褪黑素对邻苯二甲酸(2-乙基己基)酯致雄性小鼠生殖细胞DNA损伤的拮抗作用

目的探讨邻苯二甲酸(2-乙基己基)酯(DEHP)致小鼠睾丸细胞DNA损伤及褪黑素(MT)对此损伤的拮抗作用。方法将40只CL57BL/6J雄性小鼠随机分为4组,包括对照组、MT组、DEHP组和MT+DEHP联合组。MT采用腹腔注射(剂量为15 mg·kg-1),DEHP灌胃染毒(染毒剂量为1000 mg·kg-1),每天染毒1次,连续30 d。检测睾丸组织中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活力和丙二醛(MDA)、8-羟基脱氧鸟嘌呤(8-OHd G)含量。单细胞凝胶电泳(彗星实验)检测睾丸细胞DNA损伤,慧星图像软件测定慧星尾长、慧尾DNA百分含量、尾矩及Olive尾矩。结果与对照组比较,DEHP组小鼠睾丸组织GSH-Px和SOD活力降低,MDA和8-OHd G含量增加,睾丸细胞彗星尾长、彗尾DNA百分含量、尾矩、Olive尾矩均显著增加,差异均有统计学意义(P<0.05);与DEHP组比较,MT+DEHP联合组小鼠睾丸组织GSH-Px和SOD活力升高,MDA和8-OHd G含量降低,睾丸细胞DNA损伤程度减轻,差异均有统计学意义(P<0.05)。结论 DEHP造成小鼠睾丸明显的氧化应激,并引起睾丸细胞DNA的损伤;MT可拮抗因DEHP染毒导致的睾丸氧化损伤。     

邻苯二甲酸二(2-乙基己基)酯长期暴露对鲤鱼毒理学指标的影响

以吐温80和水为对照组,用5 mg·L-1、20 mg·L-1、80 mg·L-1、160 mg·L-1的邻苯二甲酸二(2-乙基己基)酯(DEHP)对鲤鱼暴露染毒20 d,用试剂盒等方法研究了DEHP对鲤鱼肝脏抗氧化防御系统、肝解毒酶、能量代谢酶和肝损伤标志酶的影响。结果表明,与水空白对照组和吐温80组相比,各浓度DEHP组,肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力和抗超氧阴离子自由基能力的差异均无统计学意义,谷胱甘肽过氧化物酶(GSHPX)活力表现为低促高抑,160 mg·L-1组抗羟自由基能力显著降低,丙二醛(MDA)含量显著升高。研究结果还表明,在DEHP实验浓度范围内,谷胱甘肽硫转移酶、葡萄糖-6-磷酸脱氢酶活力表现为低促高抑,而160 mg·L-1组7-乙氧基-3-异吩噁唑酮-脱乙基酶、Na+K+-ATPase、单胺氧化酶(MAO)活力和各DEHP组Ca2+Mg2+-ATPase活力均受到了显著抑制,20 mg·L-1以上组肝γ-谷氨酰转肽酶活力则显著升高。结果显示,一定浓度的DEHP能损伤肝脏组织,并对肝组织的毒理学指标产生影响。     

邻苯二甲酸二（2-乙基己基）酯对睾丸的毒性作用及机制

邻苯二甲酸二（2-乙基己基）酯（DEHP）是目前使用最广泛的增塑剂之一,DEHP具有毒性,长期暴露会对机体的多个系统产生损害,特别是对雄性生殖系统的毒性作用更为明显。DEHP通过诱导氧化应激、调控细胞自噬,促进生精细胞和睾丸间质细胞凋亡、抑制睾酮合成、破坏血-睾屏障、诱导睾丸支持细胞铁死亡以及影响子代雄性的表观遗传等,造成生殖器官的病理损伤。本文就DEHP对睾丸的毒性作用及机制进行综述,拟为男性生殖障碍的防治研究提供新思路。     

邻苯二甲酸二乙基己酯长期暴露对鲤鱼粘液细胞的影响

利用阿利新蓝和过碘酸雪夫氏(AB-PAS)染色方法,研究了邻苯二甲酸二乙基己酯(DEHP)对鲤鱼鳃及消化道粘液细胞密度的影响,实验分为空白对照组、吐温80组及5 mg/L、20 mg/L、80 mg/L、160 mg/L DEHP组,染毒20 d。结果表明,吐温80组与空白对照组相比差异不明显。除5 mg/L组口腔上皮处粘液细胞密度显著低于吐温80组外,在实验剂量范围内,鳃丝、口腔上皮、前肠、后肠粘液细胞密度均极显著低于吐温80组。中肠处粘液细胞密度5 mg/L、20 mg/L组与吐温80组相比差异不显著,80 mg/L组显著低于吐温80组,160 mg/L组极显著低于吐温80组。鳃丝、口腔上皮、中肠处粘液细胞密度均随DEHP浓度的增大而减小,而前肠和后肠在80 mg/L组粘液细胞密度最小,降幅最大。实验结果还表明:在实验剂量范围内,与吐温80组相比,DEHP能降低鳃丝、口腔上皮处Ⅱ、Ⅳ型,前肠Ⅰ、Ⅱ、Ⅳ型,中肠Ⅰ、Ⅲ、Ⅳ型,后肠Ⅱ、Ⅲ型粘液细胞密度。DEHP能提高鳃丝Ⅰ、Ⅲ型,口腔上皮和前肠Ⅲ型,中肠Ⅱ型和后肠Ⅰ型粘液细胞密度。DEHP能减小鳃丝及消化道粘液细胞的密度,对各类型粘液细胞密度的影响因部位不同、DEHP浓度不同而异。     

邻苯二甲酸二乙基己酯对翡翠贻贝(Perna viridis)生化指标的影响

实验条件下,研究邻苯二甲酸二乙基己酯(DEHP)5个浓度组(0、0.38、1.92、9.60和48.00mg.L-1)长时间胁迫下翡翠贻贝(Perna viridis)内脏团和外套膜中抗氧化酶(SOD和CAT)活性和丙二醛(MDA)含量的变化,以及胁迫解除后这些指标的恢复情况。结果表明:在胁迫过程中,翡翠贻贝内脏团SOD活性表现为先显著升高,随后受抑制而逐渐降低(P<0.05),CAT活性则表现为先被抑制后受诱导,15d后恢复到对照组水平,MDA含量呈显著增加的趋势(P<0.05);外套膜中的SOD活性在胁迫初期在低浓度组被抑制,而在高浓度组则被诱导(P<0.05),4d后SOD活性逐渐恢复到对照组水平,各浓度组MDA含量均出现明显的增加(P<0.05);净化阶段,低浓度组(0.38mg.L-1)内脏团SOD活性和CAT活性逐渐恢复到对照组水平,但MDA含量升高;净化7d后,除高浓度组(48.00mg.L-1)外,其余浓度组外套膜中SOD活性均已经恢复到对照组水平,MDA含量也没有出现明显升高的现象。研究表明,DEHP对翡翠贻贝内脏团和外套膜抗氧化防御系统酶具有明显的影响,DEHP诱导引起2种组织内脂质过氧化损伤,并且短期内这种损伤无法消除。     

邻苯二甲酸二乙基己酯对鲤非特异性免疫的影响及遗传毒性

以水和吐温-80为对照,用5、20、80、160 mg/L的邻苯二甲酸二乙基己酯（DEHP）对鲤染毒20d,研究了DEHP对鲤非特异性免疫功能的影响及遗传毒性。结果表明: 吐温-80组与水对照组相比除红细胞总核异常率显著升高外,所测定的各项指标差异均不显著。80、160 mg/L组白细胞吞噬活力、吞噬指数,血清抗菌活力,溶菌酶活性均显著低于水对照组和吐温-80组（P0.05或P0.01）,且20 mg/L组白细胞吞噬活力、吞噬指数显著低于水对照组。与吐温-80组相比,160 mg/L组血清C3含量显著降低,5、20、80 mg/L组C3含量,5、20、80、160 mg/L组C4含量则无显著性差异。20、80、160 mg/L组C3含量,160 mg/L组C4含量显著低于水对照组（P0.05或P0.01）。160 mg/L组红细胞微核率显著高于吐温-80组,80、160 mg/L组红细胞微核率显著高于水对照组。在DEHP实验浓度范围内,红细胞核异常率、总核异常率,肝细胞DPC系数均显著高于水对照组和吐温-80组。一定浓度的DEHP对鲤具有免疫毒性和遗传毒性。       

邻苯二甲酸二(2-乙基己基)酯(DEHP)的细菌生物降解

邻苯二甲酸二(2-乙基己基)酯(dis(2-ethylhexyl)phthalate, DEHP)是一种人工合成的有机化合物,其作为增塑剂,因具有优良的柔韧性、可塑性和耐久性等特征而被广泛应用在个人护理、医疗器械、食品包装和塑料制造等领域。DEHP在土壤、水体和大气等环境中普遍存在,且由于DEHP是一种常见的内分泌干扰物,具有生殖毒性、免疫毒性以及神经毒性等,不仅给生态环境造成了威胁,而且可通过食物链进入人体给健康带来危害。因此,去除环境中的DEHP备受关注。DEHP的生物降解被认为是最绿色环保的降解方式,目前已有不少关于DEHP生物降解的研究报道。本文综述了DEHP的污染及危害、细菌生物降解现状,重点介绍了DEHP经酯键水解、β氧化、原儿茶酸降解、苯甲酸降解以及三羧酸循环等过程实现完全降解的途径,以及从基因层面阐述DEHP降解的分子机制,并针对DEHP生物降解目前还存在的问题进行总结和展望,为环境中DEHP有效生物降解提供参考。     

增塑剂邻苯二甲酸二(2-乙基己基)酯(DEHP)对绒毛外滋养层细胞侵袭和迁移的影响

为了探讨邻苯二甲酸二(2-乙基己基)酯(DEHP)对人绒毛外滋养层细胞HTR-8/Svneo侵袭和迁移的影响及机制,分别采用MTT法和流式细胞术确定DEHP的作用浓度,Transwell小室检测DEHP对细胞侵袭和迁移能力的影响,Rea;-time PCR和Western blot检测侵袭与迁移相关因子表达的变化。研究结果显示:100μmol/L及以上浓度的DEHP可促进HTR-8/Svneo细胞侵袭、迁移及MMP-2、MMP-9的表达,并上调p38、JNK的磷酸化,而DEHP的作用受p38与JNK的抑制剂拮抗。由此证明DEHP可通过p38、JNK信号通路上调MMP-2、MMP-9的表达,进而促进HTR-8/Svneo细胞侵袭和迁移。     

一株高效DEHP降解菌的分离、鉴定及其降解特性

【目的】分离得到高效的邻苯二甲酸二乙基己基酯(DEHP)降解菌。【方法】采用富集培养法筛选分离菌株,并对菌株进行驯化;通过PCR扩增得到其16S rRNA和gyrB基因序列,进行同源序列分析及分子系统发育树的构建,同时结合形态学观察和生理生化实验对菌株进行初步鉴定;采用高效液相色谱(HPLC)分析菌株对DEHP的降解特性。【结果】分离得到一株能以DEHP为唯一碳源和能源生长的菌株,命名为HS-NH1,初步鉴定其为戈登氏菌(Gordoniasp.)。菌株HS-NH1最适的生长和降解条件为30°C、pH 7.0,在此条件下,该菌株60 h内能够将浓度为500 mg/L的DEHP降解90%以上。高效液相色谱(HPLC)分析表明,菌株HS-NH1在降解DEHP过程中产生了一种重要的中间代谢产物——邻苯二甲酸。底物广谱性试验证明,菌株HS-NH1能够有效地利用多种常见的邻苯二甲酸酯(PAEs)与芳香族衍生物。【结论】筛选得到了一株DEHP降解菌Gordonia sp.HS-NH1,该菌降解效率高,具有良好的底物广谱性,在邻苯二甲酸酯类化合物的污染治理中将会有一定的应用潜力。     

DEHP对中国林蛙精巢中StAR、CYP17和CYP19基因表达的影响

为探讨邻苯二甲酸二(2-乙基己基)酯(di-2-ethylhexyl phthalate,DEHP)对两栖动物精巢类固醇激素合成的影响,将中国林蛙(Rana chensinensis)雄性成体分别暴露于浓度为10-7、10-6、10-5、10-4mol·L-1DEHP的水体,分别在暴露20、30和40d取其精巢,提取精巢总RNA,逆转录合成cDNA,通过荧光实时定量PCR检测StAR、CYP17和CYP19mRNA表达相对值。结果表明:与对照组相比,DEHP处理组StAR和CYP19基因表达均上调,CYP17基因表达下调;比较不同DEHP浓度和不同暴露时间对StAR、CYP17和CYP19mRNA表达相对值的影响,显示DEHP浓度变化对3个基因表达影响的规律性不强,而DEHP暴露时间的累积效应较明显;提示DEHP可通过干扰中国林蛙精巢中StAR、CYP17和CYP19基因表达,影响其相应关键酶的表达,从而干扰类固醇激素的合成,产生雌激素效应。     

High-performance liquid chromatographic method for the determination of di(2-ethylhexyl) phthalate in total parenteral nutrition and in plasma

A simple, rapid and sensitive reversed-phase high-performance liquid chromatographic method with UV detection was developed for the quantification of di(2-ethylhexyl) phthalate (DEHP) in parenteral nutrition admixtures containing fat emulsion and in plasma samples of children daily treated by total parenteral nutrition. The analyte and the internal standard, di-n-heptyl phthalate, were extracted twice using hexane and the organic layer separated and dried under nitrogen. The residues were reconstituted with acetonitrile and 20 μl was injected into a Waters Spherisorb C₁₈ column, the UV detector was set at 202 nm. The mobile phase was acetonitrile–aqueous buffer (triethylamine 0.08% adjusted to pH 2.8 with 1 M phosphoric acid) mixture (88:12, v/v) and it was pumped at 1 ml/min. Average recoveries were 97% or greater. This method was successfully used to investigate the amounts of DEHP which can leach from bags and tubing into fat emulsion and which could contaminate children under long-term parenteral nutrition. On the other hand, the circulating DEHP concentrations were estimated in four children under regular long-term parenteral nutrition.     

Biosorption of di(2-ethylhexyl)phthalate by seaweed biomass

Samples of various Sargassum species were collected in the Hong Kong marine environment and used for studies on biosorption of di(2-ethylhexyl)phthalate (DEHP). Batch adsorption experiments were carried out to determine the removal capacity and removal efficiency of the biosorbents. The DEHP removal ability was similar among beached seaweed and three freshly collected Sargassum species. Different physico-chemical factors were evaluated in order to enhance the performance of the biosorbents. Under optimized conditions (25 mg biomass, initial pH 4, 25 °C, 40 mg L^–1 DEHP), the mean removal capacity of beached seaweed and Sargassum siliquastrum was 5.68 and 6.54 mg g^–1, respectively. Examination of the Langmuir and Freundlich adsorption isotherms showed that the biosorption phenomenon by these biosorbents could well be described by these models. Desorption of DEHP was also assessed with methanol, which showed the most satisfactory desorbing ability. Further study in multiple adsorption–desorption of DEHP by the biosorbents demonstrated the reusability of both beached seaweed and S. siliquastrum for biosorption of DEHP.     

The effects of di(2-ethylhexyl) phthalate and/or selenium on trace element levels in different organs of rats

Di(2-ethylhexyl)phthalate (DEHP), a widely used plasticizer for synthetic polymers, is known to have endocrine disruptive potential, reproductive toxicity, and induces hepatic carcinogenesis in rodents. Selenium (Se) is a component of several selenoenzymes which are essential for cellular antioxidant defense and for the functions of mammalian reproductive system. The present study was designed to investigate the effects of DEHP exposure on trace element distribution in liver, testis, and kidney tissues and plasma of Se-deficient and Se-supplemented rats. Se deficiency was produced by feeding 3-week old Sprague-Dawley rats with ≤0.05 mg Se/kg diet for 5 weeks, and supplementation group were on 1 mg Se/kg diet. DEHP treated groups received 1000 mg/kg dose by gavage during the last 10 days of feeding period. Se, zinc (Zn), copper (Cu), iron (Fe) and manganese (Mn) levels were measured by inductively coupled plasma mass spectrometry (ICP-MS). Se supplementation caused significant increases in hepatic, renal, and testicular Se levels. With DEHP exposure, plasma Se and Zn, kidney Se, Cu and Mn levels were significantly decreased. Besides, liver Fe decreased markedly in all the DEHP-treated groups. Liver and kidney Mn levels decreased significantly in DEHP/SeD group compared to both DEHP and SeD groups. These results showed the potential of DEHP exposure and/or different Se status to modify the distribution pattern of essential trace elements in various tissues, the importance of which needs to be further evaluated.     

邻苯二甲酸酯在不同品种通菜-土壤系统中的累积效应研究

在邻苯二甲酸(2-乙基己基)酯(DEHP)不同污染程度的水稻土上盆栽不同品种通菜,应用GC／MS联机检测技术研究了通菜-土壤系统中DEHP的环境行为与归宿．结果表明,通菜中DEHP的含量为0．335～12．710mg·kg^-1,与土壤的污染程度呈正相关．不同品种通菜之间对DEHP的吸收累积效应存在显著差异,与其叶子大小存在一定的正相关关系．种植不同品种通菜后土壤中DEHP的残留量(1．424～19．834mg·kg^-1)存在显著差异．通菜对土壤中DEHP的BCFs都小于1．0,与土壤的污染程度呈负相关．不同通菜品种的BCFs之间存在显著差异,中等叶子通菜品种的BCFs相对较大。     

Integrated biosorption and photocatalytic oxidation treatment of di(2-ethylhexyl)phthalate

Di(2-ethylhexyl)phthalate (DEHP), a toxic phthalate ester, is a ubiquitous contaminant due to its extensive use and persistence. No available treatment method can cost-effectively remove it from industrial wastewater. In a previous study, DEHP was effectively removed from aqueous solution by adsorption onto the biomass of selected seaweed, i.e., beached seaweed and Sargassum siliquastrum. Since biosorption cannot detoxify DEHP, the degradation (and detoxification) of desorbed DEHP from seaweed biomass by photocatalytic oxidation (PCO) was attempted. The first part of this study was to optimize the conditions for the degradation of desorbed DEHP in aqueous solution by PCO. Under optimized conditions, a total degradation of 20 mg/L of DEHP was achieved within 45 min. The degradation intermediates/products such as phthalic anhydride and 2-ethyhexanol were identified by GC-MS analysis. Total organic carbon analysis was also used to ensure the complete mineralization of DEHP. The Microtox^® test was used to assess the toxicities of the parent and degraded compounds. In the second part of this study, DEHP was first removed and concentrated by adsorption onto seaweed biomass under the conditions optimized in the previous study. It was then desorbed from seaweed biomass and degraded by PCO. Results indicate that the treatment for DEHP by integrating biosorption and PCO is feasible.     

Isolation of Two Plasticizers,Bis(2-ethylhexyl) Terephthalate and Bis(2-ethylhexyl) Phthalate,from Capparis spinosa L. Leaves

Many plants have been known to be contaminated and accumulate plasticizers from the environment, including water sources, soil, and atmosphere. Plasticizers are used to confer elasticity and flexibility to various fiber and plastic products. Consumption of plasticizers can lead to many adverse effects on human health, including reproductive and developmental toxicity, endocrine disruption, and cancer. Herein, we report for the first time that two plasticizers, bis(2-ethylhexyl) terephthalate (DEHT) and bis(2-ethylhexyl) phthalate (DEHP), have been isolated from the leaves of Capparis spinosa L. (the caper bush), a plant that is widely used in food seasonings and traditional medicine. 297 mg/kg of DEHT and 48 mg/kg of DEHP were isolated from dried and grounded C. spinosa L. leaves using column chromatography and semi-preparative high-performance liquid chromatography. Our study adds to the increase in the detection of plasticizers in our food and medicinal plants and to the alarming concern about their potential adverse effects on human health.     

Effects of dietary di(2-ethylhexyl)phthalate on the structure and function of rat hepatocytes

The effect of the plasticizer di(2-ethylhexyl)phthalate on the intracellular membranes of hepatocytes was investigated. Supplementation of the diet with 2% plasticizer resulted in the appearance of a large number of peroxisomes, and the number of mitochondria was also greatly increased. No significant change in the amount or appearance of the endoplasmic reticulum was detected. The oxidation of palmitoyl-CoA in peroxisomes and the activities of carnitine-acyltransferases are increased to a great extent in both mitochondria and peroxisomes. Intact respiratory control and oxidative phosphorylation indicated that mitochondrial integrity was maintained during the induction. In microsomes, cytochrome P-450 and NADPH-cytochrome c reductase are elevated. The increased incorporation of glycerol into phospholipids indicated an increased rate of synthesis. The induction of peroxisomal and mitochondrial membranes and enzymes, but not of the membranes of the endoplasmic reticulum, by phthalate esters is an unusual and valuable induction pattern not seen with other inducers.