本文選題：焦?fàn)t逸散物 + 基因多態(tài)�。� 參考：《鄭州大學(xué)》2016年博士論文

【摘要】：焦?fàn)t逸散物(coke oven emissions,COEs)可引起職業(yè)性肺癌,我國尚未制定其職業(yè)接觸生物限值。近年來端粒DNA損傷在職業(yè)衛(wèi)生領(lǐng)域研究中發(fā)展迅速,有必要探討其作為COEs效應(yīng)標(biāo)志的價(jià)值,以及端粒損傷過程中相關(guān)基因的作用。目的1.探討焦?fàn)t工人環(huán)境暴露劑量和尿代謝物含量以及端粒損傷等生物標(biāo)志價(jià)值,通過劑量-反應(yīng)關(guān)系與基準(zhǔn)劑量研究為職業(yè)衛(wèi)生標(biāo)準(zhǔn)的修訂提供依據(jù)。2.結(jié)合職業(yè)暴露人群端粒通路基因、細(xì)胞周期調(diào)控基因和代謝酶基因多態(tài)性及基因表達(dá)情況,探討相關(guān)易感性標(biāo)志及相關(guān)因素對端粒損傷的影響。方法1研究對象COEs暴露組由工齡在1年以上的544名焦化廠作業(yè)人員組成,對照組由238名非職業(yè)性有毒有害物質(zhì)暴露的健康人群組成。問卷調(diào)查研究對象的姓名、性別、出生日期、吸煙、飲酒和職業(yè)史等基本情況。2研究方法2.1暴露檢測:對車間空氣中COEs濃度進(jìn)行檢測并估算其累積暴露劑量。采用HPLC法進(jìn)行班末尿中1-羥基芘、3-羥基菲和2-羥基萘濃度測定。2.2效應(yīng)標(biāo)志檢測:采集研究對象外周血,應(yīng)用q PCR的技術(shù)測定外周血白細(xì)胞DNA端粒長度,試劑盒測定血清丙二醛、過氧化氫、過氧化氫酶和總抗氧化能力。2.3基因檢測:采用PCR與限制性片段長度多態(tài)性方法進(jìn)行端粒通路基因、細(xì)胞周期調(diào)控基因和代謝酶基因多態(tài)性檢測。采用q PCR的方法檢測相關(guān)基因m RNA表達(dá)。3統(tǒng)計(jì)學(xué)分析采用SPSS21.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析,非正態(tài)分布數(shù)據(jù)用M(P25,P75)進(jìn)行統(tǒng)計(jì)描述,組間比較用秩和檢驗(yàn),分類變量組間比較采用?2檢驗(yàn)。采用stata13.0統(tǒng)計(jì)軟件對端粒長度的影響因素進(jìn)行多因素分位數(shù)回歸分析。檢驗(yàn)水準(zhǔn)α=0.05。基準(zhǔn)劑量分析采用美國環(huán)境保護(hù)局發(fā)布的BMDS 2.6.0.1軟件。結(jié)果1.焦?fàn)t工職業(yè)暴露調(diào)查暴露COEs時(shí)間加權(quán)平均濃度(Time weighted average concentration,TWA)超標(biāo)的工種主要有裝煤車司機(jī)、攔焦車司機(jī)、焦側(cè)爐門工、上升管工和熱修工。COEs暴露組累積暴露劑量為1.12(0.34,2.14)mg/m3·年,尿1-羥基芘和3-羥基菲濃度分別為83.80(40.29,163.55)pg/μg肌酐和18.20(9.99,35.63)pg/μg肌酐,影響因素為性別、年齡、吸煙、飲酒和工齡,2-羥基萘濃度為84.97(46.49,174.45)pg/μg肌酐,主要影響因素為性別、年齡和吸煙。2.焦?fàn)t工外周血DNA端粒長度和氧化損傷效應(yīng)探討暴露組和對照組端粒長度分別為0.75(0.51,1.08)和1.05(0.76,1.44),兩組差異有統(tǒng)計(jì)學(xué)意義(Z=7.692,P0.001);暴露組與對照組的總抗氧化能力為分別為11.96(8.88,14.68)和14.55(10.51,19.24),差異有統(tǒng)計(jì)學(xué)意義(Z=6.614,P0.001)。氧化損傷各指標(biāo)與端粒長度無明顯相關(guān)性�；鶞�(zhǔn)劑量(Benchmark dose,BMD)研究顯示:外暴露COEs濃度的BMDL值為1.839 mg/m3·年,尿代謝物1-羥基芘BMDL值為43.254(pg/μg肌酐),3-羥基菲BMDL值為11.369(pg/μg肌酐)。3.焦?fàn)t工端粒長度與基因多態(tài)性和相關(guān)m RNA表達(dá)的關(guān)系暴露組CYP2E1基因rs3813867位點(diǎn)CC基因型個體端粒長度為0.97(0.70,1.20),長于GG型個體的端粒長度0.73(0.50,1.09),差異有統(tǒng)計(jì)學(xué)意義(Z=2.536,P=0.011)。暴露組P21、TPP1、TRF1和TRF2基因m RNA表達(dá)低于對照組(P0.05)。暴露組P53和POT1的m RNA表達(dá)高于對照組(P0.05)。P53、POT1、TIN2和TRF2基因m RNA表達(dá)與端粒長度均呈負(fù)相關(guān)(P0.05),TRF1基因m RNA表達(dá)與端粒長度呈正相關(guān)(P=0.015)。COEs暴露組POT1基因rs10250202位點(diǎn)AA基因型個體的POT1基因m RNA表達(dá)低于對照組(P=0.041);COEs暴露組P21基因rs1801270和rs1059234位點(diǎn)的各基因型個體的P21基因m RNA表達(dá)均低于對照組(P0.05)。COEs暴露組P53基因rs1042522、rs1625895位點(diǎn)中的各基因型個體和rs17878362位點(diǎn)SS基因型個體的P53基因m RNA表達(dá)均高于對照組(P0.05);COEs暴露組TRF1基因rs3863242位點(diǎn)中的各基因型個體的TRF1基因m RNA表達(dá)均低于對照組(P0.001)。4.焦?fàn)t工端粒長度影響因素的多因素分析在第15th~85th分位數(shù)下暴露分組因素與端粒長度呈負(fù)相關(guān)。性別因素與第75th分位數(shù)下的端粒長度呈正相關(guān)。年齡與第50th分位數(shù)下的端粒長度呈負(fù)相關(guān)。暴露濃度與第5th~25th分位數(shù)下的端粒長度呈負(fù)相關(guān)。TERT基因rs2736098位點(diǎn)多態(tài)性與第75th分位數(shù)下的端粒長度呈正相關(guān)。P53基因rs17878362位點(diǎn)多態(tài)性與第75th~95th分位數(shù)下的端粒長度呈正相關(guān)。CYP2E1 rs3813867位點(diǎn)多態(tài)性與第25th分位數(shù)下的端粒長度呈正相關(guān)。TPP1基因m RNA表達(dá)量與第75th和85th分位數(shù)下的端粒長度呈正相關(guān)。TRF1基因m RNA表達(dá)量與第75th分位數(shù)下的端粒長度呈負(fù)相關(guān)。P53基因m RNA表達(dá)量與第75th分位數(shù)下的端粒長度呈負(fù)相關(guān)。RAP基因m RNA表達(dá)量與第15th分位數(shù)下的端粒長度呈負(fù)相關(guān)。TIN2基因m RNA表達(dá)量與第85th分位數(shù)下的端粒長度呈負(fù)相關(guān)。結(jié)論1.外周血白細(xì)胞DNA端粒長度和尿代謝物中1-羥基芘可能作為焦?fàn)t工人效應(yīng)生物標(biāo)志和暴露生物標(biāo)志,采用BMD方法獲得的COEs和1-羥基芘的BMDL可為COEs車間空氣濃度限值和職業(yè)接觸生物限值的制定提供依據(jù)。2.單因素分析發(fā)現(xiàn),CYP2E1 rs3813867位點(diǎn)多態(tài)對外周血白細(xì)胞端粒DNA長度有影響,可能是端粒長度的易感性生物標(biāo)志。端粒長度的縮短可能與P53、POT1、TIN2和TRF2 m RNA表達(dá)量增高及TRF 1 m RNA表達(dá)降低有關(guān)。3.分位數(shù)回歸方法探討端粒長度影響因素:暴露濃度,P53、RAP、TIN2和TRF1等基因m RNA的表達(dá)升高可能為端粒長度的危險(xiǎn)因素。TPP1基因m RNA表達(dá)升高,TERT基因rs2736098位點(diǎn)AA基因型、P53基因rs17878362位點(diǎn)的SL基因型、CYP2E1基因rs3813867位點(diǎn)CC基因型可能是端粒長度的保護(hù)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.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R135
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本文編號：1919118