發(fā)布時(shí)間：2018-05-04 18:25

  本文選題：1-溴丙烷 + 神經(jīng)元特異性烯醇化酶��； 參考：《中國疾病預(yù)防控制中心》2017年碩士論文

【摘要】：目的1-溴丙烷(1-bromopropane,簡(jiǎn)稱1-BP)是一種無色至淡黃色有強(qiáng)烈特殊氣味的液體烷基溴化物,該物質(zhì)對(duì)大氣臭氧層無顯著破壞性,廣泛用于精密儀器清洗劑、噴霧黏合劑和脫脂劑等生產(chǎn)領(lǐng)域。我國現(xiàn)已成為全球1-BP的主要生產(chǎn)國之一,僅2008年全國產(chǎn)量就已達(dá)到20000噸,隨著其產(chǎn)量和使用量逐年遞增,接觸1-BP的從業(yè)人員也逐漸增多,1-BP的健康危害也越來越受到人們關(guān)注。職業(yè)暴露1-BP主要影響神經(jīng)、生殖以及肝臟等系統(tǒng),其中尤以神經(jīng)毒性最為明顯。以暴露、效應(yīng)和易感性標(biāo)志組成的生物標(biāo)志群不僅可以揭示1-BP健康損傷的過程和路徑,而且可以將預(yù)防和干預(yù)的節(jié)點(diǎn)得以提前。本研究從整體動(dòng)物實(shí)驗(yàn)和職業(yè)人群流行病學(xué)調(diào)查入手,通過對(duì)雄性Wistar大鼠染毒,觀察其中毒體征和神經(jīng)損傷特異性指標(biāo):神經(jīng)元特異性烯醇化酶(Neuron Specific Enolase,簡(jiǎn)稱NSE)、星形膠質(zhì)源性蛋白(S-100β protein,簡(jiǎn)稱S-100β)和環(huán)氧化酶-2(Cyclooxygenase-2,簡(jiǎn)稱COX-2)在大腦皮層和血清中的變化,以及尿中1-BP和代謝產(chǎn)物N-乙�；�-S-(正丙基)-L-半胱氨酸(N-Acetyl-S-(n-Propyl)-L-Cysteine,簡(jiǎn)稱AcPrCys)等暴露標(biāo)志的濃度監(jiān)測(cè),為探討毒效應(yīng)機(jī)制和進(jìn)一步的人群流行病學(xué)研究提供一定的理論支撐。通過對(duì)有代表性的1-BP生產(chǎn)和使用企業(yè)開展橫斷面流行病學(xué)調(diào)查,檢測(cè)作業(yè)場(chǎng)所中1-BP的暴露濃度,對(duì)暴露和對(duì)照人群開展問卷調(diào)查和職業(yè)健康監(jiān)護(hù),檢測(cè)其血清神經(jīng)損傷特異性指標(biāo)和尿中1-BP接觸標(biāo)志物的變化趨勢(shì),分析實(shí)驗(yàn)動(dòng)物和人群中相同指標(biāo)的關(guān)聯(lián)性,初步探索職業(yè)暴露人群可能的暴露和效應(yīng)標(biāo)志。方法1.動(dòng)物實(shí)驗(yàn)研究:選取健康雄性Wistar大鼠54只,隨機(jī)分為0(對(duì)照組)、500和1000ppml-BP暴露組,每組18只,采用口鼻吸入染毒方法,每天染毒6小時(shí)(8:00am-2:00pm),連續(xù)染毒21天,每天于三個(gè)時(shí)間點(diǎn)(10:00am、12:00am和2:00pm)取樣進(jìn)行濃度監(jiān)控。染毒期間隔天稱量并記錄動(dòng)物體重,于染毒-后第7、14和21天分批次各處死6只動(dòng)物,分離大腦皮層,采集血液并分離血清,每組兩只動(dòng)物分離大腦、垂體、脊髓、坐骨神經(jīng)和脛腓神經(jīng)用于病理檢測(cè),處死動(dòng)物前一天用代謝籠收集尿液。用ELISA試劑盒檢測(cè)大鼠大腦皮層和血清NSE、S-100β、COX-2等指標(biāo)的變化;GC-MS檢測(cè)尿中1-BP濃度;LC-MS/MS檢測(cè)尿中代謝產(chǎn)物AcPrCys濃度;2.現(xiàn)場(chǎng)流行病學(xué)調(diào)查:選取3家1-BP生產(chǎn)企業(yè)和3家1-BP使用企業(yè)的71名工人作為1-BP暴露組,選取從事餐飲、服裝等不接觸1-BP和相關(guān)物質(zhì)的71名工人為對(duì)照組,進(jìn)行暴露和對(duì)照組的問卷調(diào)查,調(diào)查人群的基本情況、職業(yè)史和健康情況等內(nèi)容,監(jiān)測(cè)現(xiàn)場(chǎng)1-BP的環(huán)境和個(gè)體暴露濃度,采集暴露和對(duì)照組的血并分離血清、尿液,并對(duì)暴露組進(jìn)行了神經(jīng)行為學(xué)測(cè)試和神經(jīng)科體檢。結(jié)果1.構(gòu)建1-BP對(duì)大鼠神經(jīng)的損傷模型:與對(duì)照組相比,500和1000ppm染毒21天的暴露組動(dòng)物體重增長(zhǎng)明顯減慢(P0.05),病理檢查可見小腦局部浦肯野細(xì)胞萎縮、腰髓灰質(zhì)空泡變性、脛腓神經(jīng)部分神經(jīng)纖維腫脹增粗;2.1-BP對(duì)大鼠神經(jīng)損傷特異性效應(yīng)標(biāo)志的影響:a)NSE:cNSE含量在染毒第1周至第3周,1000ppm組顯著高于對(duì)照組和500ppm組,sNSE含量在染毒第1周至第3周,500ppm組顯著高于對(duì)照組和 1000ppm 組(P0.05);b)S-100β:cS-100β含量在染毒第1周至第3周,1000ppm組顯著高于對(duì)照組和500ppm組,sS-100β含量在染毒第3周,1000ppm組顯著高于對(duì)照組和 500ppm 組(P0.05);c)COX-2:cCOX-2含量在染毒第1周至第3周,500和1000ppm組顯著高于對(duì)照組,且在染毒第2周和第3周,500ppm組顯著高于1000ppm組,sCOX-2含量在染毒第2周500ppm組顯著高于對(duì)照組,染毒第1周至第2周1000ppm組顯著高于對(duì)照組(P0.05);在大鼠大腦皮層和血清COX-2含量的變化具有相關(guān)性;3.1-BP暴露標(biāo)志的篩選:通過對(duì)尿中1-BP和代謝產(chǎn)物AcPrCys的檢測(cè),可見這兩種物質(zhì)均為1-BP特異的暴露標(biāo)志;500ppm大鼠血清NSE和COX-2含量變化與尿中AcPrCys濃度變化具有相關(guān)性;4.現(xiàn)場(chǎng)流行病學(xué)調(diào)查:暴露人群與對(duì)照人群相比,血清NSE、S-100β和COX-2的含量未見明顯差異;1-BP使用企業(yè)的暴露濃度普遍高于生產(chǎn)企業(yè),個(gè)別使用企業(yè)的暴露濃度超過國家標(biāo)準(zhǔn)(21mg/m3)。與較低暴露濃度的生產(chǎn)企業(yè)相比,較高暴露濃度的使用企業(yè)工人消極情緒得分和心境狀態(tài)總分顯著增高(P0.05)。暴露人群尿中1-BP可部分檢出,AcPrCys全部檢出,而在對(duì)照人群中上述兩種指標(biāo)則為陰性;AcPrCys與作業(yè)場(chǎng)所1-BP暴露濃度具有良好的相關(guān)性,可作為1-BP暴露標(biāo)志。結(jié)論1.綜合實(shí)驗(yàn)動(dòng)物大腦皮層和血清神經(jīng)特異性損傷指標(biāo)(NSE、S-100β和COX-2)的變化以及病理檢查結(jié)果,1-BP大鼠染毒模型可以在現(xiàn)有的實(shí)驗(yàn)條件和染毒條件下,引起實(shí)驗(yàn)動(dòng)物的亞急性神經(jīng)損傷;大腦皮層和血清COX-2存在關(guān)聯(lián)性;2.職業(yè)暴露人群中NSE、S-100β和COX-2變化不顯著,未發(fā)現(xiàn)不同暴露濃度人群在上述效應(yīng)標(biāo)志之間的差異性;3.在實(shí)驗(yàn)動(dòng)物和職業(yè)暴露人群的尿液中均可檢出1-BP和AcPrCys,且在實(shí)驗(yàn)動(dòng)物和人群中AcPrCys均與外暴露濃度具有良好的相關(guān)性,提示AcPrCys較1-BP更適合成為其暴露標(biāo)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.

【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R114

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李宏玲;劉浩中;宋向榮;陳曉燕;謝植偉;趙娜;蔡婷峰;梁e，

本文編號(hào)：1844101