毒死蜱低剂量重复染毒致学习记忆功能障碍及CREB信号途径的抑制(Repeated Exposures to Low-Level Chlorpyrifos Result in Learning Memory Impairment and Inhibition of the CREB Signal System)

毒死蜱(CPF)是我国推荐使用的低毒有机磷农药之一,目前在全世界广泛应用.选择不引起动物出现全身系统毒性的CPF剂量(1、5、10mg·kg-1)对大鼠连续染毒4周,评价CPF对动物的学习记忆功能和CREB信号通路的影响.结果显示:CPF连续染毒后动物出现记忆能力的损伤,随着染毒剂量的增加,海马、皮质和纹状体CREB及ERKI/Ⅱ磷酸化蛋白表达逐渐减少,这表明ERK/CREB信号通路可能是参与毒死蜱低剂量重复暴露致学习记忆功能改变的重要途径.     

克百威预处理对毒死蜱神经细胞毒性的影响

研究了氨基甲酸酯农药克百威预处理对有机磷农药毒死蜱神经细胞毒性的影响. 采用噻唑蓝(MTT)法测定克百威对原代培养皮层神经细胞存活率的影响,找出无细胞毒性浓度;采用无细胞毒性浓度的克百威对培养细胞进行不同时间的预处理后,加入毒死蜱(克百威仍存在)染毒48 h测定细胞毒性的变化;采用Western blot方法测定克百威不同时间预处理对毒死蜱激活ERK1/2作用的影响. 结果表明:10 μmol/L的克百威处理72 h显示无细胞毒性,10 μmol/L的克百威预处理(2～24 h)可减轻毒死蜱的毒性,其中预处理8 h时作用最强;克百威预处理降低了毒死蜱对ERK1/2的激活作用,抑制ERK1/2激活作用与细胞毒性减轻相关. 克百威预处理时间过短(0～2 h),对毒死蜱的细胞毒性及ERK1/2激活影响很小. 克百威预处理一段时间后可拮抗毒死蜱的细胞毒性,该作用机制与抑制ERK1/2激活有关;但克百威预处理时间过短,则不产生拮抗作用. 提示氨基甲酸酯农药与有机磷农药之间的联合作用模式受二者染毒时间间隔的影响.      

邻苯二甲酸二乙基己酯(DEHP)对THP-1细胞IL-1β和MMP-8表达及ROS的影响

为探讨邻苯二甲酸二乙基己酯((DEHP)的细胞免疫毒性作用与机制,采用RT-PCR和ELISA方法,考察了0.05～1μmol·L-1浓度范围内的DEHP对THP-1细胞白细胞介素-1β(IL-1β)及基质金属蛋白酶-8(MMP-8)基因和蛋白表达的影响;采用免疫印迹WesternBlot方法检测DE-HP对ERK1/2磷酸化水平的影响;以2,'7-'二氯荧光素二乙酸酯(DCFH-DA)为荧光探针检测1～50μmol·L-1DEHP对细胞内活性氧(ROS)产生的影响。结果显示,0.05和0.2μmol·L-1DEHP在6h内显著诱导IL-1β和MMP-8基因表达(P<0.05或0.01);0.05～1μmol·L-1DEHP刺激细胞48h,可诱导IL-1β蛋白表达,并表现出明显的剂量-效应关系,线性拟合的确定系数为0.937;0.05μmol·L-1DEHP刺激细胞6或12h,显著诱导MMP-8蛋白表达(P<0.05);0.2μmol·L-1DEHP在15～30min内快速诱导ERK1/2磷酸化;1～50μmol·L-1DEHP浓度依赖性刺激细胞中ROS的产生;ERK/MAPK抑制剂PD98059显著抑制DEHP诱导的MMP-8分泌,但对IL-1β分泌未表现出抑制作用。研究表明,DEHP可能是经ERK/MAPK信号路径诱导MMP-8基因和蛋白的表达,经其他路径诱导IL-1β基因和蛋白的表达,诱导细胞内ROS的产生,从而激发炎症反应,进而损害免疫系统功能引发哮喘等炎症性疾病。此研究结果可为DEHP的暴露风险评估提供参考。     

Tumor promoting effects of cyanobacterial extracts are potentiated by anthropogenic contaminants--evidence from in vitro study

Inhibition of gap junctional intercellular communication (GJIC) is affiliated with tumor promotion process and it has been employed as an in vitro biomarker for evaluation of tumor promoting effects of chemicals. In the present study we investigated combined effects of anthropogenic environmental contaminants 2,2′,4,4′,5,5′-hexachlorobiphenyl (PCB 153) and fluoranthene, cyanotoxins microcystin-LR and cylindrospermopsin, and extracts of laboratory cultures of cyanobacteria Aphanizomenon gracile and Cylindrospermopsis raciborskii, on GJIC in the rat liver epithelial cell line WB-F344. Binary mixtures of PCB 153 with fluoranthene and the mixtures of the two cyanobacterial strains elicited simple additive effects on GJIC after 30 min exposure, whereas microcystin-LR and cylindrospermopsin neither inhibited GJIC nor altered effects of PCB 153 or fluoranthene. However, synergistic effects were observed in the cells exposed to binary mixtures of anthropogenic contaminants (PCB 153 or fluoranthene) and cyanobacterial extracts. The synergistic effects were especially pronounced after prolonged (6-24 h) co-exposure to fluoranthene and A. gracile extract, when mixture caused nearly complete GJIC inhibition, while none of the individual components caused any downregulation of GJIC at the same concentration and exposure time. The effects of cyanobacterial extracts were independent of microcystin-LR or cylindrospermopsin, which were not detected in cyanobacterial biomass. It provides further evidence on the presence of unknown tumor promoting metabolites in cyanobacteria. Clear potentiation of the GJIC inhibition observed in the mixtures of two anthropogenic contaminants and cyanobacteria highlight the importance of combined toxic effects of chemicals in complex environmental mixtures.