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Dolmatova L. S. Eliseykina M. G. Timchenko N. F. Kovaleva A. L. Shitkova O. A. 《中国海洋湖沼学报》2003,21(4):293-304
Pure fraction (92%–95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38%)-FPMC- of the holothurianEupentacta fraudatrix' (Holothuroidea, Dendrochirota) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified
by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin ofYersinia pseudotuberculosis (TST) at different concentrations (0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation.
In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction
after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively.
Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared
to that under toxin treatment alone. TST stimulated superoxide dismutase activity in concentration-dependent manner (maximum
at 0.5 μg/ml concentration in FP) after 24 h treatment, and this stimulation was prevented by a commercial catalase. Plant
lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin
stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously,
ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in
holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production
by these cells may be one of the mechanisms of antibacterial protection of holothurians.
This work was financially supported by the Russian Foundation for Fundamental Research Grant (No. 00-04-48949). 相似文献
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Florence Geret Angela Serafim Luisa Barreira Maria Joo Bebianno 《Marine environmental research》2002,54(3-5)
Copper (Cu) is an essential element for biological systems, however, when present in excess, is toxic. Metallothioneins can play an important role in Cu homeostasis and detoxification. Moreover, Cu can catalyse the production of toxic hydroxyl radicals that cause lipid peroxidation but defence systems in the cells can limit the oxidative damage. The present study was performed to investigate the effect of three Cu concentrations (0.5, 2.5 and 25 μg l−1) on the response of antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathion peroxidase and total glutathion peroxidase), total proteins, metallothioneins (MT), malondialdehyde (MDA) concentrations in the gills of the clam, Ruditapes decussatus. The activity of antioxidant enzymes and total proteins, MT and MDA concentrations were measured in the gills of the clams after 1, 3, 7, 14, 21 and 28 days of Cu exposure. Results indicate that Cu only induces an imbalance in the oxygen metabolism during the first week of Cu exposure due to a decrease in mitochondrial SOD and CAT, selenium-dependent and total glutathion peroxidase activities. Cu also causes lipid peroxidation, measured by the MDA formation, that was Cu dependent. In the gills of clams exposed to 25 μg Cu/l, the excess of Cu triggers the induction of MT synthesis after 3 days of exposure. 相似文献
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检测了受三丁基锡(tributyltin,TBT)污染的牡蛎的吞噬细胞活力、超氧化物歧化酶活性、过氧化氢酶活性、脂质过氧化作用等生理生化指标。实验结果表明,牡蛎血细胞的吞噬活力随着TBT浓度的增加而下降;对不同TBT浓度下牡蛎的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性测定发现,在较低浓度下(TBT质量浓度<10μg/L),SOD和CAT的酶活力被抑制,随着TBT浓度的增大,SOD和CAT的比活力也随着增大,但当TBT达到较高浓度时,SOD和CAT的比活力开始下降,而过氧化脂质(LPO)却随着TBT的浓度的增加而增大。 相似文献
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极大螺旋藻铁型超氧物歧化酶的纯化及性质 总被引：3，自引：0，他引：3
极大螺旋藻 Ｓｐｉｒｕｌｉｎａｍａｘｉｍａ藻体ＳＯＤ聚丙烯酰胺梯度凝胶电泳呈现４条同工 酶带, Ｈ_ ２Ｏ_２明显抑制该酶活性, ＫＣＮ对酶活性无影响,确认为 Ｆｅ－ＳＯＤ经硫酸铵分部盐析、离子交 换柱层析及凝胶过滤,纯化到电泳单斑点均－程度．纯化的Ｆｅ－ＳＯＤ分子量为３９．３ＫＤ,亚基分子 量为２０ＫＤ金属元素分析表明,每个亚基含０ ５５个Ｆｅ原子,该酶在紫外区最大吸收峰值为 ２７５．８nｍ、该酶氨基酸组成与蓝、绿藻和高等植物的Ｆｅ－ＳＯＤ相似,但它含有较高的丙氨酸,酸性氨 基酸和碱性氨基酸比值与低等植物及原核生物相近而明显高于高等植物． 相似文献
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免疫多糖对栉孔扇贝酸性磷酸酶、碱性磷酸酶和超氧化物歧化酶活性的影响 总被引：24，自引：1，他引：23
在栉孔扇贝的血清、血细胞和肝脏提取液中均发现有酸性磷酸酶（ＡＣＰ）、碱性磷酸酶（ＡＬＰ）和超氧化物歧化酶（ＳＯＤ）活性,其中,ＡＣＰ和ＡＬＰ在肝脏中的活性最高。采用１．０％的虫草多糖或海藻多糖作为免疫药物,对栉孔扇贝进行注射刺激后,发现血清中的ＡＣＰ、ＡＬＰ和ＳＯＤ活性有显著提高,血细胞中的ＡＣＰ和ＳＯＤ活性也有所增加,而肝脏提取液的这三种酶活性的变化不很明显。研究认为这两种多糖具有增强栉孔扇贝免疫活性的作用,在水产养殖业中有一定的开发价值。 相似文献
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嗜热栖热菌(Thermus thermophilus)wl的超氧化物歧化酶基因在Escherichia coli BL21(DE3)中进行表达,采用Ni-NTA agarose亲和层析获得重组超氧化物歧化酶,比活力达565 U/mg.重组酶的相对分子质量为110.9 kDa,亚基相对分子质量为28.3 kDa,该酶为同聚四聚体蛋白.重组酶对氯仿-乙醇和SDS敏感,对H2O2不敏感,并且它的紫外最大吸收波长位于279 nm,判断其属于锰超氧化物歧化酶类型.1.0 mmol/dm3Mn2+对重组酶有激活作用,同浓度的其他金属离子Mg2+、Ca2+、Ba2+、Co2+、Cu2+、Zn2+和Fe2+对重组酶活力都有抑制作用,其中以Co2+的作用最强,抑制了约64%的酶活性.重组酶在70℃下处理1 h,保持原有活力的60%,即使在90℃下处理1 h,仍保持原有活力的35%.在pH值为4.0～11.0的缓冲液中25℃下处理1 h,保持65%以上的原始活力.由此可见,重组锰超氧化物歧化酶具有高的热稳定性和宽pH稳定性,在工业上具有巨大的应用潜力. 相似文献
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近海温泉中嗜热菌Geobacillus sp. ZH1锰超氧化物歧化酶的克隆与表达 总被引：1，自引：0，他引：1
将嗜热菌Geobacillus sp. ZH1的超氧化物歧化酶(supseroxide dismutase,SOD)基因插入表达载体pET-32α(+),在Escherichia coli BL21(DE3)中进行表达,并利用亲和层析纯化重组超氧化物歧化酶.将制备的脱辅SOD进行Mn2+和Fe2+金属重构后,得到的Mn2+重构SOD的比活力达668U/mg,Fe2+重构Fe-SOD没有活性,说明ZH1菌株的超氧化物歧化酶为Mn-SOD.凝胶过滤及SDS-PAGE分析显示,Mn2+重构SOD为同聚二聚体,亚基分子量为71.7 kDa.这些研究结果为进一步研究该酶的生化特性及酶的定向进化研究奠定了良好的基础. 相似文献