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纳米氧化锌对斑马鱼肠组织的氧化损伤
引用本文:刘林,赵群芬,朱帅旗,许峰.纳米氧化锌对斑马鱼肠组织的氧化损伤[J].水产学报,2015,39(11):1702-1711.
作者姓名:刘林  赵群芬  朱帅旗  许峰
作者单位:宁波大学海洋学院, 浙江宁波 315211,宁波大学海洋学院, 浙江宁波 315211,宁波大学海洋学院, 浙江宁波 315211,宁波大学海洋学院, 浙江宁波 315211
基金项目:水产学浙江省重中之重(一级)学科开放基金(xkzsc1415)
摘    要:为探讨纳米氧化锌颗粒(ZnO-NPs)对斑马鱼肠组织的损伤及损伤机理,研究了ZnO-NPs对肠组织结构、抗氧化酶活性及相关凋亡基因表达的影响。将斑马鱼分别暴露于浓度为0、0.05、0.1、5、10、25和50 mg/L的ZnO-NPs水体中,在4、24和96 h时,用比色法测定了肠组织中丙二醛(MDA)含量、谷胱甘肽过氧化物酶(GSH-PX)、谷胱甘肽硫转移酶(GST)活性;用实时荧光定量法测定了Bcl-2、Baxp53和MDM2 mRNA表达水平的变化;在第7、15和30天时用H.E染色技术观察了肠组织解剖结构变化。与对照组相比,实验组结果显示:斑马鱼肠中的MDA含量均高于对照组;随着时间的延长,组织中GSH-PX和GST活性呈先升高后下降的趋势;肠组织中Bax/Bcl-2和p53的mRNA的表达量均升高,而MDM2的mRNA表达量则随着时间的延长和浓度的增加表现为先降低后升高;H.E染色观察到肠组织结构发生变化,表现为杯状细胞增多,肿胀变形,部分细胞质空泡化,肠中淋巴细胞增多,肠绒毛侵蚀变形,且有时间和浓度依赖性。结果表明:ZnO-NPs能诱导斑马鱼肠组织产生氧化应激作用,使组织中抗氧化酶活性发生变化,诱导肠中细胞凋亡相关基因的表达,并且能对肠组织结构造成损伤。

关 键 词:斑马鱼  ZnO-NPs  肠组织  氧化损伤
收稿时间:5/4/2015 12:00:00 AM
修稿时间:2015/7/13 0:00:00

Oxidative damage of zinc oxide nanoparticles to zebrafish intestine
LIU Lin,ZHAO Qunfen,ZHU Shuaiqi and XU Feng.Oxidative damage of zinc oxide nanoparticles to zebrafish intestine[J].Journal of Fisheries of China,2015,39(11):1702-1711.
Authors:LIU Lin  ZHAO Qunfen  ZHU Shuaiqi and XU Feng
Institution:College of Marine Sciences, Ningbo University, Ningbo 315211, China,College of Marine Sciences, Ningbo University, Ningbo 315211, China,College of Marine Sciences, Ningbo University, Ningbo 315211, China and College of Marine Sciences, Ningbo University, Ningbo 315211, China
Abstract:To analyze the mechanism and result of the zinc oxide nanoparticles(ZnO-NPs) injury to zebrafish intestine tissue,this paper has studied the effect of ZnO-NPs on zebrafish intestine tissue structure,antioxidant enzyme activity and apoptotic gene expression.Zebrafish were exposed to the different concentrations of ZnO-NPs(0,0.05,0.1,5,10,25,50 mg/L).The malondialdehyde(MDA),glutathione peroxidase(GSH-PX) and glutathione-s-transferase(GST) in the intestine tissues of zebrafish were measured by colorimetric method at the 4th,24th and 96th hour.The mRNA expressions of Bcl-2, Bax,p53 and MDM2 in intestine tissues were also examined using quantitative real-time PCR assays.The change of anatomical structure in the intestine tissues of zebrafish was observed by Hematoxylin-Eosin(H-E)staining technique after zebrafish were exposed to ZnO-NPs for 7 d,15 d and 30 d.Compared with the control group,the experimental groups were shown as:the MDA content was increased;with a longer exposure time,the activity of GSH-PX and GST first increased and then decreased;the mRNA expression level of genes Bax/Bcl-2 ratio and p53 of experimental groups increased;with a longer exposure time and a higher concentration,the mRNA expression level of gene MDM2 first increased and then decreased.HE staining was observed in the experimental groups,changes in the structure of intestinal tissues were shown as the phenomena that the number of goblet cells and lymphocyte increased,goblet cells swelled,cytoplastic vacuolation,leukomonocyte increased and erosion of villi was observed in zebrafish intestine depending on the time and dose.The results suggest that the oxidative damage of ZnO-NPs to the zebrafish intestine was caused by the increase of oxidative stress,which made enzyme inactivating,induced the expression of cell apoptosis-related genes and caused the damage to histological structure of intestine.
Keywords:zebrafish  ZnO nanoparticles  intestine tissue  oxidative damage
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