Oxidative stress and DNA damages induced by cadmium accumulation

Experimental evidence shows that cadmium （Cd） could induce oxidative stress and then causes DNA damage in animal cells, however, whether such effect exists in plants is still unclear. In the present study, Vicia faba plants was exposed to 5 and 10 mg/L Cd for 4 d to investigate the distribution of Cd in plant, the metal effects on the cell lipids, antioxidative enzymes and DNA damages in leaves. Cd induced an increase in Cd concentrations in plants. An enhanced level of lipid peroxidation in leaves and an enhanced concentration of H2O2 in root tissues suggested that Cd caused oxidative stress in Vicia faba. Compared with control, Cd-induced enhancement in superoxide dismutase activity was significant at 5 mg/L than at 10 mg/kg in leaves, by contrast, catalase and peroxidaseactivities were significantly suppressed by Cd addition. DNA damage was detected by neutral/neutral, alkaline/neutral and alkaline/alkaline Comet assay. Increased levels of DNA damages induced by Cd occurred with reference to oxidative stress in leaves, therefore, oxidative stress induced by Cd accumulation in plants contributed to DNA damages and was possibly an important mechanism of Cd-phytotoxicity in Vicia faba plants.  

纳米材料对斑马鱼的氧化损伤及应激效应研究

以斑马鱼(Daniorerio)为受试动物,研究了纳米及常规TiO2、ZnO悬浮液对其鳃、消化道及肝脏的氧化损伤及应激效应,同时对纳米及常规TiO2、ZnO悬浮液中的颗粒形貌特征及·OH生成量进行了测定.结果发现,虽然纳米TiO2、ZnO颗粒与其常规颗粒在溶液中的粒径分布接近,但50mg/L纳米TiO2、ZnO悬浮液中·OH产生量(96h光照下,分别为2.17mmol/L、0.72mmol/L)远远高于50mg/L常规颗粒(未检测到).50mg/L纳米TiO2处理下,斑马鱼肝脏中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、还原型谷胱甘肽(GSH)、蛋白质羰基含量分别为对照的70.2%、65.4%、53%、178.1%;消化道中SOD活性及GSH、丙二醛(MDA)含量分别为对照的149.6%、212.9%、217.2%;鳃中MDA含量为对照的160.9%.而50mg/L常规TiO2悬浮液对斑马鱼没有产生毒理效应.5mg/L纳米及常规ZnO对斑马鱼肝脏的氧化伤害最强,其中5mg/L纳米ZnO处理组中SOD、CAT活性及GSH、MDA含量分别为对照的62.9%、53.1%、45.2%、204.2%,5mg/L常规ZnO处理组中SOD、CAT活性及GSH、MDA含量分别为对照组48.3%、51.8%、34.6%、289.6%;虽然斑马鱼鳃及消化道也受到明显氧化应激效应(p0.05),但并没有受到氧化损伤.研究表明,团聚作用对不同化学组成纳米颗粒的毒性影响程度不同;且不同化学组成纳米颗粒在生物体内可能通过不同机制产生了不同种类的ROS,从而对不同细胞组分产生的氧化损伤及应激效应是其重要的毒理机制.  

六价铬致小鼠DNA损伤及肝肾氧化应激的实验研究

为研究六价铬(Cr(Ⅵ))摄入后对机体造成的损伤,将重铬酸钾(K2Cr2O7)以25、50、100 mg·kg-1对小鼠进行灌胃,染毒时间为1 d、3 d和5 d.检测外周血淋巴细胞DNA损伤及肝肾组织活性氧自由基(ROS)水平、脂质过氧化产物丙二醛(MDA)含量,以及超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性.实验结果表明,经Cr(Ⅵ)染毒1 d、3 d和5 d后小鼠外周血淋巴细胞DNA损伤及肝ROS水平与对照组相比均有明显升高,肝脏抗氧化酶活性也有显著改变,但MDA含量差异不显著,而肾组织中各指标均未见有明显改变.由此认为,经口摄入Cr(Ⅵ)后能导致DNA损伤并且对肝脏造成氧化应激.  

纳米氧化锌对斑马鱼肝脏的毒性效应

采用半静态急性实验方法,研究斑马鱼在浓度为0、0.05、0.1、5、10、25、50 mg·L^-1的纳米氧化锌(nano-ZnO)中暴露4、24、96 h后,nano-ZnO对肝中超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)的影响,肝脏中Bcl-2、Bax、p53和MDM2的mRNA表达量的变化以及暴露7、15、30 d时肝组织解剖结构变化情况. 结果表明,与对照组相比,实验组表现为: ①肝组织出现组织水肿,部分细胞质空泡化、细胞核固缩;②肝巨噬细胞增多,窦间隙增大;③肝中SOD活性升高、MDA含量增加、CAT活性降低;④肝脏中Bax/Bcl-2和p53的mRNA的表达量均升高;⑤ MDM2 mRNA的表达量表现为在低浓度组中降低,高浓度组中则升高. 实验结果表明,nano-ZnO能引起斑马鱼肝脏发生氧化应激作用,使肝中抗氧化酶活性发生变化,并诱导肝脏中细胞凋亡相关基因的表达,使细胞发生凋亡且造成肝组织结构发生变化.  

Oxidative stress and apoptosis of Carassius auratus lymphocytes induced by nonplanar （PCB153） and coplanar （PCB169） polychlorinated biphenyl congeners in vitro

Among all the 209 kinds of polychlorinated biphenyls (PCBs) congeners, nonplanar and coplanar PCB congeners have di erent levels of toxicity on mammal cells such as neuronal cells, but little is known about their toxicity on fish cells although PCB congeners usually have high bioaccumulation abilities in the detected fish bodies. This study showed that 2,2’,4,4’,5,5’-hexacholorbiphenyl (PCB153, nonplanar congener) and 3,3’,4,4’,5,5’-hexacholorbiphenyl (PCB169, coplanar congener) caused apoptosis on the isolated crucian carp (Carassius auratus) lymphocytes and the induced cytotoxicity was structure-dependent. According to the laser confocal microscope observations, apoptosis was clearly distinguished by condensation of nucleus, shrinkage and formation of apoptotic bodies. DNA fragmentation was detected by agarose gel electrophoresis. These typical morphological and biochemical characteristics indicate the occurrence of apoptosis on fish lymphocytes. According to the flow cytometry analysis, after the cells were exposed to 10 mol/L PCBs for 3 h, the apoptotic percentage induced by PCB153 was 23.41%, while that induced by PCB169 was even higher (31.03%). Furthermore, incubating PCBs with fish lymphocytes enhanced levels of reactive oxygen species (ROS) and malondialdehyde (MDA), clearly indicating the presence of oxidative stress and lipid peroxidation. Our data also demonstrate that the di erent cytotoxic e ects induced by coplanar and nonplanar PCBs were correlated with their structural characteristics and the coplanar congener was more cytotoxic than nonplanar congener. This study suggests that cytotoxicity mechanisms of the PCB congeners on fish lymphocytes depend on their planarity and chemical structures.  

Effect of sub-acute exposure to TiO2 nanoparticles on oxidative stress and histopathological changes in Juvenile Carp （Cyprinus carpio）

Increasing application of nanotechnology highlights the need to clarify and understand nanotoxicity. Mammalian and in vitro studies have raised concerns about the toxicity of titanium dioxide nanoparticles (TiO₂-NPs), but there are limited data on ecotoxicity to aquatic organisms. In this work, the sub-acute toxicity of TiO₂-NPs to carp (Cyprinus carpio) was assessed. Superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities and lipid peroxidation (LPO) levels in liver, gill and brain tissues of carps varied with concentration of TiO₂-NPs suspensions and exposure time (up to 8 d). As a result, 100 and 200 mg/L TiO₂-NPs caused statistically significant decrease in SOD, CAT and POD activities and significant increase in LPO levels in tissues (P < 0.05), suggesting that the fish exposed to these two concentrations of TiO₂-NPs suffered from the oxidative stress. The extent of depletion of antioxidant enzymes activities and the elevation of LPO in the liver was the greatest, indicating that the liver might be the most susceptible organ to TiO₂-NPs exposure. In addition, carps had gill pathologies including edema and thickening of gill lamellae as well as gill filaments, and liver pathologies including necrotic and apoptosis hepatocytes after exposed to 100 and 200 mg/L TiO₂-NPs for 20 d. These results indicated a potential risk from TiO₂-NPs released into the aqueous environment.  

CdSe/ZnS量子点对稀有鮈鲫胚胎发育的影响及其氧化应激作用

本研究以稀有鮈鲫(Gobiocypris rarus)为对象,研究了不同浓度CdSe/ZnS量子点(QDs)暴露下,稀有鮈鲫胚胎发育过程中自主运动频率、内心率和体长的变化,以及利用体内超氧化物歧化酶(SOD)和丙二醛(MDA)作为毒性指标,反映CdSe/ZnS QDs暴露对稀有鮈鲫胚胎发育的氧化应激作用.结果显示:CdSe/ZnS QDs对稀有鮈鲫胚胎72 hpf(hours post fertilization)的半致死浓度(LC50)为319.629 nmol·L-1,96 hpf的半致畸浓度(EC50)为203.312 nmol·L-1.CdSe/ZnS QDs暴露不仅影响稀有鮈鲫胚胎死亡率、畸形率、自主运动频率、孵化时间和孵化率,而且使其内心率减缓、体长缩短,导致胚胎卵凝结,心包囊肿,出现脊椎弯曲等多种毒性现象.同时发现,CdSe/ZnS QDs暴露导致稀有鮈鲫体内MDA含量增加以及SOD活力的降低.这表明CdSe/ZnS QDs对稀有鮈鲫胚胎发育具有致畸、致死作用,而氧化应激可能是引起其胚胎致畸、致死的重要机制之一.  

Coking wastewater increases micronucleus frequency in mouse in vivo via oxidative stress

Coking wastewater has caused serious health risk in coal-producing areas of China, however its toxic effects have not been well understood. The genotoxicity induced by coking wastewater on mice in vivo and its possible oxidative mechanisms were investigated via observing the induction of micronuclei in polychromatic erythrocytes of mouse bone marrow, and subsequently determining the antioxidative enzyme activities （superoxide dismutase Cu, Zn-SOD, Se-dependent glutathione peroxidase, and catalase）, thiobarbituric acid reactive substance contents and protein carbonyl levels in brains and livers of mice. Results showed that the tested coking wastewater caused a significant increase of micronucleus frequencies in a concentration-dependent manner. Also, the sample increased lipid peroxidation and protein oxidation levels, which was accompanied by changes in antioxidative status. Interestingly, pre-treatment with an antioxidant （vitamin C） led to a statistical reduction in the micronucleus frequency caused by coking wastewater. This implies that coking wastewater induces evident genetic damage in mammalian cells, and exposure to polluted areas might pose a potential genotoxic risk to human beings; in the process, oxidative stress played a crucial role.  

土壤Pb暴露下赤子爱胜蚓(Eisenia fetida)的急性毒性和氧化应激反应

根据OECD试验指南,研究了人工土壤铅暴露下赤子爱胜蚓(Eisenia fetida)的急性毒性和氧化应激反应.结果表明:铅暴露对赤子爱胜蚓14 d的LC50(半致死浓度)为2 576.76 mg/kg,95%置信区间为2 388.42~2 792.00 mg/kg;铅暴露第1、7、14天赤子爱胜蚓的SOD(超氧化物歧化酶)活性,铅暴露第1、14天的POD(过氧化物酶)活性,铅暴露第14天的MDA(丙二醛)含量均与w(Pb)呈良好的剂量-效应关系,但CAT(过氧化氢酶)活性与w(Pb)未呈剂量-效应关系.赤子爱胜蚓体内3种抗氧化酶对人工土壤铅污染响应的敏感程度为SODPODCAT,SOD作为抗氧化系统的重要酶类可以更好地预测污染物对于赤子爱胜蚓的亚致死毒性.  

节球藻毒素诱导草鱼巨噬细胞氧化应激的机理研究

试验以鱼草(Ctenopharyngodon idellus)巨噬细胞为研究目标,进行体外诱导试验,研究节球藻毒素(nodularin,NOD)浓度变化对草鱼巨噬细胞的氧化应激和还原型谷胱甘肽(glutathione,GSH)解毒性能的影响。结果表明NOD会促进草鱼巨噬细胞内自由基产生,导致胞内脂质过氧化,并且在这过程中会抑制超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性,进一步促进巨噬细胞的氧化应激效应,致使活性氧簇(reactive oxygen species,ROS)水平和丙二醛(malondialdehyde,MDA)含量相比对照组分别升高2倍和3.8倍。此外,NOD抑制谷胱甘肽-S转移酶(glutathione S-transferase,GST)活性,降低GSH与NOD的结合能力。同时,降低谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)活性至对照组56%,提高谷胱甘肽还原酶(glutathione reductase,GR)活性至对照组3.3倍,促进GSH朝GSSG转变,从而达到降低GSH含量。因此,NOD会作用于鱼巨噬细胞,造成细胞内氧化应激加剧,GSH的解毒能力降低,最终导致巨噬细胞凋亡。