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宽吻凯门鳄血液中氧化损伤和抗氧化防御能力的生物标志物

Biomarkers of oxidative damage and antioxidant defense capacity in Caiman latirostris blood.

作者信息

Poletta Gisela L, Simoniello María Fernanda, Mudry Marta D

机构信息

Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje el Pozo, CP 3000, Santa Fe, Argentina; "Proyecto Yacaré"-Laboratorio de Zoología Aplicada: Anexo Vertebrados (FHUC-UNL/MASPyMA), Aristóbulo del Valle 8700, CP 3000, Santa Fe, Argentina; Grupo Investigación Biología Evolutiva (GIBE), IEGEBA-DEGE (CONICET-UBA), FCEyN, Pab. II, Ciudad Universitaria, Intendente Güiraldes 2160, CP 1428EGA, Buenos Aires, Argentina.

Cátedra de Toxicología, Farmacología y Bioquímica Legal, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Ciudad Universitaria, Paraje el Pozo, CP 3000, Santa Fe, Argentina.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2016 Jan;179:29-36. doi: 10.1016/j.cbpc.2015.08.003. Epub 2015 Aug 20.

DOI:10.1016/j.cbpc.2015.08.003
PMID:26299575
Abstract

Several xenobiotics, and among them pesticides, can produce oxidative stress, providing a mechanistic basis for their observed toxicity. Chronic oxidative stress induces deleterious modifications to DNA, lipids and proteins that are used as effective biomarkers to study pollutant-mediated oxidative stress. No previous report existed on the application of oxidative damage and antioxidant defense biomarkers in Caiman latirostris blood, while few studies reported in other crocodilians were done in organs or muscles of dead animals. The aim of this study was to characterize a new set of oxidative stress biomarkers in C. latirostris blood, through the modification of conventional techniques: 1) damage to lipids by thiobarbituric acid reactive substances (TBARS), 2) damage to DNA by comet assay modified with the enzymes FPG and Endo III, and 3) antioxidant defenses: catalase, superoxide dismutase and glutathione; in order to apply them in future biomonitoring studies. We successfully adapted standard procedures for CAT, SOD, GSH and TBARS determination in C. latirostris blood. Calibration curves for FPG and Endo III showed that the three dilutions tested were appropriate to conduct the modified comet assay for the detection of oxidized bases in C. latirostris erythrocytes. One hour of incubation allowed a complete repair of the damage generated. The incorporation of these biomarkers in biomonitoring studies of caiman populations exposed to xenobiotics is highly important considering that this species has recovered from a serious endangered state through the implementation of sustainable use programs in Argentina, and represents nowadays a relevant economic resource for many human communities.

摘要

几种外源性物质,其中包括农药,可产生氧化应激,为其观察到的毒性提供了机制基础。慢性氧化应激会导致对DNA、脂质和蛋白质的有害修饰,这些修饰被用作研究污染物介导的氧化应激的有效生物标志物。此前尚无关于凯门鳄血液中氧化损伤和抗氧化防御生物标志物应用的报道,而在其他鳄鱼中进行的少数研究是在死亡动物的器官或肌肉中进行的。本研究的目的是通过改进传统技术,在宽吻凯门鳄血液中表征一组新的氧化应激生物标志物:1)用硫代巴比妥酸反应性物质(TBARS)检测脂质损伤;2)用FPG和Endo III酶修饰的彗星试验检测DNA损伤;3)抗氧化防御:过氧化氢酶、超氧化物歧化酶和谷胱甘肽;以便将它们应用于未来的生物监测研究。我们成功地调整了宽吻凯门鳄血液中CAT、SOD、GSH和TBARS测定的标准程序。FPG和Endo III的校准曲线表明,所测试的三种稀释度适用于进行改进的彗星试验,以检测宽吻凯门鳄红细胞中的氧化碱基。一小时的孵育可使产生的损伤完全修复。考虑到该物种通过在阿根廷实施可持续利用计划已从严重濒危状态中恢复过来,并且如今是许多人类社区的重要经济资源,将这些生物标志物纳入接触外源性物质的凯门鳄种群的生物监测研究中非常重要。

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