Primary neuronal-astrocytic co-culture platform for neurotoxicity assessment of di-(2-ethylhexyl) phthalate

Plastics such as polyvinyl chlorides （PVC） are widely used in many indoor constructed environments; however, their unbound chemicals, such as di-（2-ethylhexyl） phthalates （DEHP）, can leach into the surrounding environment. This study focused on DEHP＇s effect on the central nervous system by determining the precise DEHP content in mice brain tissue after exposure to the chemical, to evaluate the specific exposure range. Primary neuronal-astrocyte co-culture systems were used as in vitro models for chemical hazard identification of DEHP. Oxidative stress was hypothesized as a probable mechanism involved, and therefore the total reactive oxygen species （ROS） concentration was determined as a biomarker of oxidative stress. In addition, NeuriteTracer, a neurite tracing plugin with ImageJ, was used to develop an assay for neurotoxicity to provide quantitative measurements of neurological parameters, such as neuronal number, neuron count and neurite length, all of which could indicate neurotoxic effects. The results showed that with 1 nmol/L DEHP exposure, there was a significant increase in ROS concentrations, indicating that the neuronal-astrocyte cultures were injured due to exposure to DEHP. In response, astrocyte proliferation （gliosis） was initiated, serving as a mechanism to maintain a homeostatic environment for neurons and protect neurons from toxic chemicals. There is a need to assess the cumulative effects of DEHP in animals to evaluate the possible uotake and effects on the human neuronal system from exoosure to DEHP in the indoor environment.     

α7nAChR在氯化甲基汞神经毒性中的作用

为探讨环境染污染物氯化甲基汞(methylmercury chloride,Me Hg Cl)的神经毒性作用机制,采用MTT、免疫细胞荧光、dot Blot、qRT-PCR等实验技术检测Me Hg Cl对PC12细胞活性及其α7亚型烟碱型胆碱能受体(α7 nicotinic acetylcholine receptor,α7n AChR)蛋白和mRNA表达水平的影响。结果显示Me Hg Cl抑制PC12细胞活性,显著降低α7n AChR的蛋白和mRNA的表达水平,呈浓度依赖性。上述结果表明,Me Hg Cl对神经细胞毒性作用可能与抑制α7n AChR表达有关。     

DEHP暴露对鲤鱼肾脑生物标志物的影响

为探讨邻苯二甲酸二乙基己酯(DEHP)对鱼类肾、脑组织生物标志物的影响,将鲤鱼分别在浓度为5、20、80、160 mg·L~(-1)的DEHP水体,暴露20 d,以水和吐温-80为对照,测定肾脑组织中多种酶活性和丙二醛(MDA)含量的变化。结果表明:与水对照组相比,吐温-80组所测各项指标,差异均不显著。与水和吐温-80对照组相比,各染毒组,肾脏中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和抗羟自由基、抗超氧阴离子的活力均显著降低(P0.05或P0.01)。谷胱甘肽过氧化物酶(GSH-Px)酶活性呈现先升高后降低,在5、20、80 mg·L~(-1)浓度组显著升高(P0.05)。丙二醛(MDA)含量在80、160 mg·L~(-1)浓度下,显著升高(P0.05),但脑组织中变化不显著。脑中一氧化氮合酶(NOS)在各浓度组显著升高(P0.05或P0.01);乙酰胆碱酯酶(ACh E)在80、160 mg·L~(-1)时降低显著(P0.05);钙调神经磷酸酶(CaN)呈先升高后降低的趋势,在20 mg·L~(-1)下,升高显著(P0.01),在80、160 mg·L~(-1)浓度组,降低极显著(P0.01);抗羟自由基、抗超氧阴离子活性均显著下降(P0.05或P0.01)。可见,在实验浓度范围内,DEHP对鲤鱼具有一定免疫毒性和神经毒性,进一步的机理有待研究。     

苯并(a)芘神经毒性研究进展

苯并(a)芘(BaP)是一种广泛存在于环境中的多环芳烃,具有致癌、致畸、致突变性。目前BaP的神经毒性研究零散而不深入,本文主要总结分析了BaP的神经毒性表现,简要介绍了BaP诱发神经毒性呈现的剂量-效应关系,进一步系统阐述了目前研究中发现的可能分子机制,包括BaP诱导神经递质及其代谢物含量的改变,相关DNA及蛋白质的损伤,抗氧化系统及线粒体的改变等,以期为进一步研究提供参考。     

甲状腺激素脱碘酶在孕哺期十溴联苯醚(BDE 209)暴露致子代小鼠神经发育毒性中的潜在作用

研究孕哺期BDE209暴露对母鼠胎盘和子代脑组织甲状腺激素脱碘酶(deiodinase,DI)基因表达的影响,及其在子鼠神经发育毒性效应中的作用。将75只雌性昆明小鼠随机分为对照组、低剂量组和高剂量组,暴露BDE20910 d后,与雄鼠合笼,每组选取怀孕时间相近(相差不过2 d)的8只母鼠孕期持续染毒至子鼠断乳。采用实时荧光定量PCR检测孕17~18 d胎盘、出生后60 d子鼠脑组织3种类型脱碘酶基因相对表达;利用Morris水迷宫评价出生后60 d子鼠学习记忆能力;测量第2、16、30和60天子鼠体重,观察孕哺期BDE 209暴露对子代生长发育的作用。结果显示,BDE 209对子鼠出生时体重未见明显影响;出生后30 d,高剂量BDE 209暴露组雌性子鼠体重显著低于对照组子鼠体重(p0.05),而低、高剂量暴露组雄性子鼠体重均显著低于对照组子鼠体重(p0.05,p0.01);出生后60 d,BDE 209对子鼠体重影响不明显。BDE 209暴露能够显著延长出生后60 d雌、雄性子鼠逃避潜伏期(p0.05或p0.01)。BDE209暴露显著降低母鼠胎盘中3种类型脱碘酶(主要是DI-3)基因表达(p0.05或p0.01);同时,BDE 209暴露可诱导出生60 d后雄性子鼠脑组织中DI-1基因表达(p0.05或p0.01),抑制雄性子鼠脑组织中DI-3基因表达(p0.05);BDE 209暴露对出生后60 d雌鼠脑组织脱碘酶未见明显影响(p0.05)。研究结果表明,孕哺期BDE209暴露可能通过影响母鼠胎盘组织脱碘酶(特别是DI-3)基因表达,导致子鼠神经发育毒性效应-学习记忆能力障碍。     

铅和得克隆联合暴露对斑马鱼胚胎的神经毒性作用

以斑马鱼(Danio rerio)为研究对象,探讨铅(Pb)、得克隆(DP)及二者联合急性暴露对斑马鱼胚胎的神经毒性作用。结果表明,Pb(5、20μg·L~(-1))和DP(15、60μg·L~(-1))单独暴露均会引起斑马鱼自主运动频率增加,触摸反应能力和自由游泳活力下降,并且抑制初级运动神经元的生长,加剧尾部细胞凋亡。但与20μg·L~(-1)Pb单独暴露相比,高剂量联合暴露(20μg·L~(-1)Pb+60μg·L~(-1)DP)使斑马鱼的自主运动频率显著降低(P0.05),触摸反应能力和自由游泳活力显著增强(P0.05),初级运动神经元轴突长度显著增加(P0.05),尾部细胞凋亡减少。与5μg·L~(-1)Pb单独暴露相比,低剂量联合暴露(5μg·L~(-1)Pb+15μg·L~(-1)DP)也显著减少斑马鱼尾部的细胞凋亡(P0.05)。上述结果表明,Pb或DP单独暴露对斑马鱼均可引起神经毒性作用;但二者联合暴露对斑马鱼自主运动、触摸反应以及自由游泳活力的影响则表现为拮抗作用。     

内质网应激介导的凋亡在PBDE-47致大鼠海马损伤中的作用

为了明确内质网应激(endoplasmic reticulum stress,ERS)介导的凋亡在2,2’,4,4’-四溴联苯醚(2,2’,4,4’-tetrabromodiphenylether,PBDE-47)致大鼠神经毒性中的作用,在脑发育的突增期(出生第10天),分别用0 mg·kg~(-1)、1 mg·kg~(-1)、5 mg·kg~(-1)和10mg·kg~(-1)PBDE-47进行单次灌胃染毒,并从出生第8天开始每天给予150 mg·kg~(-1)ERS抑制剂4-苯基丁酸(phenylbutyric acid,PBA),持续3周。仔鼠出生第8周末,从对照组、10 mg·kg~(-1)PBDE-47处理组、150 mg·kg~(-1)PBA处理组和150 mg·kg~(-1)PBA+10mg·kg~(-1)PBDE处理组中随机选取8只大鼠进行水迷宫实验。然后将所有动物断头处死,分离大鼠海马组织,观察其海马组织形态学改变、测定ERS标志分子(GRP78、IRE1和CHOP)和凋亡相关蛋白Cyt c的表达水平。结果显示,10 mg·kg~(-1)PBDE-47处理可导致雌性大鼠海马CA4区细胞排列紊乱、锥形神经细胞数量减少甚至消失,尼氏小体数量减少,大鼠逃避潜伏期延长(P0.05)。5和10 mg·kg~(-1)PBDE-47处理可显著上调ERS相关蛋白IRE1和CHOP的表达水平(P0.05),并明显增强海马组织凋亡相关蛋白Cyt c的表达。PBA干预可明显降低PBDE-47诱导的IRE1和CHOP以及Cyt c蛋白的表达水平,提示PBDE-47可通过ERS介导凋亡,导致大鼠海马组织损伤,从而影响大鼠学习记忆功能。     

Introduction

Abstract

Eight pairs of O‐methyl and O‐ethyl O‐(substituted‐phenyl) phenylphosphonothionates were evaluated with respect to their delayed neurotoxic activity in hens. O‐methyl compounds were in all cases more active than their O‐ethyl analogs. The neurotoxic potential of the O‐methyl phenylphosphonothionates was 2,5‐diCl >4‐NO₂ >2,4,5‐triCl and 2,4,6‐triCl >2,4‐diCl >2,5‐diCl‐4‐Br >4‐CN, when single oral doses were given. Both EPN‐ethyl and leptophos‐raethyl were more neurotoxic in multiple dermal than multiple oral dosing regimens. LD₅₀s for mice and flies were established.     

An autism cohort study of cobalt levels following vitamin B12 injections

The effects of methylcobalamin (vitamin B12) injection (75 µg kg^?1) on cobalt levels in autism spectrum disorders (ASDs) and potential toxic effects of cobalt on human neurons were evaluated. A cohort of ASDs (n = 72) presenting to the Genetic Centers of America were evaluated for the frequency of methylcobalamin injections and cobalt levels using Laboratory Corporation of America (LabCorp) testing. Potential toxic effects of cobalt (cobalt(II) nitrate hexahydrate) to human neurons grown in vitro were evaluated. Methylcobalamin injections significantly increased the mean levels of plasma cobalt (6.83-fold) and urinary cobalt (51-fold) in comparison to unexposed subjects, and significant positive correlations were found between the frequency of methylcobalamin injections and the levels of plasma and urinary cobalt (injections every second day induced cobalt levels in excess of the LabCorp cobalt occupational maximum exposure limits). The LC₅₀ in human neurons following 24 h incubation with cobalt is 559 µM. Research must be conducted to determine optimal therapeutic methylcobalamin doses.     

Effects of repeated pesticide exposure on the peripheral and central nervous systems

This study was designed to compare organic farmers (comparison group) and pesticide-using farmers (pesticide group) in terms of neurobehavioral performance and sensorimotor function. The subjects were recruited in October 2004 from Chungnam Province and the neighboring Jeolla Province in western South Korea. Psychological assessment was carried out using the Wechsler Adult Intelligence Scale, the Mini-Mental State Examination, and the Bender Gestalt test. Neuroselective sensory nerve conduction threshold was measured at both left and right middle fingers by using an electrodiagnostic device to determine current perception thresholds (CPTs). Three independent CPT measurements were obtained from each test site by using sinusoidal stimuli at three frequencies. CPT testing showed no difference between the two groups. No significant differences were found in the large myelinated (2000?Hz stimulation; cutaneous touch, pressure), small myelinated (250?Hz stimulation; mechanoreceptive, pressure, temperature, fast pain) and small unmyelinated (5?Hz stimulation; polymodal nociceptive, temperature, slow pain, postganglionic sympathetic) C-fibers. Furthermore, there was no difference found between the pesticide and comparison groups in psychological variables, including intelligence quotient, orientation, memory, calculation, similarities, visual sensitivity, and psychomotor speed. In conclusion, chronic pesticide exposure has no apparent effect on either the peripheral or central nervous system in Korean farmers.