4-n-Nonylphenol and 17-β estradiol may induce common DNA effects in developing barnacle larvae

There is a growing concern over the potential effects of environmental endocrine disrupters on both human and wildlife populations. However, to date, minimal research has been conducted to determine the effect of estrogens and xenoestrogens at the DNA level. In this study, we used the random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of barnacle larvae that had been exposed to 17β-estradiol (E₂) and low concentrations of 4-n-nonylphenol (NP). DNA effects include DNA damage as well as mutations and possibly other effects at the DNA level that can be induced by chemical or physical agents that directly and/or indirectly interact with genomic DNA. Not only did exposure to NP and E₂ induce changes in RAPD profiles in the exposed barnacle larvae when compared to control patterns, but also, and more importantly, there were similarities in the RAPD modifications in the exposed populations that had been treated to either chemical. We propose that NP and E₂ induced some common DNA effects in barnacle larvae and that these specific modifications in RAPD patterns may arise as a consequence of hot spot DNA damage (e.g. DNA adducts) and/or mutations (point mutations or genomic rearrangements). This could help to explain how xenoestrogens mimic the effects produced by natural estrogens. In conclusion, in the field of endocrine disruption, the study of DNA effects induced by estrogens and/or xenoestrogens warrants further investigation. Indeed, changes at the DNA level may be the precursors of some of the numerous effects reported at higher levels of biological organisation such as the feminization of males, developmental abnormalities, and infertility.     

长江、汉江水源水及其自来水中有机物生物毒性的比较

应用XAD-2大孔树脂对长江、汉江(武汉)水源水及其自来水水样中的有机物进行吸附和富集,经洗脱、浓缩、干燥,提取其非挥发性有机物(NVOCs),通过致突变试验(Ames test)和重组酵母雌激素测评试验(YES)分别检测其致突变性与类雌激素效应.结果显示,2008年平水期,水中NVOCs除了长江出厂水对TA98(＋S9)的致突变性检测结果为阴性外,其他水样均为阳性;丰水期除长江水源水外,其他水样对TA98的致突变性为阳性;枯水期在不加S9的条件下,仅长江出厂水、管网末梢水对TA98的致突变性为阳性,其余水样均为阴性.各水样的NVOCs对TA100(±S9)的结果均为阴性.平水期长江、汉江水源水和枯水期长江水源水中的NVOCs均可检出类雌激素效应,丰水期水源水以及各水文期自来水中的NVOCs未显类雌激素效应.2007～2009年不同水文期长江和汉江自来水中NVOCs的致突变比活性均高于水源水;长江、汉江各水样中具有致突变作用的有机物主要为移码型直接致突变物.长江水源水NVOCs的类雌激素效应高于汉江,经自来水厂处理后的出厂水以及管网末梢水中的NVOCs未检出类雌激素效应.     

多溴二苯醚类化合物干扰人雌激素受体alpha活性机理的理论研究

基于分子对接方法探讨了多溴二苯醚(PBDEs)类化合物与人雌激素受体α亚型间的分子作用机理.对多溴二苯醚类化合物是否具有拟雌激素功能的研究得出:可通过对接打分值和化合物结构特征来推测PBDEs母体化合物是否具有拟雌激素活性;对HO-PBDEs,与氨基酸残基GLU53和/或ARG394形成氢键可能是影响其拟雌激素活性的重要因素;对MeO-PBDEs,疏水MeO-位于结合腔的疏水中部有利于拟雌激素活性.从结构及构象分析得出,邻位疏水基(Br-、MeO-)有利于PBDEs类化合物的拟雌激素活性.同时对多溴二苯醚类化合物是否具有抗雌激素功能的结合特征研究发现,表现出抗雌激素活性的部分PBDEs类化合物伸进通常被雌激素受体拮抗剂雷洛昔芬和4-羟基它莫西芬的烷基胺侧链占据的通道,而大多数未表现出抗雌激素活性的PBDEs类化合物的结合模式类似雌激素受体激动剂17β-雌二醇,位于结合腔,没有伸进通道.本研究从化合物结构及化合物在受体内结合的构象特征上解释化合物活性不同的原因,以期能够利用构象分析得到的结果进行筛选.     

Interactions of Endocrine-active environmental chemicals with the nuclear receptor PXR

There is growing concern about the human-health impact of environmental chemicals that have the potential to disrupt normal endocrine function. Endocrine disrupting chemicals (EDCs) include structurally diverse organochlorine pesticides, polychlorinated biphenyls (PCBs), plasticizers, fungicides, herbicides and pharmaceutical compounds, and can have a profound impact on development, and on reproductive, neurological and immune system functions. While many studies have focused on the role of androgen receptor, estrogen receptor and aryl hydrocarbon receptor in mediating the effects of EDCs, other nuclear receptors that regulate steroid hormone action and metabolism may also serve as targets of EDC action. This review focuses on two classes of EDCs, PCBs and phthalate monoesters, both of which have been shown to interact with pregnane X receptor (PXR), a member of the nuclear receptor superfamily that regulates a large number of target genes, many of which have important roles in steroid metabolism and transport. Recent findings on the ability of PCBs and phthalate monoesters to activate PXR are discussed and the potential role of PXR and other intracellular receptor proteins in mediating toxicities associated with EDC exposure is considered. Finally, we discuss several gaps in our knowledge regarding the actions of EDCs and the difficulties associated with the evaluation of risks associated with exposure to these endocrine active environmental chemicals.     

有机紫外防晒剂内分泌干扰效应研究进展

有机紫外防晒剂是一类新兴环境污染物,大量研究表明,部分有机紫外防晒剂能模拟生物体内多种激素的作用,干扰动物和人体正常的内分泌功能,造成严重的健康威胁.本文在大量文献调研的基础上,从个体水平、器官水平、细胞水平和分子水平4个层次,就有机紫外防晒剂对生殖系统、青春期发育、成活率、pS2蛋白、卵黄蛋白原及激素受体等多方面的影响进行了综述,以加深对紫外防晒剂的认识,也为紫外防晒剂的正确使用及相关部门对其进行科学管理提供参考.     

己烯雌酚降解菌固定化条件优化及其降解性能

以海藻酸钠为包埋剂,制备了己烯雌酚(DES)降解菌株S(Serratia sp.)的固定化菌剂,通过正交试验确定了菌株S的最佳固定化条件:海藻酸钠质量分数为4%,菌悬液与海藻酸钠重量比为1:2,CaCl₂质量分数为4%,交联时间为6h.菌株S的固定化菌剂比游离菌株更有优势;7d时,固定化菌株S对无机盐培养基中DES的去除率为83.84%,较游离菌株高22.84%.优化了固定化菌株S对溶液中DES的处理条件:固定化菌株S接入量为300g/L,初始底物浓度为20~40mg/L.固定化菌株S对DES浓度为40.01,37.90 ,33.52μg/L 3个排污口污水中DES的去除率分别达90.0%,96.0%,96.7%.利用固定化菌株处理实际污水中DES等雌激素具有应用潜力.     

邻苯二甲酸二(2-乙基己)酯(DEHP)和邻苯二甲酸单乙基己基酯(MEHP)长期暴露对海洋青鳉(Oryzias melastigma)内分泌干扰效应的评价

为探讨邻苯二甲酸二(2-乙基己)酯(DEHP)和邻苯二甲酸单乙基己基酯(MEHP)长期暴露对海水生物的内分泌干扰效应及机制,将孵化后1周的海洋青鳉(Oryzias melastigma)分别暴露于DEHP(0.1 mg·L-1和0.5 mg·L-1)和MEHP(0.1 mg·L-1和0.5 mg·L-1)6个月。结果显示:DEHP显著增加了雌性和雄性海洋青鳉的肝指数,而MEHP只在高剂量时显著增加雄性青鳉的肝指数。对于雌性青鳉,DEHP暴露后肝脏雌激素相关基因ERα、ERβ、ERγ、VTG1、VTG2、ChgH和ChgL的表达水平显著上调,而对于雄性青鳉,DEHP暴露后,只有肝脏ERβ的表达水平显著上调。相比之下,MEHP暴露对雌性和雄性青鳉肝VTG和Chg基因表达无显著影响。DEHP激活了雌性和雄性青鳉的肝过氧化物增殖激活受体PPARα和PPARγ,而MEHP只在低剂量时上调了雄性青鳉PPARγ的表达。在雌性和雄性青鳉体内,VTG和Chg的表达与ERα和ERγ的表达显著相关,并且ER与PPAR也显著相关。研究表明,DEHP长期暴露可通过激活肝性激素受体调控肝雌激素响应基因(VTG和Chg)和过氧化物增殖激活受体(PPARα和PPARγ)的表达而对海洋青鳉产生内分泌干扰效应,并且显示出性别特异性。MEHP对海洋青鳉的内分泌干扰效应弱于DEHP。     

金雀异黄素对雄性斑马鱼卵黄原蛋白mRNA的诱导机理

采用质量浓度分别为1、10、100、1 000 μg/L的金雀异黄素对成年雄性斑马鱼进行水体暴露28 d,通过real-time PCR技术检测雄性斑马鱼肝脏内Vtg-1(卵黄原蛋白)、ERα和ERβ1 mRNA(雌激素受体)的表达水平,并利用同位素放射免疫法检测雄性斑马鱼性腺w(睾酮)和w(E₂)(E₂为17β-estradiol,17β-雌二醇). 结果表明:10~1 000 μg/L金雀异黄素显著诱导了雄性斑马鱼肝脏Vtg-1 mRNA的表达,二者呈剂量-效应关系,同时显著上调了雌激素受体ERβ1 mRNA的表达;但1~1 000 μg/L金雀异黄素却极显著地抑制了雄性斑马鱼肝脏内ERα mRNA的表达,同时造成雄性斑马鱼性腺w(E₂)的升高和w(睾酮)的降低. 研究结果显示,金雀异黄素可能通过提高雄性斑马鱼体内ERβ1mRNA的表达和内源w(E₂),增强ERβ1对内源雌激素的介导作用,从而促进Vtg-1的表达.      

腺苷酸环化酶抑制剂体外筛选模型的建立及应用

腺苷酸环化酶(adenylate cyclase,AC)通过调节环磷酸腺苷(cyclic adenosine cyclophosphate,cAMP)的合成从而调控细胞相关功能.在卵巢中,cAMP调控卵母细胞的发育及卵子的成熟,cAMP含量变化影响下游芳香化酶的表达,进而影响雌激素的生物合成.为开发靶向AC的候选药物,根据AC1、AC2两个活性区域,利用大肠杆菌系统克隆表达可溶ⅠC1-ⅡC2重组融合蛋白,使用ATP检测试剂盒,检测ⅠC1-ⅡC2的催化活性,从而建立有效的ⅠC1-ⅡC2蛋白体外筛选抑制剂模型.利用该模型筛选了由2 861个化合物组成的文库.化合物来那替尼(neratinib)可明显抑制ⅠC1-ⅡC2蛋白活性,且呈浓度依赖性,其IC50为10.2±0.152 2μmol/L.进一步研究发现,来那替尼可显著降低人卵巢颗粒细胞KGN细胞中芳香化酶的表达,并且抑制雌激素的生物合成.本研究通过建立蛋白体外筛选模型发现来那替尼具有抑制AC的活性,结果可为开发新的靶向芳香化酶治疗多囊卵巢综合征等疾病的候选药物提供参考.(图5表1参28)     

重组基因酵母法研究酞酸酯类化合物的类雌激素作用

为了检测五种酞酸酯类化合物的类雌激素活性,采用重组人雌激受体α、相关效应元件以及β-半乳糖苷酶报告基因酵母分别测定17β-雌二醇(E)2、邻苯二甲酸二甲酯(DMP)、邻苯二甲酸二乙酯(DEP)、邻苯二甲酸二丁酯(DBP)、邻苯二甲酸二辛酯(DOP)、邻苯二甲酸丁苄酯(BBP)单独作用时诱导酶活并计算其EC50。通过各化合物与E2的EC50的比以及最大β-半乳糖苷酶活性来比较酞酸酯类化合物的类雌激素活性。五种酞酸酯类化合物的实验浓度范围为1.0×10-3~1.0×10-11 mol/L。结果表明,DEP(1×10-7~1×10-9 mol/L)、DBP(1×10-6~1×10-9 mol/L)、BBP(1×10-3~1×10-6 mol/L)具有明显的类雌激素活性,诱导的最大酶活分别为:1.515、0.832 2、1.669,而DMP和DOP并未检出明显的类雌激素活性。DEP、DBP、BBP与E2的EC50的比分别为8.85×10-2、2.54×10-2、8.82×10-6。说明DEP、DBP在低浓度就表现出类雌激素活性,而BBP在较高浓度才表现出类雌激素活性。