CYP2E1及CYP2B6基因多态性对苯乙烯接触工人尿中代谢产物及DNA损伤影响的研究

发布时间：2018-05-15 01:25

本文选题：苯乙烯 + CYP2E1　；参考：《沈阳医学院》2017年硕士论文

【摘要】：目的本研究通过对苯乙烯作业工人基因多态性、淋巴细胞彗星实验、尿中8-羟基脱氧鸟苷(8-hydroxy-2'-deoxyguanosine,8-OHdG)含量及尿中代谢产物苯乙醇酸(mandelic acid,MA)和苯乙醛酸(phenylglyoxylic acid,PGA)的检测,探讨低浓度苯乙烯对作业工人尿中代谢产物和DNA损伤的影响,以及CYP2E1和CYP2B6基因多态性与尿中代谢产物及DNA损伤的相关性,确定苯乙烯敏感遗传损伤指标及易感性标志物。为保护工人健康及对苯乙烯接触易感人群筛选提供参考依据。方法选择有苯乙烯接触工龄在1年以上的工人50名为接触组,同一地区不接触苯乙烯及其它已知遗传毒物的工人48名为对照组。苯乙烯接触组与对照组相比在性别、年龄、工龄及吸烟比例上具有均衡可比性。通过填写调查问卷了解苯乙烯作业工人一般情况;采用高效液相色谱法(High performance liquid chromatography,HPLC)检测尿中代谢产物MA和PGA含量;采用酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测尿中8-OHdG含量,单细胞凝胶电泳实验(single cell gel eletrophoresis,SCGE)检测DNA链断裂情况;采用聚合酶链反应—限制性片段长度多态性(Polymerase Chain Reaction-Restriction Fragment Length Polymorphism,PCR-RFLP)技术检测 CYP2E1 及 CYP2B6 基因型的频率分布情况。结果苯乙烯作业工人尿中8-OHdG含量及彗星尾长均高于对照组,差异有统计学意义;携带CYP2E1野生纯合型(C1C1)个体尿中代谢产物MA+PGA含量及尿中8-OHdG含量均高于携带突变型(C1C2+C2C2)的个体,差异有统计学意义;携带CYP2E1不同基因型个体之间的彗星尾长差异无统计学意义;携带CYP2B6基因516GT位点和718AG位点的野生型与突变型个体之间的尿中代谢产物、8-OHdG含量及彗星尾长差异均无统计学意义。结论1.低浓度苯乙烯可导致作业工人DNA损伤。2.尿中8-OHdG可作为低浓度苯乙烯作业工人DNA氧化损伤的敏感标志物。3.在低浓度苯乙烯环境中,CYP2E1突变型基因降低了苯乙烯代谢的能力,携带CYP2E1野生型(C1C1)个体为易感性人群。4.CYP2B6两个突变位点的不同基因型在低浓度苯乙烯环境中均对苯乙烯代谢无明显影响。
[Abstract]:Objective to detect the gene polymorphisms of workers exposed to styrene, comet assay of lymphocytes, the content of 8-hydroxy-2guanosine (8-OHdG) in urine and the metabolites of phenylglycolate mandelic acid (MAA) and phenylglyoxylic acid (PGA) in urine. To investigate the effects of low concentration styrene on urinary metabolites and DNA damage in workers, and the relationship between CYP2E1 and CYP2B6 gene polymorphisms, urinary metabolites and DNA damage, and to determine the sensitive genetic damage index and susceptible markers of styrene. To provide a reference for the protection of workers' health and the screening of susceptible population to styrene exposure. Methods 50 workers with exposure to styrene and 48 workers without exposure to styrene and other known genetic poisons in the same area were selected as exposure group. The sex, age, length of service and smoking ratio in the styrene exposure group were balanced and comparable with those in the control group. To find out the general situation of styrene workers by filling out questionnaires; to detect the contents of MA and PGA in urine by high performance liquid chromatographyHPLC, and to detect 8-OHdG in urine by enzyme-linked immunosorbent assay-esa (Elisa), and to measure the content of MA and PGA in urine by high performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assayya Elisa (Elisa). Single cell gel eletrophoresistics (SCGE) was used to detect DNA strand breaks, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the frequency distribution of CYP2E1 and CYP2B6 genotypes. Results the contents of 8-OHdG and comet tail length in urine of workers exposed to styrene were significantly higher than those of the control group, and the contents of MA PGA and 8-OHdG in urine of individuals carrying CYP2E1 wild homozygous type C1C1) were higher than those of individuals carrying mutant C1C2C2C2). There was no significant difference in comet tail length among individuals with different genotypes of CYP2E1. There was no significant difference in urinary metabolite 8-OHdG content and comet tail length between wild-type and mutant individuals carrying 516GT and 718AG loci of CYP2B6 gene. Conclusion 1. Low concentration of styrene could cause DNA damage. 2. 8-OHdG in urine can be used as a sensitive marker of DNA oxidative damage in workers exposed to low concentration styrene. In low concentration styrene environment, CYP2E1 mutant gene reduced the ability of styrene metabolism. Individuals carrying CYP2E1 wild type C _ 1C _ 1) were susceptible population. 4. Different genotypes of CYP2B6 had no significant effect on styrene metabolism in low concentration styrene environment.
【学位授予单位】：沈阳医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R135

【参考文献】