氧化型染发剂对沙蚕的毒性效应及对部分酶活性的影响

在实验室模拟条件下,研究了氧化型染发剂对双齿围沙蚕(Perinereis aibuhiteusis Grube)的急性毒性效应及不同暴露浓度(0、40、80、160、320mg·L-1)和暴露时间(0、4、6、8、10d)下对其体内乙酰胆碱酯酶(AChE)和超氧化物歧化酶(SOD)活性的影响.结果表明,1)暴露3d后氧化型染发剂对沙蚕的半数致死浓度(LC50)为1849.6mg·L-1;2)沙蚕体内AChE和SOD活性受到氧化型染发剂暴露浓度的显著影响:暴露3d后,随暴露浓度的升高(0～320mg·L-1),AChE和SOD活性均呈先被诱导后逐渐下降的变化趋势;3)沙蚕体内AChE和SOD活性也受到暴露时间的显著影响:40mg·L-1浓度下,随暴露时间的延长(0～10d),SOD活性呈先下降又缓慢上升的趋势,而AChE活性变化没有显著规律;4)通过比较发现,相对于AChE,SOD的活性变化更能反映氧化型染发剂对沙蚕的毒性作用.     

Acetylcholinesterase activity is affected by stress conditions in<Emphasis Type="Italic"> Paracentrotus lividus</Emphasis> coelomocytes

The presence and function of acetylcholinesterase (AChE) in blood cells and plasma is well known. The enzyme activity is associated to red blood cell membranes, as well as to the perinuclear region of leucocytes, suggesting a role in the function of the latter. Recently coelomocytes, acknowledged progenitors of immune cells in vertebrate systems, have been shown to respond to stress conditions, both in laboratory experiments and field studies. Bearing in mind the similarity between coelomocytes and vertebrate blood cells, we investigated the presence of AChE activity in coelomocytes of the sea urchin Paracentrotus lividus characterized by specific inhibitors, in relation to cold stress induced in the laboratory. Exposure to stress was followed by electrophoretic analysis of AChE activity and Western blot analysis of AChE-like immunoreactivity. Moreover, quantitative analysis of the different molecular forms of AChE was carried out with the aid of a spectrophotometric method. The effects of stress on AChE activity were time dependent. In fact, the enzyme activity was detected by the catalytic activity in cleaving the substrate acetyl thiocholine iodide, which appears to be formed by different isoforms of the enzyme, compared to that of Electrophorus, which was used as a control. Western blot analyses showed the presence of different immunoreactive molecules, with molecular mass ranging from 265 kDa (corresponding to Electrophorus molecules) to about 180 kDa. According to quantitative analysis, the isoforms that were differentiated corresponded to an AChE with the capacity to cleave acetyl--methyl thiocholine iodide (the so-called true AChE, E.C. 3.1.1.7, corresponding to the 265 kDa band) and to a ChE with the capacity to cleave propionyl thiocholine iodide as well. The latter is an ancestral form of AChE, mainly present in invertebrates. Each of these forms was affected differently by varying time exposures.Communicated by R. Cattaneo-Vietti, Genova     

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。     

Carbofuran-induced effects on acetylcholinesterase (AChE) in erythrocytes and liver of Cyprinus carpio

A study was conducted to determine the effects of long-term exposure to carbofuran on erythrocyte and liver acetylcholinesterase (AChE) activity in Cyprinus carpio, and assess which tissue was more sensitive to pesticide exposure examining this enzyme. Fish were allowed to accilimatize in toxicant-free water for 24 days after 60-day exposure. AChE activity was determined with a spectrophotometer using acetylthiocholine iodide as a substrate in erythrocytes and liver. Enzymic activity of erythrocytes and liver decreased over time. A higher degree of enzyme inhibition was observed in erythrocyte compared to liver. The degree of enzyme inhibition was positively correlated with exposure time. When exposed fish were transferred to clean water, recovery was greater in erythrocytes than liver. Erythrocyte AChE activity recovered after 18 days, while it required 21 days in liver. The findings of this study indicate that erythrocyte AChE is more sensitive to carbofuran than liver. The greater sensitivity of erythrocyte AChE suggests that it may be more useful as a biomarker for monitoring status of pollution in freshwater bodies.     

太阳鱼脑乙酰胆碱酯酶的分离纯化

采用硫酸铵沉淀–普鲁卡因胺亲和柱层析方法,对太阳鱼(Lepomis gibbosus)脑部乙酰胆碱酯酶分离纯化,纯化倍数为234倍,回收率为25.67%,经SDS-PAGE电泳检测,呈现单一的条带,相对分子质量约为68 000,表明其纯度达到电泳纯.     

Inhibitory effect of arachidonic acid on venom acetylcholinesterase

The venom of krait (Bungarus sindanus), an Elapidae snake, contains high level of acetylcholinesterase (AChE) activity. The effect of arachidonic acid on krait (Bungarus sindanus) venom AChE (acetylcholine hydrolase, EC 3.1.1.7) was studied using kinetic methods. This study evaluated for the first time whether arachidonic acid acts as potential AChE inhibitor of krait venom. Kinetic studies, using Line-Weaver Burk and Dixon plots, indicated that the inhibition produced by arachidonic acid was non-competitive, i.e., both k_m and V _max fell with increase in the concentration of arachidonic acid. The concentration of arachidonic acid required to inhibit 50% of enzymatic activity was found to be 1.9?µM while the inhibitory constant (K_i ) was 1.4?µM. The present results suggest that essential fatty acid (arachidonic acid) may also be used as an inhibitor of snake venom AChE.     

三丁基氯化锡对红鳍笛鲷的急性毒性及不同组织生化指标的影响

为研究有机锡化合物对海洋生物抗氧化防御系统和神经系统的影响,以我国东南沿海常见的中下层增养殖鱼类红鳍笛鲷(Lutjanus erythopterus)幼鱼为实验生物,采用半静态毒性实验方法,分别研究了三丁基氯化锡(tributyltin chloride,TBTCl)的急性毒性和96 h暴露时间内对红鳍笛鲷幼鱼鳃、脑和...     

三苯基锡和五氯酚胁迫斑马鱼生理生化的影响

以斑马鱼(Brachy danio rerio)作为受试生物,分析不同暴露浓度、不同暴露时间三苯基锡(TPT)、五氯酚(PCP)对斑马鱼生理生化指标的影响,在0,1.05,2.09,4.18,6.27,8.36μg·L-1的TPT和0,5.01,10.02,20.04,40.08,80.16μg·L-1的PCP暴露下,...     

六种农药对乙酰胆碱酯酶活性的体外毒性效应

通过体外毒性试验,研究乙酰甲胺磷、巴丹、高效氯氰菊酯、甲基硫菌灵、异菌脲和哒螨灵6种农药对乙酰胆碱酯酶活性的毒性效应。结果表明,在体外反应体系中, 6种农药均对乙酰胆碱酯酶活性有明显的抑制作用,其中乙酰甲胺磷对乙酰胆碱酯酶活性抑制率随着浓度增高先下降后上升, 1mg·L-1时抑制率最低,为10. 07%;甲基硫菌灵对乙酰胆碱酯酶活性抑制率随着浓度增高先上升后下降, 0. 125mg·L-1时抑制率最高,达60. 32%;巴丹、高效氯氰菊酯、异菌脲、哒螨灵对乙酰胆碱酯酶活性抑制率随浓度增高而上升。     

Toxicological attributes of plant chemicals and their biochemical impacts on cholinesterase and protein levels in relation with conventional insecticides against mosquito larvae of Karachi city

In order to control mosquito, different chemicals were applied against the larvae of Culex fatigans (Diptera Culicidae). The tested chemicals were Lambda cyhalothrin, chlorpyrifos, Azadirachta indica (Spindales meliaceae) neem extract, and Sueda monoica extract. All chemicals were found to have significant efficacy against Culex fatigans mosquito larvae. In terms of efficacy, the biochemical effects after treatment with such chemicals were found to vary. Total protein levels and acetylcholinesterase (AChE) activity were the parameters examined. Chlorpyrifos, Lambda cyhalothrin, and neem elevated AChE activity levels whereas the plant extract S. monoica decreased enzymic activity. Total protein levels were decreased by Lambda cyhalothrin and chlorpyrifos whereas protein was increased by neem extract and S. monoica.     

Use of Mytilus galloprovincialis and Tapes philippinarum as sentinel organisms for the development of a biosurveillance program in the Pialassa Baiona coastal lagoon (Ravenna, Italy)

The Pialassa Baiona is an intertidal lagoon located north from the town of Ravenna (Italy). It shows a certain degree of contamination caused by past uncontrolled discharges, and we aim at developing a bioassessment program to be used for future monitoring and conservation plans. A battery of six biomarkers (lysosome membrane stability, lipofuscin and neutral lipid accumulation, hsp70 expression, metallothionein levels and acetylcholinesterase (AChE) activity) was investigated in Mytilus galloprovincialis transplanted for 30 days in different sites. Tapes philippinarum are widely distributed within the lagoon, and we explored the possibility to use resident clams for future biosurveillance programs. Adenylyl cyclase (ACase) activity was also assessed in tissues of both bivalves as a future possible biomarker. Taken together, our results indicate that mussels are suitable sentinel organisms also within such a peculiar environment; alteration of four biomarkers was observed in tissues of mussels transplanted in two sites out of the five analyzed, indicating their low water quality. T. philippinarum showed a barely detectable AChE activity and absence of neutral lipids in digestive glands, therefore appeared unsuitable for our purpose, at least using the selected battery of biomarkers. ACase activity was not different among animals collected from the different sites.     

Ameliorated effects of garlic (Allium sativum) on biomarkers of subchronic acrylamide hepatotoxicity and brain toxicity in rats

Acrylamide (ACR) exerts its toxicity through stimulation of the oxidative stress; yet, its effect on neurotransmitter catabolic enzymes has not been elucidated. We investigated the effects of ACR exposure on brain and hepatic tissues antioxidant enzymes activities and different markers such as, acetylcholinesterase (AChE), nitric oxide (NO), monoamine oxidase (MAO), and lipid profile, and to evaluate the protective effects of garlic against ACR toxicity. Male Sprague-Dawley rats were exposed to ACR (1 mg kg^?1 body weight) with or without diet containing 1.5% of garlic powder for 40 days. ACR administration showed a decrease in AChE activity associated with an increase in MAO activity in both brain and hepatic tissues. In addition, ACR administration increased the lipid peroxidation and NO levels of both tissues while decreased the activities of glutathione (GSH), superoxide dismutase, and glutathione-S-transferase (GST). On the other hand, the activities of glutathione peroxidase (GPx) and catalase activities increased as a consequence of GSH depletion after ACR exposure. Finally, ACR exposure increased the brain and liver lipid profile of cholesterol, triglycerides and total lipid, while phospholipids level was decreased. Coadministration of garlic powder with ACR significantly attenuated oxidative stress, MAO activity, and inflammation in brain and hepatic tissues but did not ameliorate AChE activity. In conclusion, our results emphasized the role of garlic as a potential adjuvant therapy to prevent ACR neurotoxicity and hepatotoxicity.     

Acute toxicity evaluation of triazophos,deltamethrin and their combination on earthworm,Eudrilus eugeniae and its impact on AChE activity

The acute toxicity of three formula grade pesticides namely, triazophos (an organophosphate, OP), deltamethrin (a pyrethroid) and combined pesticide (triazophos?+?deltamethrin) was determined in earthworm, Eudrilus eugeniae. They were exposed to different concentrations of these pesticides for 48 h by paper contact toxicity method. The LC₅₀ values for triazophos, deltamethrin and of combination were determined as 0.076, 0.031 and 0.065?μg/cm², respectively. To study the sublethal effect of these pesticides, E. eugeniae were exposed to 5% and 10% of LC₅₀ of pesticides for 48 h. Morphological alterations such as coiling, clitellar swelling, mucus release and bleeding followed by body segmentation were observed in exposed earthworm. Acetylcholinesterase (AChE) activity assayed in different regions of body segment exhibited a significant (p?<?0.05) decrease in its activity particularly in the pre-clitellar region as compared to other regions. The altered behavioural responses in pesticides exposed earthworms would have been due to decline in AChE activity of the nervous system.     

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。     

三氯杀螨醇对泥鳅血清GST和AChE活力的影响

生化指标能更灵敏地反映环境污染的状况和危害程度,日益成为环境领域研究的热点.为研究三氯杀螨醇对泥鳅生化指标的影响,将泥鳅暴露于含不同剂量三氯杀螨醇的人工沉积物中,在暴露24、48和96 h后采集泥鳅血液样本,测定血清中谷胱甘肽硫转移酶(GST)和乙酰胆碱酯酶(AChE)的活力.结果表明:泥鳅在人工沉积物中暴露24和96...     

3种有机磷农药对水生生物的乙酰胆碱酯酶抑制效应的物种敏感度分析初探

通过筛选敌敌畏、马拉硫磷和对硫磷3种有机磷农药对水生生物的急性毒性数据和乙酰胆碱酯酶抑制效应数据,构建物种敏感度分布曲线进行了比较分析。结果表明,敌敌畏对水生生物的急性毒性和乙酰胆碱酯酶抑制效应的大小顺序为:酶体内抑制效应酶体外抑制效应急性毒性;马拉硫磷和对硫磷的乙酰胆碱酯酶抑制效应数据不足但趋势相似,顺序为:酶体内抑制效应急性毒性酶体外抑制效应。敌敌畏的急性毒性和酶体外抑制效应的5%危害浓度(HC5)分别为2.07μg·L~(-1)和1.53μg·L~(-1),两者相差1.4倍。在水质基准推导中,乙酰胆碱酯酶抑制效应数据对有机磷农药的水生生物基准具有重要的参考价值。     

Effect of repeated oral administration of bifenthrin antioxidant status and acetylcholinesterase activity in brain of rats

The oxidative-stress-inducing potential of the pyrethroid, bifenthrin, in the brain of rats was evaluated following daily oral administration of 5.8 mg/kg body weight. Lipid peroxidation in the brain was increased, while the activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase decreased. The activity of acetylcholinesterase (AChE) in erythrocytes and brain also decreased. Bifenthrin induces severe oxidative stress in brain, and inhibits brain and erythrocyte AChE.     

Neurotoxicity and oxidative stress induced by permethrin in gills of the freshwater mussel Unio ravoisieri

Pyrethroids are contaminants found in the aquatic environment, and their toxicological effects on aquatic organisms have received extensive attention. However, the impact on freshwater bivalve of exposure to these chemicals is still largely unknown. Freshwater mussels Unio ravoisieri were exposed to two nominal permethrin (PM) concentrations C1?=?50?µg/L and C2?=?100?µg/L during 7 days. The measured concentrations of PM using gas chromatography (GC/ECD) in the treated aquariums were, respectively, 28.7–62.3?µg/L. Catalase (CAT), Glutathione-S-transferase (GST), and Acetylcholinesterase (AChE) activities, Glutatione (GSH) and Malondialdehyde (MDA) levels were determined in gills of U. ravoisieri. Significant increase in CAT activity by the lowest concentration and decrease by highest concentration were observed. Additionally, GST activity was increased in a concentration-dependent manner. However, statistically significant decrease in GSH levels (about 39%) was observed only at high concentration of this compound (100?µg/L). PM generated an increase in MDA levels reaching the highest value at the high concentration. AChE activity of mussel ranging from 51% inhibition at lowest concentration 50?µg/L to 89% inhibition at highest concentration 100?µg/L. The results indicated that oxidative stress and cell damage might be one of the main mechanisms of PM toxicity to freshwater mussels.     

Effect of fenvalerate technical grade on acetyl cholinesterase activity in Indian bullfrog Haplobatrachus tigerinus (Daudin)

Indian bullfrog Haplobatrachus tigerinus (Daudin) was exposed to sublethal dose (1/3 of LC50 I.E. 1.166 mg/kg) of fenvalerate technical grade and the effect was studied on the specific activity of acetyl cholinesterase in the different tissues of frog viz., brain, muscle, liver, kidney and testis at different time periods viz., 3,6, 12, 24, 48 and 72 hours. The inhibition of specific activity of acetyl cholinesterase was in the order of kidney > brain > muscle > liver > testis. A significant inhibition was noticed in kidney at 12 hours (-64.33%) and no effect was noticed at 3 hours in testis (+0.67%). The AChE activity was inhibited in first three hours of administration of fenvalerate in all the tissue tested. The inhibition continued upto 6 hours or 2 hours in different tissue but the recovery was started by 24 hours and almost completed by 72 hours.     

Nerve conduction studies in sprayers occupationally exposed to mixture of pesticides in a mango plantation at Lucknow,North India

In this study, nerve conduction, organochlorine (OC) pesticide residues in blood and cholinesterase activity levels of pesticide sprayers employed in mango plantations at Lucknow, North India, were determined. Fifty-two sprayers from mango plantations who regularly spray mixture of pesticides like organophosphates (OP), OCs, and carbamates were considered as exposed group. Eighteen subjects with similar socio-economic status of exposed group, who do not handle pesticides, were selected as controls. Questionnaire-based interviews related to personal and occupational histories of the study subjects were carried out. Sprayers did not use any personal protective equipment during pesticide handling. The blood-pesticide analyses of sprayers show higher mean values of hexachlorohexane (HCH), γ-HCH, δ-HCH, total HCH, op-DDT compared with the controls. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activites were significantly reduced among sprayers. Risk of motor nerve conduction deficits was observed in sprayers with low AChE activity. Negative correlation of motor and sensory nerve conduction deficits was observed with the duration of exposure and age among sprayers. The study demonstrated that the prolonged exposure to mixture of pesticides, ergonomic factors, decline in cholinesterase activity may lead to nerve conduction dysfunction. The findings suggest the need for controlled use of pesticides in the plantation and indicated the need for training and implementation of hygiene practices like proper usage of personal protective equipments.