Pedrajas J R, Peinado J, López-Barea J
Departamento de Bioquímica y Biología Molecular, Instituto de Biología Básica y Aplicada, Facultad de Veterinaria, Universidad de Córdoba, Spain.
Chem Biol Interact. 1995 Dec 22;98(3):267-82. doi: 10.1016/0009-2797(95)03651-2.
Fish (Sparus aurata) were intraperitoneally injected with model xenobiotics and several biomarkers of oxidative stress were analysed after 2 and 7 days exposure. The levels of soluble thiobarbituric acid reactive substances (TBARS) increased markedly in animals treated with polar xenobiotics, CuCl2 or paraquat; exposure to the apolar xenobiotics, dieldrin or malathion, enhanced significantly the microsomal TBARS while decreasing the microsomal glutathione transferase activity. The specific superoxide dismutase (SOD) activity increased in Cu(II)-injected animals but diminished in fish exposed to paraquat. After isoelectrofocusing separation and activity staining cell-free extracts of fish exposed to Cu(II), dieldrin or malathion displayed two new Cu,Zn-SOD isoforms of intermediate pI. An additional Mn-SOD was observed in dieldrin-injected fish, but only a faint new acidic isoform was observed in paraquat-injected animals. The new SOD bands were reproduced in vitro by incubation of cell-free extracts with systems generating superoxide anion or hydrogen peroxide and with a tert-butyl hydroperoxide/ADP-Fe system. Metallothionein induction was observed in Cu(II) or paraquat-exposed fish, but not in animals injected with apolar xenobiotics. So, the new SOD bands are possibly oxidized forms of this enzyme and can be considered as useful early biomarkers of oxidative stress due to transition metals or organic xenobiotics.
将模型异生物质腹腔注射到金头鲷体内，并在暴露2天和7天后分析几种氧化应激生物标志物。用极性异生物质、氯化铜或百草枯处理的动物中，可溶性硫代巴比妥酸反应性物质（TBARS）水平显著升高；暴露于非极性异生物质狄氏剂或马拉硫磷中，微粒体TBARS显著增加，而微粒体谷胱甘肽转移酶活性降低。注射铜（II）的动物中特异性超氧化物歧化酶（SOD）活性增加，但暴露于百草枯的鱼中该活性降低。在等电聚焦分离和活性染色后，暴露于铜（II）、狄氏剂或马拉硫磷的鱼的无细胞提取物显示出两种新的中等pI的铜锌超氧化物歧化酶同工型。在注射狄氏剂的鱼中观察到一种额外的锰超氧化物歧化酶，但在注射百草枯的动物中仅观察到一条微弱的新酸性同工型。通过将无细胞提取物与产生超氧阴离子或过氧化氢的系统以及叔丁基过氧化氢/ADP-铁系统孵育，可在体外重现新的超氧化物歧化酶条带。在暴露于铜（II）或百草枯的鱼中观察到金属硫蛋白的诱导，但在注射非极性异生物质的动物中未观察到。因此，新的超氧化物歧化酶条带可能是该酶的氧化形式，可被视为由过渡金属或有机异生物质引起的氧化应激的有用早期生物标志物。
