石油烃对沙蚕镉生物富集特性及金属硫蛋白诱导的影响

多毛类沙蚕作为海陆交错带的关键性物种,常被作为环境监测的指示生物.我国海陆交错带沙蚕常见种——双齿围沙蚕经常遭受各种污染物\[包括重金属镉(Cd)和石油烃\]的胁迫.本研究采用慢性微宇宙试验,以暴露于单一和复合Cd和石油烃条件下的沙蚕为研究对象,重点考察石油烃对沙蚕Cd生物富集特性及诱导金属硫蛋白的影响.结果表明: 单一Cd胁迫条件下,沙蚕对重金属Cd的生物累积随暴露浓度的升高而显著上升,生物富集系数随暴露时间的延长呈上升趋势;复合胁迫条件下,与对照组相比,石油烃的添加可以显著增加沙蚕对重金属Cd的生物累积.Cd可以诱导沙蚕金属硫蛋白(MT)的表达,但当沙蚕体内Cd含量达到180 mg·kg^-1DM时,MT基本饱和;石油烃不能显著诱导沙蚕MT合成,与单一Cd处理相比,石油烃的添加能够显著影响Cd诱导的MT含量.这说明尽管石油烃本身不能直接影响MT合成,但在与Cd共存时可以调节MT的表达,因此在利用野生沙蚕MT作为监测指标时要具体区分蛋白表达的潜在诱导因素.     

厦门海域两头中华白海豚体内微量元素的积累

对2004年4月在厦门海域搁浅死亡的两头中华白海豚（Sousa chinensis）进行了肝脏、胰、胃、肾、肠、肺、肌肉、心脏等多组织微量元素锌（Zn）、铜（Cu）、汞（Hg）、铅（Pb）、镉（Cd）和砷（As）浓度的测定。两豚各组织总体上必需元素Zn、Cu浓度高于有毒元素Hg、Pb、Cd和As。XM20040430体内特别高的Zn、Hg、Pb、Cd浓度分别出现于皮肤（500μg/g干重）、肝脏（255μg/g干重）、肺（13.0μg/g干重）和肾（2.82μg/g干重）中,而Cu和As在各组织中的浓度较为相似。XM20040429体内仅肝脏中Zn（448μg/g干重）和Cu（52.0μg/g干重）的浓度远高于其他组织。虽然两海豚微量元素的总体积累多低于急性毒性水平,但有毒元素Hg、Pb、Cd、As的明显检出和XM20040429肝脏中异常高的Zn浓度均显示出进一步研究微量元素慢性胁迫效应的必要性。结果还显示厦门港海域近年来Hg、Cd、Pb污染未见减轻,As也明显检出,必须引起高度重视。今后有关水生哺乳动物中As积累的研究也有待加强。     

牛组织重金属含量与饲养环境的相关性

通过选择不同污染程度的区域进行取样,分析了贵州地区不同污染水平下牛组织中重金属(Cu、Zn、Pb、Cd)的含量和与饲养环境的相关性.结果表明:贵州不同污染水平下牛组织中的Cu、Zn基本符合国家食品卫生要求,Pb只有污染地区的肝脏和肾脏超过限量标准,Cd污染较严重,除非污染区肝脏外,牛肾脏和肝脏中Cd的平均含量均超过国家肉类制品卫生限量标准,但肌肉组织符合卫生标准;牛组织重金属元素含量与饲养环境中的土壤、饲料和饮用水源的重金属含量和污染程度密切相关,尤其是肾脏组织,其相关系数r＞0.78.饲料向牛组织的重金属迁移系数,随饲料元素含量的增加而逐渐降低,其中Cd的迁移系数最大,Pb的迁移系数最低;必需元素和有害元素在不同组织中的比值,随污染程度的增加而降低.肾脏的Cu/Cd和Zn/Cd值比其它组织低得多,Cd主要在肾脏中蓄积,Cu主要在肝脏组织蓄积,Zn主要在肌肉和肝脏,Pb主要在肾脏和肝脏.     

重金属在土壤生态系统中的富集及毒性效应

对清水塘重金属污染区土壤生态系统重金属的富集及毒性效应的研究结果表明,土壤中 主要富集的重金属元素为生物毒性较强的Cd、Hg、As以及具有一定毒性的Zn、Pb、Cu,平均综 合污染指数为640.大型土壤动物重金属含量的分析结果表明,重金属富集量随污染程度的 增加而增加,尤其某些巨蚓科种类对Cd表现高富集,富集系数为1196.通过染毒试验进一 步证明,土壤动物种类和数量随Cd浓度的增加而减少.同工酶分析表明,Cd对蚯蚓同工酶活性 有明显的毒性效应.污染区主要种植蔬菜作物,而重金属在蔬菜体内的富集量也随污染 程度的增加而增加,其中蔬菜对Cd的富集有明显的品种间差异.     

公路绿化植物油松(Pinus tabulaeformis)和小叶杨(Populus simonii)对重金属元素的吸收与积累

对内蒙古西部公路绿化植物油松（Pinus tabulaeformis）、小叶杨（Populus simonii）及其根际土壤中重金属元素（Cd、Hg、Pb、Cu、Zn、Ni、Cr）和类金属元素（As和Se）含量以及根际土壤重金属（Cu、Zn、Pb、Ni和Cr）形态、土壤pH值进行了测定。对比分析了公路沿线不同绿化植物及其不同器官对重金属元素的吸收与积累特征。结果表明：绿化植物根际土壤对重金属元素的吸附及污染程度以Cd为最高。随原子序数的递增,小叶杨和油松两种植物的根部和茎叶两种营养器官中重金属的含量均表现出“N”字形变动趋势。而且重金属元素在不同植物不同器官中的含量具有Zn〉Cu〉Ni,Cr,As,Pb〉Cd〉Hg的基本规律。小叶杨茎叶对重金属元素Cr、Ni和Pb的富集能力较根部为强,油松茎叶对重金属元素Cr、Ni、Cu和Pb的富集能力较根部为强。绿化植物根际土壤重金属元素有效态占总量百分比的大小序列为Zn〉Pb〉Ni、Cr〉Cu,与重金属元素在不同植物不同器官中的含量大小序列Zn〉Cu〉Ni、Cr、As、Pb〉Cd〉Hg并非趋于一致。公路绿化植物对根际土壤中重金属元素的吸收和积累与重金属元素有效态所占的比例有关。     

不同因素诱导豚鼠肝中金属硫蛋白的研究

通过皮下注射的方法诱导豚鼠产生金属硫蛋白（ＭＴ）,研究了重金属元素（Ｃｄ）、微量元素（Ｃｕ,Ｚｎ）及有机试剂（ＣＣｌ４,在体内可产生自由基）等因素的诱导与豚鼠肝脏中ＭＴ不同亚型的含量及金属结合状态的变化关系．实验结果表明,微量元素及有机试剂的诱导可使豚鼠肝脏中ＭＴ１的产量明显高于ＭＴ２,说明在体内ＭＴ１在参与微量元素的储存及清除自由基功能方面比ＭＴ２强．在重金属元素诱导下体内ＭＴ１对重金属元素的结合量远远大于ＭＴ２．表明ＭＴ１的重金属解毒能力比ＭＴ２强．上述实验结果与对不同亚型ＭＴ生物学功能差异的体外研究结果相吻合．此外,无论采用上述何种因素诱导,所得ＭＴ中均结合有Ｃｕ．对Ｃｕ在ＭＴ形成过程中的作用也进行了初步探讨．     

不同活动、饥饿和饮食成分状态下FGF21的动力学表达

成纤维细胞生长因子21（fibroblast growth factor,FGF21）是FGF家族中的新成员．目前研究显示,FGF21是一个新的糖脂代谢调节因子,有望成为治疗糖尿病的新型药物．为探讨FGF21的生理功能,利用real-time PCR和Western印迹,检测FGF21在不同生理或病理状态下基因水平和蛋白水平的表达量变化规律．实验结果显示,在全天24 h中,小鼠肝脏中FGF21在晚18点至21点,表达量显著升高,这可能与啮齿类动物傍晚活动加强及进食习性有关|FGF21在饥饿后表达量显著升高,在饥饿后喂食FGF21的表达量下降,并且随着饥饿时间的延长,FGF21的表达量升高,说明FGF21与饥饿程度呈正相关|灌注葡萄糖后20 min内,FGF21的表达量下降,而灌注脂肪乳20 min内,FGF21的表达量上升,说明葡萄糖是FGF21的负调节因子,而脂肪乳是FGF21的正调节因子|利用谷氨酸钠造模的肥胖小鼠,肝脏中FGF21的表达量显著高于同龄对照组,说明肥胖可诱导FGF21高表达．综上所述,FGF21的表达量变化与小鼠夜间活动取食、饥饿程度、饮食中不同的成分以及肥胖有关.     

重金属复合污染对小麦种子根活力的影响

应用二次回归旋转设计法,以小麦种子为材料,研究了水培条件下重金属复合污染对其根生长及活力的影响;借助回归分析建立了相应的数学模型,分析了Cu、Zn、Cd和Pb4种重金属对根活力的毒性、两两重金属问的联合作用以及各重金属的边际毒性效应．结果表明重金属对根活力的影响顺序为Pb＞Cu＞Cd＞Zn;Cu与Cd、Cu与Zn、Zn与Cd以及Cd与Pb元素间的互作效应显著;在0-20mg.L^-1浓度范围内,Cu和zn两元素在最小浓度时的边际效应值最大,Cd和Pb两元素的边际效应最大值却出现在最高浓度处．     

不同营养状态下方斑东风螺的镉生物积累及毒性

采用室内模拟方式,研究了方斑东风螺在饥饿及摄食饵料分别为贻贝和沙蚕3种情况下暴露于水体镉(Cd2+,50 μg·L-1)10周后的存活、生长、螺体Cd蓄积及有关毒性效应参数的变化.结果表明:长期饥饿与同时水体Cd暴露降低了方斑东风螺的成活率,且螺体糖原被最大程度动用,摄食有助于螺体抵抗Cd毒性;方斑东风螺肝胰脏DNA受损伤后在摄食状态下随时间延长均有不同程度的自我修复能力,但长期禁食使螺的DNA完整性降低且不能恢复;饥饿引起方斑东风螺体组织萎缩,并导致Cd蓄积与金属硫蛋白(MT)含量升高,摄食螺因体质量增加的稀释作用而具有较低的Cd蓄积与MT含量;摄食贻贝的方斑东风螺较摄食沙蚕有更快的生长率和更低的Cd浓度.表明饥饿加剧了Cd对方斑东风螺的毒性;不同饵料对方斑东风螺的生长有显著影响,并间接影响螺体Cd蓄积、MT诱导和糖原消耗.应用方斑东风螺等海洋腹足类作为环境监测指示种时应考虑栖息地食物丰度和饵料类型等的影响.方斑东风螺高密度工厂化养殖中应注意合理投饵并定期监测海水Cd浓度.     

行道树毛白杨树干中重金属元素分布

采用电感耦合等离子体发射光谱法(ICP),测定分析首都机场高速公路旁毛白杨(Populus tomentosa Carr.)树干中Pb、Cd、Cr、Cu、Zn、Ni和Mn 7种重金属元素的含量及积累量,比较分析树干不同组织、不同方位、不同龄级年轮重金属含量差异及与交通量、关键气候因子的相关性。结果表明:树干木质部中各重金属平均含量由大到小依次为Zn、Cu、Cr、Mn、Ni、Pb、Cd,树皮中依次为Zn、Mn、Cr、Pb、Cu、Cd、Ni,树皮中各重金属元素的含量明显高于木质部;同一树干木质部中,各重金属元素在不同方位的分布有所差异,其中,靠近车道一侧的各重金属元素含量均高于背离车道一侧,南北向比较中,Ni、Zn为南侧含量高于北侧,其他5种重金属元素均为北侧高于南侧;以5a为一个龄级将年轮划分为5个龄组,各龄级年轮中重金属含量随时间的变化趋势各异,其中Pb、Cd、Cu、Zn总体呈递减趋势,且与年降水量、最低气温、日照时数、雨天日数和大风日数呈正相关趋势,与年平均气温和最高气温呈负相关趋势;各元素在毛白杨树干木质部中的积累量表现为ZnCrCuMnPbNiCd。     

Induction and degradation of Zn-, Cu- and Cd-thionein in Chang liver cells

Human liver cells (Chang liver) were exposed to 5 micrograms Zn, 2.5 micrograms Cu or 1 microgram Cd/ml in cultured medium. These exogeneous heavy metals were accumulated by the cells and induced de novo synthesis of metallothionein after a 3-h incubation period. The production of Zn-, Cu- or Cd-thionein started in the cells with accumulation of 1 nmol Zn, 0.3 nmol Cu and 0.1 nmol Cd/mg cytosol protein and subsequently the amounts of metal-binding thioneins increased in agreement with the relative amount of metal accumulated in the cytosol over a 24-h period. When cells containing Zn- or Cu-thionein were placed in metal free medium, 70% or 25% of the zinc or copper bound to each original metallothionein was released after 3 h; bound metals decreased to 85% and 65% respectively after 24 h. The disappearance of metal from metallothionein correlated with increases of metal in the medium. On the other hand, 35S-counts incorporated into Zn- and Cu-thionein decreased only to 40% and 15% of the levels in the original metallothionein after 3 h; 35S-counts decreased to 65% and 45%, respectively, after 24 h, indicating that metals bound to metallothionein decreased more quickly than 35S-counts. These results suggest that metals were released from metallothionein and were excreted into the medium. However, 35S- and 109Cd-counts in Cd-thionein changed very little, if at all, in the cells even after a 24-h incubation period. Our data strongly suggest that Zn- and Cu-thionein are degraded in the cells, but that Cd-thionein remains longer than either Zn- or Cu-thionein. When cells containing Zn-thionein were incubated in metal-free medium, Zn-thionein was digested in the cells and peptide fragments ranging about 200-400 daltons were excreted from the cells.     

Induction of metallothionein-I mRNA in cultured cells by heavy metals and iodoacetate: evidence for gratuitous inducers.

A mouse hepatocyte cell line selected for growth in 80 microM CdSO4 (Cdr80 cells) was used to test the role of metallothioneins in heavy metal detoxification. The cadmium-resistant (Cdr80) cells have double minute chromosomes carrying amplified copies of the metallothionein-I gene and accumulate ca. 20-fold more metallothionein-I mRNA than unselected cadmium-sensitive (Cds) cells after optimal Cd stimulation. As a consequence, the amount of Cd which inhibits DNA synthesis by 50% is ca. 7.5-fold higher in Cdr80 cells than in Cds cells. Cds and Cdr80 cells were compared in terms of their resistance to other heavy metals. The results indicate that although Zn, Cu, Hg, Ag, Co, Ni, and Bi induce metallothionein-I mRNA accumulation in both Cdr80 and Cds cells, the Cdr80 cells show increased resistance to only a subset of these metals (Zn, Cu, Hg, and Bi). This suggests that not all metals which induce metallothionein mRNA are detoxified by metallothionein and argues against autoregulation of metallothionein genes. Metallothionein-I mRNA is also induced by iodoacetate, suggesting that the regulatory molecule has sensitive sulfhydryl groups.     

Properties of metallothionein induced by zinc, copper and cadmium in the frog, Xenopus laevis

1. Repeated injections of zinc (Zn) and copper (Cu) into the frog Xenopus laevis caused accumulations of the respective metals in the liver and kidney. 2. The accumulated metals in the liver supernatant fractions were present as Zn- and Cu-binding proteins of the same properties as that of metallothionein (MT) induced by cadmium (Cd) injections. 3. The affinity of Zn, Cu and Cd ions to the metal-binding protein was in the decreasing order of Cu, Cd and Zn. 4. The Xenopus MT induced by Cd was unstable and disrupted easily to give two peaks as if the MT consists of two isometallothioneins.     

Influence of cadmium on the distribution of the essential trace elements zinc and copper in the liver and kidneys of rats

Cd induced changes of Zn and Cd distribution in the liver and kidneys were studied in relation to Cd metallothionein (MT) synthesis. Wistar male rats were given CdCl₂ by sc injection of .8, 1.5, and 3.0 mg Cd/kg three times a week for three weeks. Cd levels of liver and kidneys increased with the increment of Cd dosage and 80–90% of Cd was found in the cytosol. The MT fractions contained 80–89% cytosolic Cd in the liver and 55–75% Cd in the kidneys. Zn concentrations in the liver increased following Cd administration, But Zn in the kidneys showed only slight increase. There was a distinct decrease of Cu concentration in the liver of the 3.0 mg group. In contrast, Cu concentrations in the kidneys increased about three times in the .8 and 1.5 mg Cd groups, but Cu in the 3.0 mg group showed only 1.5 times increase. The changes of these metal concentrations were observed mainly in the cytosol. Non-MT-Cd in the kidneys was maximum in the 1.5 mg group, but the 3.0 mg group showed significant decrease. In parallel with this decrease of Cd, Cu and Zn in the kidneys showed similar decrease. When the kidneys are injured, Zn and Cu appear to leak from this organ.     

Identification of metallothionein in parenchymal and non-parenchymal liver cells of the adult rat.

Parenchymal and non-parenchymal cells were isolated from the livers of control, starved, Zn2+-injected and Cd2+-injected rats. Parenchymal cells were prepared by differential centrifugation after perfusion of the liver with collagenase. Non-parenchymal cells were separated from parenchymal cells by unit-gravity sedimentation and differential centrifugation. Yields of 2 x 10(8) non-parenchymal cells with greater than 95% viability and less than 0.2% contamination with parenchymal cells were obtained without exposing cells to Pronase. Metallothioneins-I and -II were identified in parenchymal cells and non-parenchymal cells from Zn2+-treated rats. The metallothionein contents of parenchymal cells, non-parenchymal cells and intact liver were quantified by a competitive 203Hg-binding assay. Administration of heavy-metal salts significantly increased the metallothionein content of both cell populations, although the concentration of the protein was approx. 2.5-fold greater in parenchymal cells than in non-parenchymal cells. Overnight starvation increased the metallothionein content of parenchymal cells without altering that of non-parenchymal cells. The potential significance of this differential response by different liver cell types with regard to the influence of Zn2+ on stress-mediated alterations in hepatic metabolism is discussed.     

雌二醇、壬基酚、多氯联苯、镉和锌暴露对唐鱼体内超氧化物歧化酶活性的影响

目的 研究雌二醇(E2)、壬基酚(NP)、多氯联苯(PCBs)、镉(Cd2+)和锌(Zn2+)单独以及联合暴露对唐鱼体内SOD酶活力的影响.方法设计不同浓度的单一物质及混合物对唐鱼14 d暴露染毒,定量测定7、14d体内SOD酶活力的变化.结果 ①低浓度E2处理唐鱼7d可诱导SOD活性显著上升,时间延长、浓度升高时SOD活性无明显变化;②低浓度NP对SOD活性没有明显的影响,高浓度NP使SOD活性极显著上升;③中高浓度组PCBs处理7、14 d,低浓度组PCBs处理14 d时SOD活性被抑制,抑制程度随着时间的延长和浓度的升高有增强的趋势;④Cd2+、Zn2+各浓度组都对唐鱼体内的SOD活性产生了一定的抑制作用,并且抑制程度随着时间的延长而加深.结论 E2、NP、PCBs、Cd2+、Zn2+对唐鱼的SOD酶活力有明显影响,它们单独作用时的SOD活性与暴露浓度之间存在良好的剂量-效应关系.联合作用效应与暴露时间和(或)各物质浓度有关,大部分表现为毒性增强.     

嗜热四膜虫金属硫蛋白基因MMT2和MTT4的表达规律比较与进化模式分析

多个金属硫蛋白基因异构型已在四膜虫中被鉴定,这些异构型可分为7a和7b两个亚家族。该文利用实时荧光定量PCR技术检测了嗜热四膜虫金属硫蛋白基因MTT2和MTT4在Hg、Cu、Cd、Zn、H2O2暴露下的表达水平,结果显示两者表达规律相似,均为:Cu暴露下上调最高(>200倍),Hg次之,Cd、Zn上调倍数不大,H2O2有下调趋势。此表达规律明显有别于7a亚家族,具有7b亚家族的表达特征。同种诱导物暴露下MTT4的上调表达幅度比MTT2高出数倍,结合生物信息学分析结果,推测可能与MTT2和MTT4上游调控元件(如AP-1、MRE等)的数量差异有关。基于MTT2和MTT4在结构和功能上的高度相似性,推测两者可能是经近期基因复制事件产生,并遵循基因剂量模型进化而来。     

Effects of pretreatment with cadmium on the discriminative uptake of subsequent cadmium, copper or zinc by the liver

1. Effects of pretreatment with cadmium (Cd) on the uptake by the liver of subsequent Cd, copper (Cu) and zinc (Zn) were examined at two different time intervals to elucidate the biological discrimination mechanism among metals of similar chemical properties. 2. Pretreatment with 0.3 mg Cd/kg body wt 6 hr but not 24 hr before a subsequent dose of 0.8 mg metal/kg body wt enhanced the disappearance rate from plasma and accumulation rate in the liver of Cu (and Zn) but not of Cd. 3. Synthesis of metallothionein was induced with different time-courses depending on the time interval between the pretreatment and subsequent treatment, which coincided with the accumulation curves for Cu (and Zn) but not for Cd. 4. Although uptake of Cd was not enhanced by any pretreatment, metallothionein synthesis was enhanced depending on the timing of pretreatment.