带;鼠属４种鼠的酶带间距大致相同,而姬鼠属的酶带间距略大于鼠属,这种谱型的区别就所分析的６种鼠而言,可以认为是该同工酶表现的属间区别。肝组织酯酶区带较多,其酶带数和泳动速度的差异具有明显的种间特征;泳动最快的酶带在姬鼠中是２条,在鼠属中是１条,可以认为是属间特征的区别。从９项生化指标的异同作动物配对比较,表明褐家鼠与黄毛鼠亲缘上较近,社鼠与白腹巨鼠亲缘上也较近,而且白腹巨鼠在进化上可能要比其余３种鼠属动物更接近于姬鼠属     

The immunohistochemical localization of superoxide dismutase activity in the avian epithelial growth plate

Summary Superoxide dismutase (SOD) is a scavenger enzyme which catalyses the dismutation (reduction—oxidation) of the superoxide anion (O₂ ^–), a toxic free radical generated during normal cellular respiration. Light microscopy employing immunohistochemistry was utilized for localizing SOD activity in the chick epiphyseal cartilage. Antibodies to mammalian liver CuZn—SOD were prepared and the avidin—biotin—peroxidase technique (ABC complex) was utilized to localize activity for this enzyme in the growth plate cartilage. The localization of enzyme activity varied in accordance with the characteristic zonation pattern of the growth plate (zone of proliferation, zone of maturation, zone of cell hypertrophy and zone of matrix calcification). In the upper regions of the epiphyseal cartilage (the zones of proliferation and maturation), where the vascularity is poor and the oxygen tension low, SOD activity was localized within the chondrocytes. No extracellular activity was observed. However, in the lower regions of the growth plate (the zones of cell hypertrophy and matrix calcification), where both the vascularity and the oxygen tensions are increased, SOD activity was intense in both the chondrocytes and the surrounding extracellular matrix. Thus, the distribution of SOD enzyme activity in this tissue seems to vary in accordance with the level of oxygen present. The significance of the extracellular SOD activity, seen in the lower aspects of the growth plate cartilage, may indicate the sensitivity of matrix components, especially collagen, to toxic free radicals such as the superoxide anion.     

Changes in superoxide dismutase activity in the presence of electron donors and acceptors

The activity of superoxide dismutase (SOD) from bovine erythrocytes was measured by the inhibition of nitrotetrazolium blue reduction rate in superoxide anion radical generation systems--xanthine/xanthine oxidase of NADH/phenazine methasulfate. The enzyme activity increases in the presence of compounds acting as electron donors in radical-involving reactions and decreased in the presence of compounds possessing the properties of electron acceptors. Activation of SOD by electron donors and its inhibition by electron acceptors was dependent on the concentration of the above compounds. In the absence of SOD electron donors and acceptors did not change the rate of tetrazolium blue reduction by superoxide anion radicals. The role of the new type of SOD regulation for the enzyme functioning in the cell is discussed.     

水母胶原蛋白抗氧化活性研究

提取水母胶原蛋白,观察其体内外抗氧化作用。体外采用邻苯三酚自氧化法观察水母胶原蛋白对超氧阴离子(·O2-)的清除作用,采用Fenton反应体系观察对羟自由基(·OH)的清除作用;体内观察水母胶原蛋白对小鼠血清、肝脏、大脑中超氧化物歧比酶(SOD)、过氧化氢酶(CAT),丙二醛(MDA)含量的影响。结果表明:水母胶原蛋白对·O2-、·OH具有良好的清除作用,呈一定的剂量依赖关系,能明显提高小鼠血清、肝脏、大脑中的SOD、CAT含量,能在一定程度上降低血清、肝脏、大脑中的MDA水平。因此,水母胶原蛋白具有显著的抗氧化生物活性。     

六种鼠乳酸脱氢酶和超氧化物歧化酶位点变异的研究

用电泳比较分析了鼠科动物６个种的组织ＬＤＨ同工酶的Ａ、Ｂ、Ｃ亚基和ＳＯＤ同工酶位点的种属差异。结果表明,ＬＤＨ的Ａ和Ｂ亚基在进化上的可变性程度不同。在被分析的２个属共６种鼠中,首次发现Ｂ亚基的结构具有属的特征,同属的鼠种Ｂ亚基泳动度相同,在垂直凝胶电泳板上处于同一泳动线上。不同属的鼠其Ｂ亚基泳动度不同,家鼠属的鼠Ｂ亚基比姬鼠属的泳动快;Ａ亚基的结构既有属的特性,更具有种的特征,同属不同种的鼠Ａ亚基泳动速度不同,表现为ＬＤＨ同工酶带的间距不同。ＬＤＨ的Ｃ亚基结构因鼠种不同而异,由Ｃ亚基组成的ＬＤＨ－Ｘ带黄毛鼠位于自身的ＬＤＨ－３和ＬＤＨ－４之间,社鼠和褐家鼠此带位于自身的ＬＤＨ－４和ＬＤＨ－５之间,白腹巨鼠此带位于自身的ＬＤＨ－５以外的阴极端,与黑线姬鼠此带的位置特征相同。ＳＯＤ同工酶带只表现种的差异,其３条带泳动速度的改变具有协同而变的共同特点。被分析的鼠种经配对法比较生化特征的相同程度,初步表明黄毛鼠亲缘上与褐家鼠比较近,白腹巨鼠亲缘上又比其余３种鼠更近于姬鼠属。     

两个雌核发育白鲢群体同工酶分析及遗传标记的确定

取源于武汉两个不同渔场两尾白鲢的卵子,经紫外照射遗传物质失活的鲤鱼精子刺激雌核发育和热休克诱导第二极体保留的基因组操作技术,获得了两个不同的人工雌核发育白鲢群体。采用聚丙烯酰胺垂直板电泳技术,分析了这两个不同人工雌核发育白鲢群体(分别称为Hy-G1和Hy-G2)内不同个体的肝脏、肌肉组织以及红细胞中乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、酯酶(EST)、超氧化物歧化酶(SOD)等几种同工酶的表达谱式,并与普通繁殖的同龄白鲢进行了比较。结果表明,各个雌核发育白鲢群体内不同个体间的酶谱表现出很大程度的一致性,具较高的纯合度,而两个不同雌核发育群体Hy-G1和Hy-G2之间表现有明显差异。特别是肝脏和肌肉组织迁移率较快的酯酶谱带等以其稳定的差异,建议作为区分这两个人工雌核发育白鲢群体的生化遗传标记。       

Endothelial Cells as A Source of Oxygen-Free Radicals An Esr Study

Endothelial cells were subjected to anoxia/reoxygenation in order to simulate some of the free radical mechanisms occurring in ischaemialreperfusion. With ESR and spin trapping using the spin traps 5.5-dimethyl-l-pyrroline-l-oxide (DMPO) and 3,3,5,5-dimethyl-l-pyrroline-l-oxide (M₄PO), the results show that upon reoxygenation of endothelial cells, following a period of anoxia, these cells generate superoxide (0₂). Cytotoxicity of the spin traps was measured by standard trypan blue exclusion methods. Cell injury or death was measured at various times during reoxygenation by lactate dehydrogenase (LDH) release. Experiments using oxypurinol, SOD, CAT and a combination of SOD and CAT show that while oxypurinol partially prevents spin adduct formation. the combination of SOD and CAT is more effective in doing so. These results suggest that the majority of the oxygen radicals produced by endothelial cells are done so exogenously. The results also suggest that endothelial cells are not only a source of oxygen radicals but also a target.     

Endothelial Cells as A Source of Oxygen-Free Radicals An Esr Study

Endothelial cells were subjected to anoxia/reoxygenation in order to simulate some of the free radical mechanisms occurring in ischaemialreperfusion. With ESR and spin trapping using the spin traps 5.5-dimethyl-l-pyrroline-l-oxide (DMPO) and 3,3,5,5-dimethyl-l-pyrroline-l-oxide (M₄PO), the results show that upon reoxygenation of endothelial cells, following a period of anoxia, these cells generate superoxide (0₂). Cytotoxicity of the spin traps was measured by standard trypan blue exclusion methods. Cell injury or death was measured at various times during reoxygenation by lactate dehydrogenase (LDH) release. Experiments using oxypurinol, SOD, CAT and a combination of SOD and CAT show that while oxypurinol partially prevents spin adduct formation. the combination of SOD and CAT is more effective in doing so. These results suggest that the majority of the oxygen radicals produced by endothelial cells are done so exogenously. The results also suggest that endothelial cells are not only a source of oxygen radicals but also a target.     

海藻中清除氧自由基的物质

新鲜海藻的提取液含有超氧物歧化酶（SOD）活性物质,能清除超氧自由基（O2-）。海藻的SOD活性通常为60—280Ug-1FW,而在孔石莼（Ulvapertusa）、江蓠（Gracilariaverrucosa）和凤尾菜（G．eucheumoices）中活性较高,约为300Ug-1FW。一般来说,海藻的SOD活性和稳定性为：绿藻＞红藻＞褐藻。绿藻的SOD以CuZn-型为主,而蓝藻的SOD以Fe-型为主。以江蓠琼枝（Eucheumagelatinae）提取液作PAGE并SOD活性染色时,除了观察到SOD同工酶带之外,还发现在前沿指示剂附近有一区域,此区域与高效自由基清除剂SPD（Superphycodismutas）的电泳行为和对氮蓝四唑（NBT）负染色的抑制相同,可能两者为同一种物质。     

Source of superoxide anion radical in aerobic mixtures consisting of NAD[P]H, 5-methylphenazinium methyl sulfate and nitroblue tetrazolium chloride

The source of superoxide anion radical (O2-.) in aerobic mixtures consisting of NAD[P]H, 5-methylphenazinium methyl sulfate (or its 1-methoxy derivative) and tetrazolium salt was investigated using superoxide dismutase (SOD), Mn(II), ferricytochrome-C, and epinephrine as probes. NAD[P]H + phenazine + O2 was found to reduce nitroblue tetrazolium, iodonitrotetrazolium, and thiazolyl blue in a manner sensitive to agents that dismutase O2-., viz., SOD and Mn(II). It also mediated the reduction of ferricytochrome-C, and augmented the autooxidation of epinephrine to the adrenochrome, without a tetrazolium salt present in the medium. The autooxidation of epinephrine, but not the reduction of ferricytochrome-C, was found to be sensitive to SOD. Nitroblue tetrazolium, either singly or in combination with SOD, did not stimulate the reduction of ferricytochrome-C. The oxidation of NADH, mediated by a catalytically low concentration of phenazine(+O2), was augmented two-fold by SOD. These observations are consistent with, and lend support to, a scheme of redox events (Scheme-3) wherein it is proposed that the source of O2-. in the NAD[P]H + phenazine + tetrazolium(+O2) system is the reduced phenazine, that the tetrazoinyl radical (a one-electron reduction product of tetrazolium) may not reduce O2 to O2-., that the redox reaction between semiquinone radicals of phenazine and O2 is reversible, and that the disproportionation of semiquinone radicals constitutes an important rate-limiting reaction in the expression of phenazine redox couple.