镉、铜和锌胁迫下黑藻活性氧的产生及抗氧化酶活性的变化研究

研究了不同Cd、Cu、Zn处理浓度对黑藻体内活性氧()产生及对抗氧化酶(SOD、POD、CAT)活性的分子毒理学效应以探讨高等水生植物抗氧化酶对重金属胁迫的反应。结果表明,三种重金属都不同程度地加快了产生速率;Cu使SOD、POD、CAT活性下降;Cd也都减弱了SOD和POD活性,而CAT活性在0.5—5mg/L处理浓度时增加;Zn对SOD活性也为抑制作用,当浓度为0.5—5mg/L时POD和CAT活性都上升。关联度分析发现Cd、Cu和Zn胁迫下黑藻起主要保护作用的分别为SOD、POD和CAT,而SOD最易受到影响。Cd、Cu处理下的叶绿素含量也都呈下降趋势,而0.5—5mg/L的Zn浓度刺激了叶绿素合成。所有Zn处理、0.5mg/L的Cu处理和0.5—1mg/L的Cd处理的叶绿素a/b值都大于对照值。除了Cu使可溶性蛋白含量减少外,0.5—5mg/L的Zn和0.5—1mg/L的Cd都使其含量增加。综合起来,Cu的毒性最强,其次为Cd,Zn最弱。致死阈浓度分别为:Cu:0.5—1mg/L;Cd:1—2mg/L;Zn:5—6mg/L。SOD是评价重金属对沉水植物毒性效应的灵敏指标。黑藻对水环境Cu污染反应敏感。       

外源脱落酸对小麦幼苗抗镉胁迫能力的影响

以小麦(Triticum aestivum L.)为试材,采用水培法研究了100mg/L镉(Cd2+)胁迫条件下施用外源脱落酸(ABA)对小麦幼苗生长及某些生理生化指标的影响。结果表明:(1)100mg/L Cd2+胁迫下,小麦叶片膜脂过氧化产物丙二醛(MDA)含量显著提高,植株生长受到抑制;(2)外源ABA能够明显提高Cd2+胁迫小麦幼苗的根系活力,增加其叶片SOD、CAT和POD活性,促进其脯氨酸积累,降低MDA的含量,并以5.0μmol/L ABA的效果最明显;(3)1.0～5.0μmol/L外源ABA不同程度地缓解Cd2+胁迫对小麦幼苗生长的抑制作用,且5.0μmol/L时效果最明显,其株高、根长、总干重分别比单一Cd2+胁迫处理显著提高6.73%、149%和10.52%,而10.0μmol/LABA反而加重了Cd2+对小麦幼苗生长的伤害。因此,适宜浓度的外源ABA能够通过增加体内保护酶活性和脯氨酸含量来缓解Cd2+胁迫对小麦幼苗生长的抑制作用,增强小麦幼苗的抗Cd2+胁迫能力,并以5.0μmol/L ABA处理效果最好。     

重金属Cu2+、Pb2+单因子及联合毒性对泥鳅卵细胞DNA的损伤效应

采用室内暴露试验方法, 研究了不同浓度Cu2+(0.01、0.10、0.25mg/L)、Pb2+(0.05、0.50、0.75mg/L)单因子染毒以及Cu2++Pb2+(0.01 mg/L+0.05mg/L、0.10 mg/L+0.50 mg/L、0.25 mg/L+0.75 mg/L)联合染毒对泥鳅卵细胞DNA的损伤效应, 并以SCGE技术进行检测。结果显示, Cu2+与Pb2+单因子染毒对泥鳅卵细胞DNA的损伤具有较为显著的剂量-效应与时间-效应关系(P0.05)。Cu2++Pb2+联合染毒, 在溶液暴露的0-5d表现为剂量-效应与时间-效应关系(P0.05); Cu2++Pb2+暴露5-10d 则表现出拮抗作用。研究结果显示, Cu2+、Pb2+ 单因子及联合染毒均造成泥鳅卵细胞DNA损伤, 具有基因毒性效应。       

镉对尖紫蛤抗氧化酶活性及脂质过氧化的影响

为阐明镉(Cd2+)对尖紫蛤消化盲囊和鳃抗氧化酶的毒性影响程度,研究了不同浓度的Cd2+(0.005、0.05、0.5 mg/L)在不同暴露时间(24h、72h、120h)对尖紫蛤鳃和消化盲囊中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)的活性以及丙二醛(MDA)含量的影响。结果表明,在Cd2+浓度为0.005 mg/L时,在整个实验期间Cd2+对消化盲囊和鳃内的SOD、CAT、GSH-PX活性并无显著影响。在Cd2+浓度为0.05 mg/L和0.5 mg/L时,SOD、CAT、GSH-PX在鳃和消化盲囊中的活性都呈现出明显的时间剂量依赖关系。在0.05 mg/L暴露时,鳃和消化盲囊中的SOD、CAT和GSH-PX的活性随时间逐渐增强,在72h时达到最大值,但在120h时略有降低。在0.5 mg/L暴露时,消化盲囊中SOD、CAT及鳃中CAT活性在24h时上升达到最大值,但鳃中SOD直到72h才达到最大值,并均在120h下降到最低,其中消化盲囊中SOD和CAT活性在120h低于对照组,这可能与消化盲囊对Cd2+的敏感性高于鳃有关。在0.5 mg/L暴露的鳃中,GSH-PX在24h、72h活性并不上升,在120h甚至低于正常水平,0.5 mg/L暴露的消化盲囊中,24h时迅速增高,然后逐渐下降到正常值。这可能与Cd2+结合了GSH-PX的活性中心,降低了GSH-PX的活性有关。与3种酶活性随着时间延长和剂量的增加,酶活性会降低的变化趋势不同,鳃和消化盲囊中的MDA的含量随时间延长和剂量的增加而增加,并不出现下降,这表明尖紫蛤鳃和消化盲囊中的MDA含量可以灵敏的反映机体内的氧化损伤程度,但不能敏感的反映水体中Cd2+的污染情况。     

重金属镉和铅胁迫对海洋微藻的毒性效应研究

设置不同浓度的重金属Cd2+(0、0.2、0.4、0.6、0.8和1 mg•L-1)和Pb2+(0、0.1、0.2、0. 4、0.8和1.6 mg•L-1)胁迫处理, 检测米氏凯伦藻(Karenia mikimotoi)的生长生理情况, 分析重金属胁迫对海洋微藻生长的毒性效应, 探讨超氧化物歧化酶(SOD)和过氧化氢酶(CAT)等对重金属胁迫的缓解作用。研究结果发现, 重金属Cd2+和Pb2+对米氏凯伦藻具有较强的毒性, 随着重金属浓度的提高, 细胞生长受到毒害作用增强; 而米氏凯伦藻对重金属Cd2+和Pb2+胁迫具有一定的适应性。重金属Cd2+和Pb2+胁迫导致米氏凯伦藻细胞的叶绿素a、叶绿素b含量下降(1.6 mg•L-1 Pb2+胁迫除外), 类胡萝卜素含量提高, 最大光能转化效率(Fv/Fm)下降, 表明重金属胁迫抑制藻的光合作用, 影响藻的生长繁殖。丙二醛(MDA)随着重金属Cd2+和Pb2+浓度提高而升高, 说明重金属引起藻细胞膜透性增加, 藻细胞遭受破坏。SOD活性整体呈现先升高后下降(或与对照持平)的趋势, 提示海洋微藻的抗氧化酶系统在低浓度重金属胁迫下产生应激性反应, 酶活性增强; 而CAT活性上升, 也对藻细胞起到保护作用, 推测两者共同反应, 以缓解藻体遭受的重金属毒害作用。结果可为了解重金属胁迫对海洋微藻的毒性效应提供参考。     

镉对克氏原螯虾肝胰腺抗氧化系统的影响

采用毒性试验方法,研究了不同浓度Cd2+(1.72、3.44、6.89、13.77和27.55mg·L-1)对克氏原螯虾肝胰腺抗氧化酶(SOD、CAT、GSH-PX)和抗氧化物质(GSH、Vc)的影响.结果表明:超氧化物歧化酶(SOD)活性受低浓度Cd2+诱导和高浓度Cd2+抑制,且受抑制程度与Cd2+浓度呈正相关.过氧化氢酶(CAT)活性总体表现为先激活后下降,且在第3天达最大值,CAT活性对低浓度Cd2+(≤6.89 mg·L-1)暴露敏感,而高浓度Cd2+对其影响较小.谷胱甘肽过氧化物酶(GSH-PX)活性对Cd2+浓度敏感,当Cd2+≤6.89 mg · L-1时,GSH-PX活性先升高后下降,更高浓度下GSH-PX活性在暴露后第1天即表现出抑制作用.还原型谷胱甘肽(GSH)含量在Cd2+≤6.89 mg·L-1时始终被诱导,且均在第1天达最大值,而高浓度Cd2+(≥13.77 mg·L-1)对GSH含量影响不明显.维生素C(Vc)对Cd2+胁迫敏感,各处理组Vc含量在第1天均显著下降,且下降程度与Cd2+浓度呈正相关,之后具有一定的合成恢复能力.抗氧化酶和非酶抗氧化物质在抵御Cd2+胁迫反应中共同发挥作用,且大多表现出明显的时间和剂量效应性.GSH-PX和Vc可以作为Cd2+污染的潜在生物指示物.     

匍茎翦股颖对Cu2+、Zn2+、Cd2+与Pb2+ 胁迫的生长响应与阈限浓度

采用砂培法,研究了匍茎翦股颖对Cu2+、Zn2+、Cd2+与Pb2+胁迫的生长响应及阈限浓度,结果表明:种子萌发率随着4种重金属浓度的增加而下降。对株高的影响是当重金属浓度小于100mg/L时会促进株高生长,高于100mg/L则产生抑制作用。Cu2+显著抑制根系生长,并随浓度的增加抑制效应愈加显著;在Cu2+浓度为600mg/L时匍茎翦股颖的根长比对照下降了93.75%。Cu2+、Zn2+、Pb2+浓度小于200mg/L时会促进地上生物量的增加,但高于200mg/L时,地上生物量会随着3种重金属的增加而减少。Cu2+、Zn2+浓度小于100mg/L或Cd2+、Pb2+浓度小于200mg/L会增加叶绿素的含量,高浓度会降低叶绿素的含量;Cd2+在浓度为600mg/L时显著降低叶绿素含量,与对照相比,下降了43.55%。匍茎翦股颖生长的综合效应分析表明,匍茎翦股颖对Cu2+胁迫最敏感,具有较低的阈限浓度,而Zn2+胁迫对匍茎翦股颖的生长影响最小,阈限浓度相对较高。     

Cd2+胁迫下浮萍叶细胞核超微结构、nDNA损伤及抗氧化酶系的变化

以不同浓度Cd2+处理6d的浮萍为材料,从细胞核超微结构损伤、nDNA一级结构变化及抗氧化酶系活性改变等方面分析重金属的植物毒理学效应。透射电镜观察发现,5～7mg/L Cd2+处理的细胞核膜完整,染色质凝聚并趋边化;10mg/L Cd2+处理后则核仁解体,且DNA原位末端标记(TUNEL)检测发现DNA的3′-OH断端可被特异标记,表现出典型的凋亡细胞特征。而20mg/L Cd2+已导致细胞坏死。同时,较低浓度的Cd2+胁迫可刺激抗氧化酶(SOD,POD,CAT)活性升高,以清除体内活性氧,而随金属浓度的进一步增高,三种抗氧化酶活性都急速下降。研究发现,活性氧和抗氧化酶系密切参与了重金属Cd2+诱导的浮萍体细胞凋亡过程。     

镉、汞暴露在中华绒螯蟹卵巢中的富集及其氧化应激反应

研究以中华绒螯蟹(Eriocheir sinensis)为实验对象, 用不同浓度的Cd2+ 溶液(0、0.63、1.26、2.52、5.04和10.07 mg/L)或Hg2+ 溶液(0、0.06、0.11、0.23、0.46和0.91 mg/L)对其分别进行染毒, 6d后解剖取其卵巢, 检测卵巢中Cd和Hg的富集量以及氧化应激相关指标的变化情况。结果显示, 随着Cd2+或Hg2+染毒浓度的升高, 卵巢中Cd和Hg的富集量显著增加; H₂O₂、MDA和NO含量、MT mRNA表达量及NOS活性均随着Cd2+和Hg2+浓度的增加出现上升的趋势, 但SOD、CAT和GPx活性呈先升后降的趋势。这说明镉、汞均可引起卵巢组织产生过量的自由基和脂质过氧化, 并诱导氧化应激酶及MT mRNA的表达以抵抗不良的环境。总之, MT对中华绒螯蟹卵巢中重金属的解毒发挥着重要作用, 抗氧化酶能有效地消除活性氧自由基(ROS), 维持机体的氧化还原处于平衡状态, 共同保护膜脂免受氧化损伤。     

重金属对绿球藻硝酸还原酶活性的影响

研究4种重金属离子(Cu2 、Cd2 、Zn2 、Pb2 )对绿球藻硝酸还原酶(NR)活性的影响。4种重金属离子的实验浓度分别为0.01、0.1、1、10、50、100、200 mg/L;0.1、1、5、10、50、100、200 mg/L;0.1、1、5、10、50、100、200 mg/L;0.1、1、10、50、100、200、400 mg/L,BG11培养基作对照。研究结果表明:Cu2 、Cd2 分别在低浓度0.01、0.1 mg/L时,便开始抑制硝酸还原酶活性,随着Cu2 、Cd2 浓度增加,抑制作用增强,酶活性逐渐降低;当Cu2 、Cd2 ≥10 mg/L时,NR活性检测不到。当Zn2 、Pb2 浓度分别低于0.1 mg/L和50 mg/L时,NR活性随着培养基中Zn2 、Pb2 浓度的增大逐渐增强;当Zn2 、Pb2 浓度分别大于0.1 mg/L和50 mg/L时,NR活性逐渐降低。结果提示:从4种重金属离子对NR活性的影响看,绿球藻对Zn2 、Pb2 耐受力高,特别是Pb2 ,而对Cu2 、Cd2 的耐受能力要低些。     

镉胁迫对大弹涂鱼肝脏黄螵呤氧化酶和抗氧化酶活性的影响

研究了不同浓度镉离子对大弹涂鱼肝脏黄嘌呤氧化酶（XOD）、抗氧化酶（超氧化物岐化酶SOD、过氧化氢酶CAT）活性和丙二醛（MDA）含量的影响,以探讨其用于污染暴露的生物标记的可行性.结果表明,低浓度Cd²⁺（0.05 mg·L^-1）暴露使大弹涂鱼肝脏XOD和SOD活性随时间延长升高,第10天达到最大值,中高浓度暴露（0.5 和5 mg·L^-1 Cd²⁺）XOD和SOD活性显著或极显著升高;低和高浓度镉胁迫处理的CAT活性在12 h显著降低,随时间的延长低浓度组CAT活性恢复正常,高浓度组在第7天降到最低值, 并在恢复期的5 d中高浓度组CAT活性却极显著升高;低和中浓度镉胁迫处理的MDA含量12 h极显著升高,而高浓度却极显著下降,随时间延长低浓度恢复正常, 中浓度先上升后下降并到第5天达到最大值,而中高浓度在恢复5 d后MDA含量都极显著降低.     

Translation of muscle mRNA in rats following acute exposure to Pb2+ or Cd2+

The hindleg muscle of rats was studied 2 days following the i.p. administration of 0.5 mg Pb2+ /100 g body wt or 0.12 mg Cd2+ /100 g body wt or both Pb2+ and Cd2+. The incorporation of [14C]leucine into proteins was measured using mRNA obtained from the muscle polysomes. The translatability of this poly(A)+ RNA in a mRNA-dependent reticulocyte lysate was elevated similarly in each of the preparations from heavy metal treated rats compared to control rats. Evidence for increased mRNA activity for glyceraldehyde-3-phosphate dehydrogenase and actin was obtained.     

Response of catalase activity to Ag+, Cd2+, Cr6+, Cu2+ and Zn2+ in five tissues of freshwater fish Oreochromis niloticus

Catalase (CAT, EC 1.11.1.6) is an important enzyme in antioxidant defense system protecting animals from oxidative stress. Freshwater fish Oreochromis niloticus were exposed for 96 h to different concentrations of Ag(+), Cd(2+), Cr(6+), Cu(2+) and Zn(2+), known to cause oxidative stress, and subsequently CAT activities in liver, kidney, gill, intestine and brain were measured. In vivo, CAT was stimulated by all metals except Ag(+) in the liver and the highest increase in CAT activity (183%) resulted from 1.0 mg Cd(2+)/L exposure, whereas 0.5 mg Ag(+)/L exposure resulted in a sharp decrease (44%). In tilapia kidney, cadmium and zinc had no significant effects on CAT activity, whereas 0.1 mg Cr(6+)/L exposure caused a decrease (44%). Cadmium and zinc did not significantly affect the CAT activity in gill; however, 0.5 mg Ag(+)/L exposure caused an increase (66%) and 1.5 mg Cr(6+)/L exposure caused a decrease (97%) in CAT activity. All metals, except Cu(2+)(41% increase), caused significant decreases in CAT activity in the intestine. In brain, 1.0 mg Zn(2+)/L resulted in an increase in CAT activity (126%), while 1.5 mg Ag(+)/L exposure caused a 54% decrease. In vitro, all metals -- except Ag(+) and Cu(2+) in kidney -- significantly inhibited the CAT activity in all tissues. Results emphasized that CAT may be considered as a sensitive bioindicator of the antioxidant defense system.     

Effects of heavy metals Cd2+, Pb2+ and Zn2+ on DNA damage of loach Misgurnus anguillicaudatus

The effects of heavy metals Cd2+, Pb2+ and Zn2+ at 0.05, 0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure were examined by single cell gel electrophoresis (SCGE). For each test group, 20 loaches with similar body size (5.17-7.99g; 11.79-13.21 cm) were selected and kept in aquaria with dechlori-nated water at (22±1)℃ and fed a commercial diet every 48 h. According to the percentage of damaged DNA with tail and its TL/D (tail length to diameter of nucleus) value, the relationship between DNA damage degree and heavy metal dose and exposure time was determined. Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time. The highest percentage (84.85%) of damaged DNA was shown in 5.0 mg/L Zn2+ group after 28 days exposure and the biggest TL/D value (2.50) in all treated groups after 35 days exposure. During the first treated week, the damnification of DNA was mainly recognized as the first level, after that time, the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%. The joint toxic effects among Cd2+, Pb2+ or Zn2+ revealed much complexity, but it generally displayed that the presence of Cd2+ could enhance the genotoxicity of Pb2+ or Zn2+. In conclusion, the results suggested that there was a significant time-and dose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach, and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms.     

镉胁迫对萝卜幼苗活性氧产生、脂质过氧化和抗氧化酶活性的影响

镉胁迫使萝卜幼苗超氧阴离子(O2)、过氧化氢(H2O2)和丙二醛(MDA)含量增加;随着镉浓度提高,超氧化物歧化酶(SOD)活性首先明显上升,然后逐渐下降,甚至低于对照;叶片过氧化氢酶(CAT)活性明显增加,根系CAT活性则减少;根系以及较高浓度镉处理后期叶片的谷胱甘肽还原酶(GR)活性均显著增加.由此推测:在胁迫初期可能主要由SOD和CAT发挥抗氧化作用,而在胁迫后期由于抗坏血酸-谷胱甘肽(AsA-GsH)循环途径的激活,还原型谷胱甘肽和植物络合素含量的提高可能在清除活性氧或者直接螯合镉中起作用.     

Cd2+处理对菹草叶片保护酶活性和细胞超微结构的毒害影响

以不同浓度Cd^2 处理5d的菹草为实验材料,测定了叶片SOD,POD,CAT等生理生化指标的变化,并用透射电镜观察了Cd^2 对叶细胞超微结构,尤其是对叶绿体,线粒体和细胞核的损伤情况。结果表明：SOD活性,叶绿素含量随Cd^2 处理浓度的增加而下降,而CAT和POD活性都是在1mg／L浓度下达到峰值,而后降低。SOD对Cd^2 毒害最敏感,其次为POD和CAT。电镜观察发现：随Cd^2 浓度的增加,对细胞超微结构的损伤程度也加剧。表现为叶绿体膨大,被膜断裂、消失和叶绿体解体;线粒体变形,脊突膨大和空泡化;细胞核核仁分散,核膜断裂,核空泡化。并探讨了Cd^2 对植物的毒害机制。     

铅和镉污染对大羽藓生理特性的影响

This paper dealt with the effects of ²⁺,²⁺ and their combined pollution on the contents of chlorophyll,potassium and calcium in Thuidium cymbifolium.The results showed that except at 0.1 mg ²⁺稬^-1,the chlorophyll content decreased with increasing ²⁺ and ²⁺ concentrations,which was 18% of the control at 100 mg ²⁺稬^-1,and decreased by 48.6% at 200 mg ²⁺稬^-1.The potassium and calcium contents also decreased with increasing pollutants concentrations,being decreased by 61.1% at 100 mg ²⁺稬^-1.²⁺ had a stronger toxicity than ²⁺,and the toxicity of their combined pollution was stronger than that of each pollutant.²⁺ could increase the toxicity of ²⁺.     

Effects of zinc and cadmium on erythrocyte antioxidant systems of a freshwater fish Oreochromis niloticus

In this work to determine the effects of metals exposure of Oreochromis niloticus on erythrocyte antioxidant systems, fish were exposed to 5.0 mg/L Zn, 1.0 mg/L Cd, and 5.0 mg/L Zn + 1.0 mg/L Cd mixtures for 7 and 14 days and reduced glutathione (GSH) level, catalase (CAT), and glucose‐6‐phosphate dehydrogenase (G6PD) activities were investigated. In addition, Zn or Cd levels in whole blood were studied. Erythrocyte GSH level and CAT and G6PD enzyme activities increased in response to single and combined Zn and Cd exposure. The elevation observed in the CAT activity was higher in the Cd alone, and in combination with Zn, than in Zn alone. Time‐dependent alteration was not observed in all antioxidant parameters. Exposure to metals (alone and in mixture) resulted in elevatation of Zn and Cd levels in the blood. Concentration of metals in the blood of fish exposed to the Zn + Cd combination was lower than in fish exposed to the single metal. This study demonstrates that metals caused oxidative stress in fish erythrocytes, and an adaptation with an increase in CAT and G6PD activities and GSH level, which were important in the protection against metal damage, was observed. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 24:223–229, 2010; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.20327