锯齿新米虾对Cu2+和毒死蜱毒性的生理响应

为研究水生生物对水体中重金属和有机磷农药毒性的生理响应,以锯齿新米虾为试验生物,采用半静止毒性试验方法,分别研究了Cu2+和毒死蜱5 d暴露时间对锯齿新米虾肌肉组织生理指标的影响.结果表明,锯齿新米虾暴露在不同浓度Cu2+和毒死蜱溶液下,随着暴露时间的延长,肌肉组织蛋白浓度均有不同程度的降低.Cu2+溶液较低浓度组(0.086 mg·L-1和0.172 mg·L-1)和毒死蜱较高浓度组(0.006 0μg·L-1和0.012 0μg·L-1)中,总SOD活性表现抑制效应;Cu2+溶液较高浓度组(0.344 mg·L-1和0.688 mg·L-1)表现为"抑制-诱导-抑制"的变化趋势,而毒死蜱较低浓度组(0.001 5μg·L-1和0.003 0μg·L-1)表现为"抑制-诱导"的变化趋势;暴露在Cu2+和毒死蜱溶液下,MDA含量变化相似,且在一定浓度范围内,随着Cu2+和毒死蜱浓度增加,MDA含量呈逐渐上升的趋势,说明Cu2+和毒死蜱加快了锯齿新米虾肌肉组织脂质过氧化作用;ACh E活性在Cu2+和毒死蜱溶液中表现为抑制效应,且在Cu2+和毒死蜱溶液的较高浓度组中,其活性随浓度的增加而呈逐渐降低的趋势,说明Cu2+和毒死蜱对锯齿新米虾肌肉组织的正常生理功能产生了影响,其浓度越高,产生的损伤效应越大.研究表明,鉴于不同浓度的Cu2+和毒死蜱溶液对锯齿新米虾肌肉组织的损伤程度不同,总SOD、MDA和ACh E可以作为评价重金属和有机磷农药对锯齿新米虾急性毒性效应的生理指标.     

磷酸三苯酯对斑马鱼早期生命阶段的神经毒性研究

磷酸三苯酯(triphenyl phosphate,TPP)作为多溴联苯醚类阻燃剂的替代产品,是一类生产和需求量均相当高的有机磷酸酯类阻燃剂,目前已在多种环境介质以及生物体内均有不同程度检出。由于结构和有机磷农药具有相似性,其对生物的神经毒性值得关注。本研究以斑马鱼为实验动物,研究了TPP(5~625%g·L-1)的胚胎发育毒性和行为毒性,并通过检测乙酰胆碱酯酶活性以及神经系统相关基因的转录水平,探讨其可能的毒性机制。研究发现,TPP可导致斑马鱼胚胎孵化时间延长,体长变短,心率变慢。同时,TPP暴露也可以影响斑马鱼幼鱼在持续光照和明暗周期刺激下的游泳行为,表现为低浓度增加而高浓度降低其游泳速度。而TPP暴露后幼鱼乙酰胆碱酯酶活性以及神经发育相关基因转录水平的变化可能是导致其行为毒性的原因。虽然实验中所设定的暴露浓度高于环境中一般浓度,但TPP在短期暴露中所表现出的胚胎和神经发育毒性表明TPP对于水生生物可能存在一定风险,需要进一步研究加以确认。     

In vitro effects of pesticides and metals on the activity of acetylcholinesterase (AChE) from different tissues of the blue mussel,Mytilus trossulus L.

The objective of this study was to conduct a comprehensive comparison of the effect on acetylcholinesterase (AChE) from various tissues of Mytilus trossulus caused by in vitro exposure to several pesticides and metals, because results available in the literature are inconsistent, difficult to compare, and sometimes contradict each other. For this purpose, fraction S10 extracted from gills, digestive gland, mantle and muscles, and the whole soft tissue of the mussel was exposed to several pesticides (dichlorvos, chlorpyrifos, fenitrothion, carbofuran and carbaryl) and metals (Cu, Zn, Cd, Hg, Pb) at a wide range of concentrations. AChE was inhibited in 50% or more in all the tissues exposed to dichlorvos, Cu, Hg and the mixture of Cu+Cd, and in some tissues exposed to carbaryl and carbofuran. The IC₅₀ was calculated where possible. No inhibition was found in the case of chlorpyrifos, Cd, Pb, and Zn.     

Deceased co-twin as a cause of false positive amniotic fluid AFP and AChE

A pregnancy was terminated because of persistently elevated amniotic fluid AFP (+10 S.D.) and an AChE band of low intensity on gel electrophoresis. No fetal anomalies were detected by ultrasonographic examination. Autopsy revealed an apparently normal fetus of about 20 weeks gestation. Attached to the placenta was a small sac containing a fetus papyraceus co-twin of about 8–9 weeks gestation. The small deceased co-twin and its gestational sac were not detected prenatally despite multiple ultrasonographic examinations. The difficulty in the interpretation of apparently conflicting results is emphasized.     

硫丹对草鱼乙酰胆碱酯酶及抗氧化酶活性的影响

研究了硫丹暴露对草鱼肝脏、肌肉抗氧化酶及脑乙酰胆碱酯酶活性的影响。结果表明,硫丹24 h暴露可诱导草鱼脑AChE活性,当暴露时间延长或质量浓度升高时,AChE活性表现为受抑制,120 h较高质量浓度组抑制率为41.8%和56.2%,抑制率与暴露时间呈良好的线性相关。硫丹暴露24 h后,草鱼肝脏及肌肉SOD、GSH-Px活性受到显著影响,表现出先诱导后缓慢降低的趋势,120 h后SOD活性显著低于对照组水平,GSH-Px活性与对照组无显著差异。在抗氧化酶受到影响的同时,鱼体脂质过氧化LPO程度不断上升,组织MDA含量逐渐增大,96 h达到最高值。硫丹可影响草鱼AChE及抗氧化酶活性,其变化可作为生物标志物,来评价化学污染物对水生动物的生物学效应。     

BDE-47和BDE-209对乙酰胆碱酯酶(AChE)活性的抑制特性及分子作用力研究

多溴联苯醚(PBDEs)是应用广泛的溴代阻燃剂,其中2,2',4,4'-四溴联苯醚(BDE-47)和2,2',3,3',4,4',5,5',6,6'-十溴联苯醚(BDE-209)广泛存在于环境中。PBDEs具有神经毒性,但其致毒机制尚不明确。本文通过研究BDE-47与BDE-209对乙酰胆碱酯酶(AChE)活性的影响及二者与AChE相互作用的光谱分析,揭示BDE-47与BDE-209导致神经毒性的致毒机制。BDE-47和BDE-209在一定浓度范围内均能够抑制AChE分解乙酰胆碱;随着浓度的增加,两者的抑制率均呈现出先增加后降低的规律。BDE-47浓度为400μmol·L-1时抑制率达到最大,为22.3%;BDE-209浓度为200μmol·L-1时抑制率达到最大,为11.2%。相同浓度下,BDE-47对AChE的抑制率始终大于BDE-209,表明AChE对BDE-47更加敏感。荧光光谱分析结果表明BDE-47和BDE-209与AChE之间的相互作用均主要为疏水作用,同时不存在范德华引力作用;BDE-47与AChE的结合常数大于BDE-209与AChE的结合常数,表明BDE-47更易与AChE相互作用。此外,温度的升高不利于BDE-47和BDE-209与AChE之间相互作用。BDE-47和BDE-209抑制AChE活性很可能是导致神经毒性通路之一。     

对有机磷农药敏感的海鱼脑AChE筛选研究

以10种海洋鱼类:鲅鱼、黄花鱼、鲈鱼、黄鱼、美国红鱼、真鲷、黑鲷、面包鱼、黑头鱼和鳗鲡为实验材料,采用半抑制浓度(IC₅₀)和双分子速率常数(K_i)为指标,通过体外抑制作用比较了这10种海鱼脑AChE对马拉硫磷和甲基对硫磷的敏感性,并分析了海鱼脑AChE作为监测海水中有机磷农药指示酶的可行性,由此筛选出适于制备海水有机磷农药酶传感器的指示酶.实验表明:10种海鱼脑AChE的基础活性在4.62～35.65μmol/(min·g)之间,面包鱼的酶活性最高,黄花鱼的酶活性最低.对马拉硫磷,由IC₅₀所得到的AChE敏感性为:鲅鱼、黄鱼、鲈鱼、真鲷>黑鲷>黑头鱼、鳗鲡>美国红鱼、黄花鱼>面包鱼;由Ki所得到的AChE敏感性为:鲅鱼>黄鱼、鲈鱼、真鲷>黑鲷、黑头鱼、鳗鲡>美国红鱼、黄花鱼>面包鱼.对甲基对硫磷,由IC50所得到的AChE敏感性为:鲅鱼>鲈鱼>黄鱼、黄花鱼、真鲷、黑鲷、黑头鱼、鳗鲡>美国红鱼>面包鱼;由Ki所得到的AChE敏感性为:鲅鱼>鲈鱼、黄鱼、黄花鱼、黑鲷、黑头鱼、鳗鲡>真鲷>美国红鱼>面包鱼.鲅鱼脑AChE对2种有机磷农药都较为敏感.影响鱼脑AChE对有机磷农药敏感性的主要因素为生物来源、农药种类和抑制方式,而与酶的基础活性无关.海鱼脑AChE活力抑制作用与水体中的有机磷农药具有良好的剂量-效应关系,在1～50mg/L浓度范围内,两者间呈现较好的线性相关性(r²≥0.93).根据AChE的敏感性及其对有机磷农药的线性响应程度,鲅鱼脑AChE适于作为监测马拉硫磷和甲基对硫磷酶传感器的指示酶.     

采用Caco-2和PC12细胞检测有机磷农药的细胞毒性

建立了采用哺乳动物细胞系Caco-2和PC12细胞检测有机磷农药细胞毒性的新方法.10μmol/L甲基对硫磷、敌敌畏、氧乐果、辛硫磷处理经过单层培养的Caco-2细胞,随着农药处理时间的延长,Caco-2细胞跨上皮细胞电阻抗值逐渐降低,处理60min时,辛硫磷和甲基对硫磷对Caco-2细胞具有明显的透过性,约是正常组的15倍,4种农药对Caco-2细胞透过性由大到小的顺序为:甲基对硫磷,辛硫磷,敌敌畏,氧乐果;001μmol/L的甲基对硫磷和辛硫磷使PC12细胞乙酰胆碱酯酶活性迅速降低,随着处理浓度的增加酶活性降低不明显;而小于10μmol/L的氧乐果和敌敌畏呈现渐进降低的趋势,当浓度大于10μmol/L时酶活性降低比较显著.      

Propetamphos enhances chlorpyrifos-induced organophosphate-induced delayed neurotoxicity (OPIDN) in Hens: a role for plasma butyrylcholinesterase

This study examined dose-response effects of chlorpyrifos (CPF) and propetamphos (PRO) on producing organophosphorus-induced delayed neurotoxicity (OPIDN) in hens. A single oral dose of 1, 3, 30, or 100 mg/kg of each compound was administered individually, or in combination to groups of 10 hens. Plasma butyrylcholinesterase (BChE) activity was inhibited by all doses of each individual compound. Chlorpyrifos alone or in combination with PRO resulted in decreased acetylcholinesterase (AChE) activity. No dose of CPF or PRO alone resulted in the inhibition of neuropathy target esterase (NTE) or produced OPIDN. In contrast, combined exposure to high doses of CPF and PRO produced diminished NTE activity and produced OPIDN. Brain NTE activity was decreased by 67% and 71% by combined treatment of 100 mg/kg CPF with 30 mg/kg and 100 mg/kg PRO, respectively, consistent with the development of ataxia and paralysis, characteristic of OPIDN. Only the highest dose of CPF 100 mg/kg interacted with the two highest doses of PRO to block NTE and induce OPIDN, whereas no marked interaction occurred at lower doses. Data indicate that PRO enhances the ability of high doses of CPF to produce OPIDN by binding to plasma BChE that appears to act as a buffering system to modulate the toxicity of CPF. In effect, the ability of BChE to bind to CPF is diminished by PRO, resulting in more CPF delivery to the brain, and subsequent development of OPIDN at a dose that does not initiate this effect by itself.     

蔬菜中有机磷农药残留的生物传感器法检测

将乙酰胆碱酯酶(AChE)固定在醋酸纤维膜上,将该膜组装在一支pH玻璃电极敏感膜表面,制成用于测定有机磷农药的电位型生物传感器,优化其制备方法。当溶液中存在有机磷农药,酶的活性受到抑制,酶活性的降低引起pH玻璃电极电位的改变,其改变程度与溶液中农药的浓度大小有关。用本文方法测定了蔬菜中的对硫磷、辛硫磷、氧乐果、敌敌畏,结果满意。