1-溴丙烷典型生物标志研究

发布时间：2018-05-04 18:25

本文选题：1-溴丙烷 + 神经元特异性烯醇化酶　；参考：《中国疾病预防控制中心》2017年硕士论文

【摘要】：目的1-溴丙烷(1-bromopropane,简称1-BP)是一种无色至淡黄色有强烈特殊气味的液体烷基溴化物,该物质对大气臭氧层无显著破坏性,广泛用于精密仪器清洗剂、喷雾黏合剂和脱脂剂等生产领域。我国现已成为全球1-BP的主要生产国之一,仅2008年全国产量就已达到20000吨,随着其产量和使用量逐年递增,接触1-BP的从业人员也逐渐增多,1-BP的健康危害也越来越受到人们关注。职业暴露1-BP主要影响神经、生殖以及肝脏等系统,其中尤以神经毒性最为明显。以暴露、效应和易感性标志组成的生物标志群不仅可以揭示1-BP健康损伤的过程和路径,而且可以将预防和干预的节点得以提前。本研究从整体动物实验和职业人群流行病学调查入手,通过对雄性Wistar大鼠染毒,观察其中毒体征和神经损伤特异性指标:神经元特异性烯醇化酶(Neuron Specific Enolase,简称NSE)、星形胶质源性蛋白(S-100β protein,简称S-100β)和环氧化酶-2(Cyclooxygenase-2,简称COX-2)在大脑皮层和血清中的变化,以及尿中1-BP和代谢产物N-乙酰基-S-(正丙基)-L-半胱氨酸(N-Acetyl-S-(n-Propyl)-L-Cysteine,简称AcPrCys)等暴露标志的浓度监测,为探讨毒效应机制和进一步的人群流行病学研究提供一定的理论支撑。通过对有代表性的1-BP生产和使用企业开展横断面流行病学调查,检测作业场所中1-BP的暴露浓度,对暴露和对照人群开展问卷调查和职业健康监护,检测其血清神经损伤特异性指标和尿中1-BP接触标志物的变化趋势,分析实验动物和人群中相同指标的关联性,初步探索职业暴露人群可能的暴露和效应标志。方法1.动物实验研究:选取健康雄性Wistar大鼠54只,随机分为0(对照组)、500和1000ppml-BP暴露组,每组18只,采用口鼻吸入染毒方法,每天染毒6小时(8:00am-2:00pm),连续染毒21天,每天于三个时间点(10:00am、12:00am和2:00pm)取样进行浓度监控。染毒期间隔天称量并记录动物体重,于染毒-后第7、14和21天分批次各处死6只动物,分离大脑皮层,采集血液并分离血清,每组两只动物分离大脑、垂体、脊髓、坐骨神经和胫腓神经用于病理检测,处死动物前一天用代谢笼收集尿液。用ELISA试剂盒检测大鼠大脑皮层和血清NSE、S-100β、COX-2等指标的变化;GC-MS检测尿中1-BP浓度;LC-MS/MS检测尿中代谢产物AcPrCys浓度;2.现场流行病学调查:选取3家1-BP生产企业和3家1-BP使用企业的71名工人作为1-BP暴露组,选取从事餐饮、服装等不接触1-BP和相关物质的71名工人为对照组,进行暴露和对照组的问卷调查,调查人群的基本情况、职业史和健康情况等内容,监测现场1-BP的环境和个体暴露浓度,采集暴露和对照组的血并分离血清、尿液,并对暴露组进行了神经行为学测试和神经科体检。结果1.构建1-BP对大鼠神经的损伤模型:与对照组相比,500和1000ppm染毒21天的暴露组动物体重增长明显减慢(P0.05),病理检查可见小脑局部浦肯野细胞萎缩、腰髓灰质空泡变性、胫腓神经部分神经纤维肿胀增粗;2.1-BP对大鼠神经损伤特异性效应标志的影响:a)NSE:cNSE含量在染毒第1周至第3周,1000ppm组显著高于对照组和500ppm组,sNSE含量在染毒第1周至第3周,500ppm组显著高于对照组和 1000ppm 组(P0.05);b)S-100β:cS-100β含量在染毒第1周至第3周,1000ppm组显著高于对照组和500ppm组,sS-100β含量在染毒第3周,1000ppm组显著高于对照组和 500ppm 组(P0.05);c)COX-2:cCOX-2含量在染毒第1周至第3周,500和1000ppm组显著高于对照组,且在染毒第2周和第3周,500ppm组显著高于1000ppm组,sCOX-2含量在染毒第2周500ppm组显著高于对照组,染毒第1周至第2周1000ppm组显著高于对照组(P0.05);在大鼠大脑皮层和血清COX-2含量的变化具有相关性;3.1-BP暴露标志的筛选:通过对尿中1-BP和代谢产物AcPrCys的检测,可见这两种物质均为1-BP特异的暴露标志;500ppm大鼠血清NSE和COX-2含量变化与尿中AcPrCys浓度变化具有相关性;4.现场流行病学调查:暴露人群与对照人群相比,血清NSE、S-100β和COX-2的含量未见明显差异;1-BP使用企业的暴露浓度普遍高于生产企业,个别使用企业的暴露浓度超过国家标准(21mg/m3)。与较低暴露浓度的生产企业相比,较高暴露浓度的使用企业工人消极情绪得分和心境状态总分显著增高(P0.05)。暴露人群尿中1-BP可部分检出,AcPrCys全部检出,而在对照人群中上述两种指标则为阴性;AcPrCys与作业场所1-BP暴露浓度具有良好的相关性,可作为1-BP暴露标志。结论1.综合实验动物大脑皮层和血清神经特异性损伤指标(NSE、S-100β和COX-2)的变化以及病理检查结果,1-BP大鼠染毒模型可以在现有的实验条件和染毒条件下,引起实验动物的亚急性神经损伤;大脑皮层和血清COX-2存在关联性;2.职业暴露人群中NSE、S-100β和COX-2变化不显著,未发现不同暴露浓度人群在上述效应标志之间的差异性;3.在实验动物和职业暴露人群的尿液中均可检出1-BP和AcPrCys,且在实验动物和人群中AcPrCys均与外暴露浓度具有良好的相关性,提示AcPrCys较1-BP更适合成为其暴露标志。
[Abstract]:Objective 1- 1-bromopropane (1-BP) is a liquid alkyl bromide with a strong special odor of colorless to light yellow. This substance has no significant damage to the atmospheric ozone layer. It is widely used in precision instrument cleaning agents, spray adhesives and degreasing agents. China has now become one of the major producers of 1-BP in the world, only 200 The national output has reached 20000 tons in the past 8 years. As its output and use increase year by year, the workers exposed to 1-BP are increasing, and the health hazards of 1-BP are getting more and more attention. The occupational exposure 1-BP mainly affects nervous, reproductive and liver systems, especially the neurotoxicity is most obvious. The biomarker group composed of chronicles can not only reveal the process and path of 1-BP health damage, but also advance the prevention and intervention nodes. This study starts with the whole animal experiment and the epidemiological survey of the occupational population, and through the exposure to the male Wistar rats, the specific indexes of the toxic signs and nerve damage are observed. Specific enolase (Neuron Specific Enolase, NSE), astroglial protein (S-100 beta protein, S-100 beta) and cyclooxygenase -2 (Cyclooxygenase-2, COX-2) in the cerebral cortex and serum, as well as the 1-BP and metabolites of the metabolites of the metabolite (n-propyl) cysteine The concentration monitoring of exposure markers, such as e, AcPrCys, provides a theoretical support for exploring the toxic effect mechanism and further population epidemiological studies. Through a cross-sectional epidemiological survey of the representative 1-BP production and use enterprises, the exposure concentration of 1-BP in the workplace is detected, and a questionnaire is conducted for the exposed and controlled population. Survey and occupational health monitoring, detection of the specific index of nerve injury in serum and the change trend of 1-BP contact markers in urine, analysis the association of the same indexes in the experimental animals and the population, and initially explore the possible exposure and effect markers of the occupational exposure population. Method 1. animal test study: 54 healthy male rats were selected, and the random samples were selected randomly. The group was divided into 0 (control group), 500 and 1000ppml-BP exposure groups, 18 in each group, using oral and nasal inhalation, 6 hours (8:00am-2:00pm) every day for 21 days, at three time points (10:00am, 12:00am and 2:00pm) sampling for concentration monitoring. During the period of exposure, the weight of animals was recorded and recorded on 7,14 and 21 days after exposure. 6 animals were sacrificed each time to separate the cerebral cortex, collect blood and separate the serum. Two animals in each group separated brain, pituitary, spinal cord, sciatic nerve and tibiofibular nerve for pathological examination. The urine was collected by metabolic cage on the day before the animals were killed. The changes of NSE, S-100 beta, COX-2, and other indexes in the rat cerebral cortex and serum were detected by ELISA kit; GC-MS The concentration of 1-BP in urine was detected and the concentration of AcPrCys in urine was detected by LC-MS/MS; 2. field epidemiological survey: 71 workers of 1-BP production enterprises and 3 1-BP use enterprises were selected as 1-BP exposure group, and 71 workers engaged in food and beverage, clothing and other non 1-BP and related substances were selected as the control group, and the questionnaire of exposure and control group was carried out. Investigate the basic situation, occupational history and health status of the population, monitor the environment of 1-BP and the concentration of individual exposure, collect the blood of the exposed and control groups and separate the serum and urine, and carry out the neurobehavioral test and the neurology examination of the exposed group. Results the damage model of the 1. constructs 1-BP to the rat's nerve: compared with the control group, 5 The body weight increased significantly (P0.05) in the exposed group of 00 and 1000ppm for 21 days. Pathological examination showed the atrophy of the local Purkinje cells in the cerebellum, the degenerative of the gray matter in the lumbar pulp, the swelling of the partial nerve fibers of the tibia and fibula, and the effect of 2.1-BP on the specific effect of the nerve injury in rats: a) the content of NSE:cNSE was first weeks to third weeks, 1000p PM group was significantly higher than control group and 500ppm group, sNSE content was first weeks to third weeks, 500ppm group was significantly higher than control group and 1000ppm group (P0.05); b) S-100 beta: cS-100 beta content in first weeks to third weeks, 1000ppm group was significantly higher than the control group and 500ppm group, sS-100 beta content was third weeks, the group was significantly higher than the control group and the control group. Group (P0.05), c) COX-2:cCOX-2 content in first weeks to third weeks, 500 and 1000ppm groups were significantly higher than the control group, and in the second and third weeks, 500ppm group was significantly higher than the 1000ppm group, sCOX-2 content in the second week 500ppm group was significantly higher than the control group, first weeks to second weeks 1000ppm group significantly higher than the control group (P0.05); in the rat brain skin. There was a correlation between the changes of COX-2 content in the layer and serum; screening of 3.1-BP exposure markers: through the detection of 1-BP and metabolite AcPrCys in urine, these two substances were all 1-BP specific exposure markers; the changes of NSE and COX-2 content in serum of 500ppm rats were related to the variation of AcPrCys concentration in urine; 4. field epidemiological survey: exposure Compared with the control population, there was no significant difference in the content of serum NSE, S-100 beta and COX-2; the exposure concentration of 1-BP used enterprises was generally higher than that of the production enterprises, and the exposure concentration of individual enterprises exceeded the national standard (21mg/m3). Compared with the low exposed production enterprises, the higher exposure concentration used enterprise workers' negative emotional scores. The total score of the state of mental state increased significantly (P0.05). 1-BP could be detected partly in the urine of the exposed population, and all of the AcPrCys were detected, but the above two indexes were negative in the control population; AcPrCys had a good correlation with the 1-BP exposure concentration in the workplace, which could be used as a sign of 1-BP exposure. The changes in sexual injury index (NSE, S-100 beta and COX-2) and the results of pathological examination showed that the 1-BP rat model could cause subacute nerve injury in experimental animals in the existing experimental conditions and exposure conditions; there was a correlation between the cerebral cortex and the serum COX-2; the changes of NSE, S-100 beta and COX-2 in the 2. occupational exposure population were not significant, and no difference was found. 1-BP and AcPrCys were detected in 3. of the urine of experimental and occupational exposed populations, and AcPrCys in both experimental animals and population had a good correlation with external exposure, suggesting that AcPrCys was more suitable to be a sign of exposure than 1-BP.

【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R114

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