焦炉作业工人职业暴露估测及其端粒损伤生物标探索

发布时间：2018-05-21 12:06

  本文选题：焦炉逸散物 + 基因多态　； 参考：《郑州大学》2016年博士论文

【摘要】：焦炉逸散物(coke oven emissions,COEs)可引起职业性肺癌,我国尚未制定其职业接触生物限值。近年来端粒DNA损伤在职业卫生领域研究中发展迅速,有必要探讨其作为COEs效应标志的价值,以及端粒损伤过程中相关基因的作用。目的1.探讨焦炉工人环境暴露剂量和尿代谢物含量以及端粒损伤等生物标志价值,通过剂量-反应关系与基准剂量研究为职业卫生标准的修订提供依据。2.结合职业暴露人群端粒通路基因、细胞周期调控基因和代谢酶基因多态性及基因表达情况,探讨相关易感性标志及相关因素对端粒损伤的影响。方法1研究对象COEs暴露组由工龄在1年以上的544名焦化厂作业人员组成,对照组由238名非职业性有毒有害物质暴露的健康人群组成。问卷调查研究对象的姓名、性别、出生日期、吸烟、饮酒和职业史等基本情况。2研究方法2.1暴露检测:对车间空气中COEs浓度进行检测并估算其累积暴露剂量。采用HPLC法进行班末尿中1-羟基芘、3-羟基菲和2-羟基萘浓度测定。2.2效应标志检测:采集研究对象外周血,应用q PCR的技术测定外周血白细胞DNA端粒长度,试剂盒测定血清丙二醛、过氧化氢、过氧化氢酶和总抗氧化能力。2.3基因检测:采用PCR与限制性片段长度多态性方法进行端粒通路基因、细胞周期调控基因和代谢酶基因多态性检测。采用q PCR的方法检测相关基因m RNA表达。3统计学分析采用SPSS21.0统计软件进行数据分析,非正态分布数据用M(P25,P75)进行统计描述,组间比较用秩和检验,分类变量组间比较采用?2检验。采用stata13.0统计软件对端粒长度的影响因素进行多因素分位数回归分析。检验水准α=0.05。基准剂量分析采用美国环境保护局发布的BMDS 2.6.0.1软件。结果1.焦炉工职业暴露调查暴露COEs时间加权平均浓度(Time weighted average concentration,TWA)超标的工种主要有装煤车司机、拦焦车司机、焦侧炉门工、上升管工和热修工。COEs暴露组累积暴露剂量为1.12(0.34,2.14)mg/m3·年,尿1-羟基芘和3-羟基菲浓度分别为83.80(40.29,163.55)pg/μg肌酐和18.20(9.99,35.63)pg/μg肌酐,影响因素为性别、年龄、吸烟、饮酒和工龄,2-羟基萘浓度为84.97(46.49,174.45)pg/μg肌酐,主要影响因素为性别、年龄和吸烟。2.焦炉工外周血DNA端粒长度和氧化损伤效应探讨暴露组和对照组端粒长度分别为0.75(0.51,1.08)和1.05(0.76,1.44),两组差异有统计学意义(Z=7.692,P0.001);暴露组与对照组的总抗氧化能力为分别为11.96(8.88,14.68)和14.55(10.51,19.24),差异有统计学意义(Z=6.614,P0.001)。氧化损伤各指标与端粒长度无明显相关性。基准剂量(Benchmark dose,BMD)研究显示:外暴露COEs浓度的BMDL值为1.839 mg/m3·年,尿代谢物1-羟基芘BMDL值为43.254(pg/μg肌酐),3-羟基菲BMDL值为11.369(pg/μg肌酐)。3.焦炉工端粒长度与基因多态性和相关m RNA表达的关系暴露组CYP2E1基因rs3813867位点CC基因型个体端粒长度为0.97(0.70,1.20),长于GG型个体的端粒长度0.73(0.50,1.09),差异有统计学意义(Z=2.536,P=0.011)。暴露组P21、TPP1、TRF1和TRF2基因m RNA表达低于对照组(P0.05)。暴露组P53和POT1的m RNA表达高于对照组(P0.05)。P53、POT1、TIN2和TRF2基因m RNA表达与端粒长度均呈负相关(P0.05),TRF1基因m RNA表达与端粒长度呈正相关(P=0.015)。COEs暴露组POT1基因rs10250202位点AA基因型个体的POT1基因m RNA表达低于对照组(P=0.041);COEs暴露组P21基因rs1801270和rs1059234位点的各基因型个体的P21基因m RNA表达均低于对照组(P0.05)。COEs暴露组P53基因rs1042522、rs1625895位点中的各基因型个体和rs17878362位点SS基因型个体的P53基因m RNA表达均高于对照组(P0.05);COEs暴露组TRF1基因rs3863242位点中的各基因型个体的TRF1基因m RNA表达均低于对照组(P0.001)。4.焦炉工端粒长度影响因素的多因素分析在第15th~85th分位数下暴露分组因素与端粒长度呈负相关。性别因素与第75th分位数下的端粒长度呈正相关。年龄与第50th分位数下的端粒长度呈负相关。暴露浓度与第5th~25th分位数下的端粒长度呈负相关。TERT基因rs2736098位点多态性与第75th分位数下的端粒长度呈正相关。P53基因rs17878362位点多态性与第75th~95th分位数下的端粒长度呈正相关。CYP2E1 rs3813867位点多态性与第25th分位数下的端粒长度呈正相关。TPP1基因m RNA表达量与第75th和85th分位数下的端粒长度呈正相关。TRF1基因m RNA表达量与第75th分位数下的端粒长度呈负相关。P53基因m RNA表达量与第75th分位数下的端粒长度呈负相关。RAP基因m RNA表达量与第15th分位数下的端粒长度呈负相关。TIN2基因m RNA表达量与第85th分位数下的端粒长度呈负相关。结论1.外周血白细胞DNA端粒长度和尿代谢物中1-羟基芘可能作为焦炉工人效应生物标志和暴露生物标志,采用BMD方法获得的COEs和1-羟基芘的BMDL可为COEs车间空气浓度限值和职业接触生物限值的制定提供依据。2.单因素分析发现,CYP2E1 rs3813867位点多态对外周血白细胞端粒DNA长度有影响,可能是端粒长度的易感性生物标志。端粒长度的缩短可能与P53、POT1、TIN2和TRF2 m RNA表达量增高及TRF 1 m RNA表达降低有关。3.分位数回归方法探讨端粒长度影响因素:暴露浓度,P53、RAP、TIN2和TRF1等基因m RNA的表达升高可能为端粒长度的危险因素。TPP1基因m RNA表达升高,TERT基因rs2736098位点AA基因型、P53基因rs17878362位点的SL基因型、CYP2E1基因rs3813867位点CC基因型可能是端粒长度的保护因素。
[Abstract]:Coke oven emissions (COEs) can cause occupational lung cancer. In recent years, our country has not formulated its occupational exposure biological limit. In recent years, telomere DNA damage has developed rapidly in the field of occupational health research. It is necessary to explore its value as a marker of COEs effect and the role of related genes in the process of telomere injury. 1. objective to discuss the coke oven The value of environmental exposure dose, urinary metabolite content and telomere damage, and to provide a basis for the revision of the occupational health standards through the dose response relationship and the baseline dose study. The.2. combined with the telomere pathway gene of the occupational exposure population, the cell cycle regulation gene and the metabolite gene polymorphism and gene expression, The effect of related susceptibility markers and related factors on telomere injury. Method 1 COEs exposure group was composed of 544 workers working in 544 coking plants for more than 1 years. The control group was composed of 238 healthy people exposed to non occupational toxic and harmful substances. The questionnaire survey was conducted on the names, sex, date of birth, smoking, drinking and the study. Occupational history and other basic cases.2 research method 2.1 exposure testing: the concentration of COEs in the air of the workshop was detected and the cumulative exposure dose was estimated. The HPLC method was used to measure the 1- hydroxy pyrene in the end of the class urine, the concentration of 3- hydroxy phenanthrene and 2- hydroxy naphthalene was detected for the detection of the.2.2 effect, and the peripheral blood of the study subjects was collected, and the peripheral blood was measured by the technique of Q PCR Cell DNA telomere length, detection of serum malondialdehyde, hydrogen peroxide, catalase and total antioxidant capacity.2.3 gene detection: using PCR and restriction fragment length polymorphism method to carry out telomere pathway gene, cell cycle regulation gene and metabolic enzyme gene polymorphism detection. Q PCR method was used to detect the related gene m RNA expression.3 Statistical analysis uses SPSS21.0 statistical software for data analysis, non normal distribution data is statistically described with M (P25, P75), groups are compared with rank sum test, and the classification variables are compared to 2 test. Multifactor quantile regression analysis is used to analyze the influence factors of telomere length by stata13.0 statistical software. Test level alpha =0.05. basis. The BMDS 2.6.0.1 software issued by the American environmental protection agency was used for the quasi dose analysis. Results 1. coke oven workers exposed the COEs time weighted mean concentration (Time weighted average concentration, TWA) over the standard of the coal truck driver, the chauffeur driver, the coke oven door worker, the rising pipe worker and the hot repair worker,.COEs exposure group. The exposure dose of 1.12 (0.34,2.14) mg/m3 year, urine 1- hydroxy pyrene and 3- hydroxy phenanthrene concentration were 83.80 (40.29163.55) pg/ Mu creatinine and 18.20 (9.99,35.63) pg/ micron g creatinine respectively. The factors were sex, age, smoking, drinking and working age, 2- hydroxy naphthalene concentration was 84.97 (46.49174.45), the main factors were sex, age and smoking coke. The telomere length and oxidative damage effect of DNA in the peripheral blood were 0.75 (0.51,1.08) and 1.05 (0.76,1.44) in the exposure group and the control group, respectively. The two groups were statistically significant (Z=7.692, P0.001), and the total antioxidant capacity of the exposure group and the control group was 11.96 (8.88,14.68) and 14.55 (10.51,19.24), respectively (Z). =6.614, P0.001). There was no significant correlation between the indexes of oxidative damage and the length of telomere. The study of Benchmark dose (BMD) showed that the BMDL value of COEs concentration in external exposure was 1.839 mg/m3. The 1- hydroxyl pyrene BMDL value of urine metabolite was 43.254 (pg/ micronic creatinine), and the hydroxyl phenanthrene value of 11.369 (creatinine) coke oven telomere length and gene polymorphism The relationship between sex and related M RNA expression in the CYP2E1 gene rs3813867 locus of the exposed group was 0.97 (0.70,1.20), which was longer than the telomere length 0.73 (0.50,1.09) of the GG type, and the difference was statistically significant (Z=2.536, P=0.011). The expression of RNA was higher than that of the control group (P0.05).P53, and the expression of M RNA in the POT1, TIN2 and TRF2 genes was negatively correlated with the telomere length (P0.05). The TRF1 gene m RNA expression was positively correlated with the telomere length. The expression of P21 gene m RNA in each genotype of 70 and rs1059234 loci was lower than that of the control group (P0.05).COEs exposure group P53 gene rs1042522, and the P53 genes of each genotype and SS genotype at the rs1625895 locus were higher than those of the control group. The expression of M RNA of TRF1 gene in type individuals was lower than that of the control group (P0.001).4. coke oven telomere length influencing factor analysis. The exposure group factor was negatively correlated with the telomere length under the 15th~85th quantile. The gender factor was positively correlated with the telomere length under the 75th quantile. The telomere length under the year of the year and the 50th quantile was negative. Correlation. The exposure concentration and the telomere length under the 5th~25th quantile were negatively correlated with the.TERT gene rs2736098 polymorphism and the telomere length under the 75th quantile, the.P53 gene rs17878362 locus polymorphism was positively correlated with the telomere length under the 75th~95th quantile and the.CYP2E1 rs3813867 locus polymorphism and the 25th quantile The telomere length is positively related to the m RNA expression of the.TPP1 gene and the telomere length under the 75th and 85th quantiles. The expression of the.TRF1 gene m RNA is negatively correlated with the telomere length under the 75th quantile, and the m RNA expression is negatively correlated with the telomere length under the digits. The negative correlation.TIN2 gene m RNA expression was negatively correlated with the telomere length under the 85th digits. Conclusion the DNA telomere length of the peripheral blood leukocyte and the 1- hydroxy pyrene in the urinary metabolite may be the biomarker and the exposure biomarker of the coke oven workers. The BMDL of COEs and 1- hydroxy pyrene obtained by BMD formula can be COEs. .2. single factor analysis showed that CYP2E1 rs3813867 locus polymorphisms had an influence on the telomere length of peripheral blood leukocytes and may be an susceptibility biomarker of telomere length. The shortening of telomere length may be associated with the increased expression of P53, POT1, TIN2 and TRF2 m RNA and TRF 1 m. RNA expression reduced.3. quantile regression to explore the influence factors of telomere length: exposure concentration, P53, RAP, TIN2 and TRF1, the increase in the expression of M RNA may be a risk factor for telomere length:.TPP1 gene m RNA expression. The 7 locus CC genotype may be a protective factor for telomere length.
【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R135
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本文编号：1919118