The antiestrogen-like activity and reproductive toxicity of 2,6-DCBQ on female zebrafish upon sub-chronic exposure

2,6-Dichloro-1,4-benzoquinone (2,6-DCBQ), an emerging water disinfection by-product, is widely detected in water resources. However, its potential effects on the reproductive system are largely unknown. Here, we investigated the long-term effects of 2,6-DCBQ on gonadal development by exposing zebrafish from 15 to 180 days postfertilization (dpf). Following exposure to 2,6-DCBQ (20 and 100 µg/L), female-specific effects including delayed puberty onset, retarded ovarian growth and breakdown of the zona radiata were observed, resulting in subfertility in adult females. Adverse effects in folliculogenesis disappeared two months after cessation of 2,6-DCBQ administration. In contrast, no adverse impacts were noted in male testes. The effects on females were associated with significant reduction in 17β-estradiol (E2) level, suggesting a role for 2,6-DCBQ in anti-estrogenic activity. E2 level change in blood was further supported by dysregulated expression of genes (cyp19a1a, fshb, kiss3, esr2b, vtg1, and vtg3) related to the hypothalamic-pituitary-gonad-liver (HPGL) axis. The present study demonstrates for the first time that 2,6-DCBQ induces reproductive impairments in female zebrafish through disrupting 17β-estradiol level.     

Effect of triazophos,fipronil and their mixture on miRNA expression in adult zebrafish

MicroRNA (miRNA) plays a crucial role in gene expression regulation. However, no data are available on change of miRNA expression of zebrafish (Danio rerio) after treatment with pesticides. We evaluated the effect of fipronil (5-amino-1-[2, 6-dichloro-4-(trifluoromethyl) phenyl]-4-[(trifluoromethyl) sulfinyl]-1H-pyrazole-3-carbonitrile) and triazophos (3-(O, O-diethyl)-1-phenyl thiophosphoryl-1, 2, 4-triazol) and their mixture on miRNA expression in zebrafish. MiRNA expression profiles in zebrafish were altered after treatment with these chemicals. An association between these chemicals and the expression of 21 miRNAs was found 96 h after treatment. Among them, 14 miRNAs were differentially expressed due to the treatments with fipronil, triazophos and their mixture; 5 miRNAs showed altered expression level after treatment with formulations of these chemicals; miR-29b and miR-738 were differentially expressed after treatment with adjuvants. MiRNAs might present a novel toxicological response that could be used as a toxicological biomarker and have a different direction for future investigations of their association with miRNAs involved in chemical related diseases.     

Genotoxic effects of binary mixtures of xenoandrogens (tributyltin, triphenyltin) and a xenoestrogen (ethinylestradiol) in a partial life-cycle test with Zebrafish (Danio rerio)

A partial life-cycle test with the model fish Danio rerio was performed in order to evaluate the genotoxic potential of binary mixtures of xenoandrogenic (tributyltin--TBT; triphenyltin--TPT) and an estrogenic compound (ethinylestradiol--EE2). Five days post-fertilisation larvae were diet-exposed to environmental relevant concentrations of TBT and TPT (25 ng/g-100 ng/g), and water-exposed to ethinylestradiol (3.5 ng/L) for a four-month period; binary mixtures of TBT plus EE2 and TPT plus EE2 were run in parallel. The erythrocytic nuclear abnormalities (ENA) assay in circulating erythrocytes was used to evaluate genotoxicity in the end of the four-month exposure period. A significant increase (p<0.05, Kruskall-Wallis non-parametric ANOVA) in ENA frequency, in comparison with control animals, was observed in those animals exposed to TBT and TPT (the highest doses only), and to EE2 and the binary mixtures, although neither synergistic nor additive effects of the tested compounds were evident. Overall, the results clearly indicate that chronic exposure to low levels of TBT, TPT, EE2 and binary mixtures of TBT plus EE2 and TPT plus EE2 are genotoxic to zebrafish, which may suggest that wild fish populations may be under increased DNA damage in areas contaminated by these endocrine disrupting chemicals.     

石棉表面改性对石棉所致人胚肺细胞内游离钙离子浓度改变的抑制作用

为了探讨研究温石棉及用柠檬酸铝、混合稀土或亚硒酸钠预处理的温石棉对人胚肺（HEL）细胞内游离钙离子（[Ca^2＋]i）浓度的影响。用Flou3／AM标记体外培养的HEL细胞内Ca^2＋后，将不同浓度的温石棉及用柠檬酸铝、混合稀土或亚硒酸钠溶液浸泡过的温石棉与HEL细胞作用1h，荧光分光光度计测定细胞内[Ca^2＋]i。结果显示，20、40、80μg／ml温石棉单独作用于HEL细胞1h，可使HEL细胞内[Ca^2＋]i浓度分别升高3．0％、53．1％和116．7％，且具有明显的剂量-效应关系（r=0．976，P=0．024）。经三种化合物预处理的温石棉组HEL细胞内[Ca^2＋]i浓度均明显低于未处理温石棉组。得出结论，温石棉可致人胚肺细胞内游离钙离子浓度升高，但用三种化合物预处理温石棉，均可抑制温石棉所致人胚肺细胞内游离钙离子浓度的升高。     

联苯胺类化合物对斑马鱼胚胎发育毒性的初步研究

应用斑马鱼胚胎发育技术,对4种联苯胺类化合物(联苯胺、3,3'-二甲氧基联苯胺、3,3'-二甲基联苯胺、3,3'-二氯联苯胺)的毒性进行了测定.结果表明,这4种化合物对斑马鱼胚胎发育均有明显抑制作用,可以造成仔鱼畸形甚至死亡,具有特定的最敏感毒理学终点及作用时间;联苯胺苯环上的取代基可以增强联苯胺的亲电作用或亲核作用,从而增强其胚胎毒性,其中尤以卤代产物(3,3'-二氯联苯胺)最为显著.其毒性排列顺序大致为:3,3'-二氯联苯胺>3,3'-二甲基联苯胺>3,3'-二甲氧基联苯胺>联苯胺;4种联苯胺的斑马鱼胚胎毒性数据与lgKow没有显著相关性,推测这4种联苯胺的毒性机理属体内反应型,毒物参与生理代谢.     

Cypermethrin has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish (Danio rerio)

Cypermethrin (CYP), a widely used Type II pyrethroid pesticide, is one of the most common contaminants in the freshwater aquatic system. We studied the effects of CYP exposure on the induction of hepatic oxidative stress, DNA damage and the alteration of gene expression related to apoptosis in adult zebrafish. Hepatic mRNA levels for the genes encoding antioxidant proteins, such as Cu/Zn-Sod, Mn-Sod, Cat, and Gpx, were significantly upregulated when zebrafish were exposed to various concentrations of CYP for 4 or 8 days. In addition, the main genes related to fatty acid β-oxidation and the mitochondrial genes related to respiration and ATP synthesis were also significantly upregulated after exposure to high concentrations (1 and 3 μg L⁻¹) of CYP for 4 or 8 days. Moreover, in a comet assay of zebrafish hepatocytes, tail DNA, tail length, tail moment and Olive tail moment increased in a concentration-dependent manner. The significant induction (p < 0.01) of all four parameters observed with CYP concentrations of 0.3 μg L⁻¹ or higher suggests that heavy DNA damage was induced even at low levels. Furthermore, several apoptosis- related genes, such as p53, Apaf1 and Cas3, were significantly upregulated after CYP exposure, and Bcl2/Bax expression ratio decreased, especially in groups treated with 1 and 3 μg L⁻¹ CYP for 8 days. Taken together, our results suggested that CYP has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in zebrafish. This information will be helpful in fully understanding the mechanism of aquatic toxicology induced by CYP in fish.     

Cd2、Hg2+、Cr6+和pb2对黑点青鳉(Oryzias melastigma)早期生活阶段的毒性效应研究

近岸海域重金属污染不容乐观,作为“一种潜在的海洋模式鱼种”黑点青鳉(marine medaka,Oryzias maelastigma)对不同重金属胁迫的响应水平却鲜有报道.为探究并比较海水鱼种黑点青鳉早期生活阶段对Cd2+、Hg2+、Cr6 +和pb2 +等不同重金属胁迫的响应,在实验室通过半静态方式,对黑点青鳉受精...     

纳米ZnO对斑马鱼胚胎抗氧化酶系统的影响

为评价纳米ZnO的水生态毒性,以斑马鱼胚胎为受试生物,研究了纳米ZnO对其抗氧化酶系统的影响.结果发现,纳米ZnO能够降低斑马鱼胚胎谷胱甘肽(GSH)含量,抑制过氧化氢酶(CAT)和超氧化物歧化酶(SOD)活性,引起胚胎脂质过氧化水平增大,证明氧化应激是纳米ZnO抑制斑马鱼胚胎孵化的作用机制之一.     

2,3,7,8-四氯苯并二噁英（TCDD）短期染毒可造成着床前胚胎丢失并伴随雌性生殖器官中毒物相关蛋白的诱导表达（Short Time Exposure of TCDD Causing Loss of Implantation Failure of Embryos and Its Associated with Induced Expression of Relevant Proteins）

为探明妊娠早期胚胎的丢失是否与卵巢、输卵管、子宫组织受到2,3,7,8-四氯苯并二噁英(TCDD)直接毒害有关,检测了NIH小鼠胚胎着床前期和后期TCDD暴露对胚胎毒性影响的敏感性,并利用免疫组化方法分析了模型动物肝脏、子宫、输卵管和卵巢组织中TCDD所引起的AhR、ARNT以及Cyp1a2分子标记物的变化.检测发现:妊娠第9d,100ng·kg-1·d-1剂量TCDD经口染毒,造成胚胎着床数量减少,且着床前期暴露的影响大于着床后期;子宫蜕膜反应受到明显抑制;胚胎迁移率没有明显变化,但胚胎数量减少.免疫组织化学分析发现正常组小鼠的肝脏、子宫、输卵管和卵巢组织中有AhR和Cyp1a2弱阳性信号表达,ARNT有细胞核的强阳性信号表达;妊娠第1～8d、第1～3d和第4～8d处理组小鼠肝脏、子宫、输卵管和卵巢组织中的AhR、Cyp1a2的阳性面积和光密度值均高于正常组;随处理时间和组织蓄积量的增加,ARNT在组织中的变化由胞核(妊娠第1～3d组)表达到胞浆(妊娠第4～8d组)表达,然后完全无表达(妊娠第1～8d组).以上研究结果表明:TCDD对早期妊娠小鼠子宫、输卵管和卵巢组织中的AhR、ARNT和Cyp1a2的激活和代谢方式与肝脏相同,说明雌性生殖系统中的组织有TCDD蓄积和代谢活性,这可能是导致早期胚胎迁移、着床等过程改变,造成胚胎丢失的重要原因.     

热带爪蟾(Xenopus tropicalis)胚胎在生态毒理学中的应用前景

首先介绍了热带爪蟾(Xenopus tropicalis)作为新的模式动物的发展过程以及热带爪蟾基因组学方面的研究进展,概述了非洲爪蟾(X.laevis)胚胎在生态毒理学中的应用。在此基础上,详细讨论了热带爪蟾胚胎在污染物致毒效应和作用机理、毒理学理论和环境样品生态毒性检测等方面的应用现状和前景。最后,简要介绍了目前国内外有关热带爪蟾网站、书籍和养殖等方面的信息资源。