發(fā)布時(shí)間：2018-04-15 21:39

  本文選題：鋁 + 認(rèn)知功能��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:本次研究采用分子流行病學(xué)方法在職業(yè)性鋁接觸人群中開展研究,評(píng)估長(zhǎng)期職業(yè)性鋁接觸人群認(rèn)知功能改變情況,分析血鋁與外周血淋巴細(xì)胞H3K4me3、H3K9me2、H3K27me3及血漿BDNF、EGR1蛋白水平之間的關(guān)系,初步探討長(zhǎng)期職業(yè)性鋁接觸引起的組蛋白賴氨酸甲基化譜改變情況,為長(zhǎng)期鋁暴露的分子流行病學(xué)研究提供參考依據(jù),并為以后研究上述指標(biāo)是否可作為從外周血水平對(duì)鋁的神經(jīng)毒性進(jìn)行監(jiān)測(cè)和早期診斷的替代生物標(biāo)志物提供人群分子流行病學(xué)依據(jù)。方法:采用整群抽樣的方式,選擇山西某鋁廠電解鋁車間100例男性工人作為暴露組、熱電車間135例男性工人作為對(duì)照組。采用自行設(shè)計(jì)的職業(yè)流行病學(xué)調(diào)查問卷:簡(jiǎn)短精神狀態(tài)量表(MMSE)、數(shù)字廣度測(cè)試[DST,包括正序測(cè)試(DSFT)和倒序測(cè)試(DSBT)]、畫鐘試驗(yàn)(CDT)、物體記憶測(cè)驗(yàn)(FOME)、言語流暢性測(cè)驗(yàn)(VFT)等進(jìn)行調(diào)查,收集研究對(duì)象的一般情況并進(jìn)行認(rèn)知功能評(píng)分;采集調(diào)查對(duì)象的空腹肘靜脈血液,分離血漿并提取淋巴細(xì)胞,-80℃凍存?zhèn)溆?采用石墨爐原子吸收法測(cè)定血鋁含量,可能由于該大型鋁廠對(duì)當(dāng)?shù)丨h(huán)境的影響,對(duì)照組和暴露組工人的血鋁水平無統(tǒng)計(jì)學(xué)差異,故以血鋁水平的上下百分位數(shù)P25、P75和中位數(shù)M將研究對(duì)象分為低、中、高血鋁組;酶聯(lián)免疫吸附試驗(yàn)(ELISA)方法定量檢測(cè)淋巴細(xì)胞中H3K4me3、H3K9me2、H3K27me3表達(dá)水平和血漿BDNF、EGR1蛋白水平。結(jié)果:1.職業(yè)性鋁接觸工人認(rèn)知功能與血漿BDNF、EGR1的關(guān)系:(1)職業(yè)性鋁接觸工人的血鋁水平比較:接觸組工人血鋁值為(155.07±71.17)μg/L,對(duì)照組工人血鋁值為(142.94±53.80)μg/L,兩組差異無統(tǒng)計(jì)學(xué)意義(F=1.488,P0.05)。下四分位數(shù)P25為100.14mg/L、中位數(shù)M為134.36mg/L、上四分位數(shù)P75為178.96mg/L,故按照血鋁水平的四分位數(shù):m(p25,p75)為134.36(100.14,178.96)mg/l,將所有研究對(duì)象分為低血鋁組82.88(72.69,92.40)mg/l、中血鋁組134.46(115.74,154.68)mg/l和高血鋁組228.59(204.91,263.87)mg/l。(2)不同血鋁水平工人的基本情況比較:不同血鋁水平工人在年齡、工齡、受教育程度及吸煙、飲酒方面,差異均無統(tǒng)計(jì)學(xué)意義(均p0.05)。(3)不同血鋁水平工人的認(rèn)知功能評(píng)價(jià):不同血鋁水平工人的mmse、dsft、dsbt、dst得分不同,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05)。高血鋁組工人mmse、dsft、dsbt、dst的平均得分均分別低于低血鋁組工人的平均得分,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05);中血鋁組工人mmse、dsbt、dst的平均得分均分別低于低血鋁組工人的平均得分,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05)。(4)血鋁與認(rèn)知功能的相關(guān)分析結(jié)果顯示:血鋁水平和認(rèn)知功能之間具有不同程度的相關(guān)性。血鋁水平與mmse評(píng)分之間存在負(fù)相關(guān)(rs=-0.252,p0.01),與dsft評(píng)分之間存在負(fù)相關(guān)(rs=-0.197,p0.01),與dst評(píng)分之間存在負(fù)相關(guān)性(rs=-0.181,p0.01),說明隨著血鋁水平的升高,認(rèn)知功能評(píng)分中的mmse、dsft、dst得分均呈降低趨勢(shì)。(5)影響認(rèn)知功能的多因素回歸分析結(jié)果顯示;血鋁、年齡、工齡、受教育程度分別被納入回歸方程,差異有統(tǒng)計(jì)學(xué)意義。說明年齡、工齡、受教育程度均可能影響認(rèn)知功能。(6)工人血漿bdnf、egr1蛋白水平分層分析結(jié)果顯示:不同血鋁組工人血漿bdnf蛋白(f=5.385,p=0.005)表達(dá)量、egr1蛋白(f=6.853,p=0.001)表達(dá)量不同,差異有統(tǒng)計(jì)學(xué)意義。與低血鋁組相比,高、中血鋁組工人血漿bdnf表達(dá)量均分別降低,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05)。高血鋁組工人血漿egr1蛋白表達(dá)量分別低于低、中血鋁組工人血漿egr1蛋白表達(dá)量,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05)。但是按照年齡、工齡、受教育程度分層分析后,結(jié)果均顯示:血漿bdnf、egr1蛋白水平在各組間表達(dá)量差異并無統(tǒng)計(jì)學(xué)意義。(7)血鋁、bdnf、egr1蛋白及認(rèn)知功能的相關(guān)性分析結(jié)果:血鋁水平、bdnf、egr1和認(rèn)知功能之間具有不同程度的相關(guān)性。血鋁水平與bdnf蛋白、egr1蛋白水平之間均存在負(fù)相關(guān)性(r=-0.214,p0.01、r=-0.315,p0.01),說明隨著血鋁水平的升高,bdnf和egr1的表達(dá)量均呈下降趨勢(shì);bdnf蛋白水平與mmse評(píng)分之間存在正相關(guān)性(rs=0.140,p0.05),egr1蛋白水平與mmse評(píng)分之間存在正相關(guān)性(rs=0.201,p0.05),說明隨著bdnf、egr1的表達(dá)量降低,mmse評(píng)分呈降低趨勢(shì)。2.職業(yè)性鋁接觸工人組蛋白賴氨酸甲基化修飾改變及影響分析:(1)不同血鋁水平工人外周血淋巴細(xì)胞h3k4me3水平比較:不同血鋁水平工人外周血淋巴細(xì)胞h3k4me3的表達(dá)量不同,差異有統(tǒng)計(jì)學(xué)意義(f=3.572,p=0.031)。高血鋁組工人h3k4me3表達(dá)量低于低血鋁組h3k4me3表達(dá)量,差異有統(tǒng)計(jì)學(xué)意義(p0.05);高血鋁組h3k4me3表達(dá)量低于中血鋁組h3k4me3表達(dá)量,中血鋁組h3k4me3表達(dá)量低于低血鋁組h3k4me3表達(dá)量,但差異均無統(tǒng)計(jì)學(xué)意義(均p0.05)。(2)不同血鋁水平工人外周血淋巴細(xì)胞h3k9me2水平比較:不同血鋁水平工人外周血淋巴細(xì)胞h3k9me2的表達(dá)量不同,差異有統(tǒng)計(jì)學(xué)意義(f=9.551,p=0.001)。高血鋁組工人h3k9me2表達(dá)量高于低血鋁組和中血鋁組工人h3k9me2的表達(dá)量,差異均有統(tǒng)計(jì)學(xué)意義(均p0.05);中血鋁組工人h3k9me2表達(dá)量高于低血鋁組工人h3k4me3表達(dá)量,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。(3)不同血鋁水平工人外周血淋巴細(xì)胞h3k27me3水平比較:不同血鋁水平工人外周血淋巴細(xì)胞h3k27me3的表達(dá)量不同,差異有統(tǒng)計(jì)學(xué)意義(f=3.641,p=0.028)。高血鋁組工人h3k27me3的表達(dá)量高于低血鋁組工人h3k27me3的表達(dá)量,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。(4)淋巴細(xì)胞h3k4me3、h3k9me2、h3k27me3與血鋁的相關(guān)性分析:h3k4me3、h3k9me2、h3k27me3表達(dá)量和血鋁之間存在不同程度的相關(guān)性。其中h3k4me3和血鋁之間呈負(fù)相關(guān)(r=-0.307,p0.01),說明隨著血鋁水平的升高,職業(yè)性鋁接觸工人外周血淋巴細(xì)胞h3k4me3表達(dá)量降低;h3k9me2、h3k27me3與血鋁之間均呈正相關(guān)(r=0.329、r=0.387,均p0.01),說明隨著血鋁水平升高,職業(yè)性鋁接觸工人外周血淋巴細(xì)胞h3k9me2、h3k27me3表達(dá)量也升高。(5)bdnf、egr1和h3k4me3、h3k9me2、h3k27me3的相關(guān)分析結(jié)果:bdnf、egr1蛋白水平和h3k4me3、h3k9me2、h3k27me3表達(dá)量之間存在不同程度的相關(guān)性。其中bdnf、egr1和h3k4me3之間均呈正相關(guān)(r=0.202,p0.05、r=0.361,p0.01),說明隨著h3k4me3表達(dá)量的降低,bdnf、egr1兩種蛋白的表達(dá)量也降低;bdnf和h3k9me2之間呈負(fù)相關(guān)(r=-0.286,p0.01),說明隨著h3k9me2表達(dá)量升高,bdnf的表達(dá)量降低;egr1和h3k27me3之間呈負(fù)相關(guān)(r=-0.241,p0.05),說明隨著h3k27me3表達(dá)量升高,egr1蛋白的表達(dá)量降低。(6)血漿bdnf、egr1蛋白影響因子的多元逐步回歸分析:bdnf與h3k4me3、h3k9me2之間存在線性回歸關(guān)系,h3k9me2對(duì)bdnf的影響比h3k4me3對(duì)bdnf的影響大;egr1與h3k4me3、h3k27me3之間存在線性回歸關(guān)系,h3k4me3對(duì)egr1的影響比h3k27me3對(duì)egr1的影響大。結(jié)論:1.長(zhǎng)期職業(yè)性鋁接觸可引起工人認(rèn)知功能改變和血漿BDNF、EGR1水平降低,且血鋁、認(rèn)知功能和BDNF、EGR1之間存在一定的相關(guān)性,提示鋁可能通過影響B(tài)DNF、EGR1的表達(dá)影響認(rèn)知功能。2.血鋁水平、BDNF、EGR1和H3K4me3、H3K9me2、H3K27me3的表達(dá)水平之間均有一定的相關(guān)性,表明在職業(yè)性鋁接觸引起的認(rèn)知功能改變過程中,鋁可能通過下調(diào)H3K4me3、上調(diào)H3K27me3及H3K9me2、下調(diào)記憶相關(guān)蛋白BDNF和EGR1的表達(dá)引起認(rèn)知功能改變。
[Abstract]:Objective: This study adopts the methods of molecular epidemiology research in occupation of aluminum exposed, the change of occupation of aluminum exposed population to assess the long-term cognitive function, analysis of blood aluminum and peripheral blood lymphocyte H3K4me3, H3K9me2, H3K27me3 and BDNF in plasma, the relationship between EGR1 protein level and preliminary study of aluminum exposure caused by long-term occupation histone lysine methylation profiles, provide a reference basis for the molecular epidemiology of long-term exposure to aluminum, and to investigate whether these indexes can provide the basis for molecular epidemiology as surrogate biomarkers from the peripheral blood level of neurotoxicity of aluminum for monitoring and early diagnosis. Methods: cluster the way of sampling, choose a Shanxi aluminum electrolytic aluminum workshop of 100 male workers as exposure group, 135 cases of male power workshop workers as control group. The self set. Occupation questionnaire meter: Mini Mental State Scale (MMSE), digital span test [DST, including positive test (DSFT) and reverse (DSBT) test, clock drawing test (CDT), object memory test (FOME), verbal fluency test (VFT) for investigation, cognitive function the score generally collected and venous blood collection; investigation, separation and extraction of plasma cells, -80 C cryopreserved blood; Determination of aluminium content by graphite furnace atomic absorption spectrometry may be due to the effect of the large aluminum in the environment, no significant difference between the control group and the group of workers exposed to blood aluminum level, so the blood aluminum level on the percentile of P25, P75 and M of the median object is divided into low, high blood aluminum group; enzyme linked immunosorbent assay (ELISA) method for quantitative detection of H3K4me3 in H3K9me2 cells, the expression level of H3K27me3 and blood Plasma BDNF, EGR1 protein level. Results: 1. of workers exposed to aluminum occupation cognitive function and plasma BDNF, EGR1: (1) comparison of blood aluminum level occupation of aluminum exposed workers: exposed workers blood aluminum value (155.07 + 71.17) g/L, group of workers blood aluminum value as control (142.94. 53.80) g/L, with no significant difference between two groups (F=1.488, P0.05). Four digit P25 100.14mg/L, median M 134.36mg/L, four digit P75 178.96mg/L, so in accordance with the four percentile of blood aluminum level: m (P25, p75) 134.36 (100.14178.96) mg/l, all of the subjects were divided into low blood aluminum group 82.88 (72.69,92.40) mg/l in blood aluminum group 134.46 (115.74154.68) mg/l and high blood aluminum group 228.59 (204.91263.87) mg/l. (2) compared to the basic situation of workers of different blood aluminum level: Blood aluminum level in workers of different age, seniority, education, and smoking, drinking. There were no significant differences ( P0.05). (3) to evaluate the cognitive function of workers of different blood aluminum level: workers in different blood aluminum level of MMSE, DSFT, dsbt, DST scores, the differences were statistically significant (P0.05). High blood aluminum group DSFT, dsbt, MMSE workers, the average score of DST were significantly lower than those of the average score of low blood aluminum group of workers, the differences were statistically significant (P0.05); blood group dsbt, MMSE aluminum workers, the average score of DST were significantly lower than those of the average score of low blood aluminum group of workers, the differences were statistically significant (P0.05). (4) the correlation analysis and cognitive function of aluminum. The result showed that the correlation with blood between different levels of functional and cognitive level of aluminum blood. There was a negative correlation between blood aluminum level and MMSE score (rs=-0.252, P0.01), and there was a negative correlation between DSFT scores (rs=-0.197, P0.01), and there is a negative correlation between DST scores (rs=-0.181, P0.01), with elevated blood levels of aluminum,. Know the function score in MMSE, DSFT, DST score decreased. (5) regression analysis showed that the influencing factors of cognitive function; blood aluminum, age, seniority, education level were included in the regression equation, the difference was statistically significant. The age, seniority, education level can affect cognitive function (6) workers. Plasma levels of BDNF, EGR1 protein level stratified analysis showed that different groups of workers blood aluminum plasma BDNF protein (f=5.385, p=0.005) expression of EGR1 protein (f=6.853, p=0.001) expression is different, the difference was statistically significant. Compared with low blood aluminum, high blood aluminum plasma BDNF group of workers the expression amount was decreased, the differences were statistically significant (P0.05). High blood aluminum group workers plasma EGR1 protein expression were lower in blood plasma, aluminum group of workers the expression of EGR1 protein, the differences were statistically significant (all P0.05). But according to the age, length of service, by Analysis of education level stratification, results showed that plasma BDNF expression was no significant difference in EGR1 protein levels in each group. (7) blood aluminum, BDNF, correlation between EGR1 protein and cognitive function analysis results: Blood aluminum level, BDNF, has a different degree of correlation between EGR1 and blood aluminum level of cognitive function. With the BDNF protein, there was a negative correlation between EGR1 protein level (r=-0.214, P0.01, r=-0.315, P0.01), with elevated blood aluminum level, expression of BDNF and EGR1 showed a downward trend; there is a positive correlation between BDNF protein level and MMSE score (rs=0.140, P0.05), there is a positive correlation between EGR1 protein level with the score of MMSE (rs=0.201, P0.05), with BDNF, the expression of EGR1 was decreased and the MMSE score decreased.2. occupation aluminum workers exposed to histone lysine methylation modification and analysis: (1) effects of different blood levels of aluminum industry Comparison of h3k4me3 in human peripheral blood lymphocytes of peripheral blood lymphocyte h3k4me3 expression level: workers with different amount of blood aluminum level is different, the difference was statistically significant (f=3.572, p=0.031). The high expression of h3k4me3 blood group of aluminum workers is lower than the low expression of h3k4me3 in blood aluminum group, the difference was statistically significant (P0.05); high blood aluminum group the expression of h3k4me3 was lower than that in blood group h3k4me3 expression in aluminum, aluminum blood group h3k4me3 expressed lower than low blood aluminum group the expression of h3k4me3, but the differences were not statistically significant (P0.05). (2) comparison of workers of different blood aluminum level H3K9me2 levels in peripheral blood lymphocytes of peripheral blood lymphocytes: the expression of H3K9me2 in different blood aluminum workers the amount of different levels, the difference was statistically significant (f=9.551, p=0.001). The high blood aluminum workers H3K9me2 expression group was higher than that of low blood aluminum group and blood aluminum workers H3K9me2 group, the differences were statistically significant (P0.05); blood Aluminum Group The workers H3K9me2 was higher than that of low blood aluminum group of workers h3k4me3 expression, the difference was statistically significant (P0.05). (3) comparison of workers of different blood aluminum level H3K27me3 levels in peripheral blood lymphocytes of peripheral blood lymphocyte H3K27me3 expression: workers with different amount of blood aluminum level is different, the difference was statistically significant (f=3.641, p=0.028). Expression of high blood aluminum group of workers is higher than the H3K27me3 H3K27me3 aluminum low blood group of workers, the difference was statistically significant (P0.05). (4) h3k4me3 lymphocytes, H3K9me2, correlation analysis of H3K27me3 and blood aluminum: h3k4me3, H3K9me2, there are different degrees of correlation between blood volume and the expression of H3K27me3 was negatively related to aluminum. Between the h3k4me3 and blood aluminum (r=-0.307, P0.01), with elevated blood aluminum level, occupation of aluminum exposed workers peripheral blood lymphocyte h3k4me3 expression decreased; H3K9me2, a positive correlation between H3K27me3 and blood aluminum (r=0. 329, r=0.387, P0.01), with blood aluminum level increased, H3K9me2 in peripheral blood lymphocytes of workers exposed to aluminum occupation, the expression of H3K27me3 was also increased. (5) BDNF, EGR1 and h3k4me3, H3K9me2, correlation analysis results of H3K27me3: BDNF, EGR1 protein level and h3k4me3, H3K9me2, there are different degrees of correlation between the expression of H3K27me3. BDNF, EGR1 and h3k4me3 were positively correlated (r=0.202, P0.05, r=0.361, P0.01, h3k4me3) with reduced expression of BDNF, EGR1, the expression of two proteins decreased; there was a negative correlation between BDNF and H3K9me2 (r=-0.286, P0.01), with the expression of H3K9me2 increased, the expression of BDNF was decreased; there was a negative correlation between EGR1 and H3K27me3 (r=-0.241, P0.05), with H3K27me3 expression increased, EGR1 protein expression decreased. (6) plasma BDNF, stepwise regression analysis of EGR1 protein factors: BDNF and h3k4me3, h3k yuan There is a linear regression relationship between 9me2, the effect of H3K9me2 on the BDNF than the effect of h3k4me3 on BDNF; EGR1 and h3k4me3, there was a linear regression relationship between H3K27me3 and the effect of h3k4me3 on the EGR1 than the effect of H3K27me3 on EGR1. Conclusion: 1. long-term occupation of aluminum exposure may induce changes in cognitive function and plasma BDNF, EGR1 level reduced, and blood aluminum, cognitive function and BDNF, there is a certain correlation between EGR1, suggesting that aluminum may influence the BDNF, the expression of EGR1 on cognitive function in.2. blood aluminum level, BDNF, EGR1 and H3K4me3, H3K9me2, have a certain degree of correlation between the expression level of H3K27me3, showed that cognitive function caused by occupation of aluminum contact the change in the process of aluminum through down-regulation of H3K4me3 and up regulation of H3K27me3 and H3K9me2, downregulation of memory related proteins BDNF and EGR1 caused changes in cognitive function.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R135

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相關(guān)期刊論文 前10條

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