铅致人淋巴细胞DNA双链断裂作用的流行病学调查和体外实验研究

发布时间：2018-05-06 04:29

本文选题：铅 + 淋巴细胞　；参考：《浙江大学》2012年硕士论文

【摘要】：目的应用流式细胞术检测γH2AX蛋白,结合人群调查和体外实验,分析铅致人淋巴细胞DNA双链断裂作用,探索流式细胞术检测γH2AX以评价人群DNA双链断裂水平的可行性。方法选取某蓄电池厂工人67例和对照人群70例,采外周静脉血提取淋巴细胞,利用流式细胞术检测γH2AX蛋白,分析淋巴细胞DNA双链断裂水平；不同剂量、时间下醋酸铅染毒健康人外周血淋巴细胞,利用流式细胞术检测γH2AX蛋白,分析淋巴细胞DNA双链断裂水平。结果人群调查结果显示高浓度组(41.76%±28.57%；9.90±3.35)、低浓度组(33.18%±30.64%；9.39±4.83)、内对照组(35.87%±34.09%；10.04±5.77)DNA损伤率和平均荧光强度均高于外对照组(0.28%±0.28%；6.95±2.93),差异有统计学意义(P0.05)；性别、吸烟、饮酒、工龄对职业铅暴露工人DNA损伤水平无影响(P0.05)；偏相关分析结果显示内对照组DNA损伤率和平均荧光强度与年龄存在相关性(r=-0.430、-0.391,P0.05),铅暴露高浓度组平均荧光强度与血铅、δ-ALA存在相关性(r=0.621、-0.697, P0.05)。体外实验结果显示1h和2h染毒组中除62.5μmol/L外,125μmol/L、250μmol/L、500μmol/LDNA损伤率均与阴性对照组、阳性对照组存在统计学差异(P0.01),随着染毒剂量增高,DNA损伤率呈现先增高后降低趋势；各浓度组吸光度值均低于阴性对照组,但未见剂量效应关系,2h组吸光度值均低于1h组。结论铅致DNA双链断裂损伤可能是DNA修复抑制、DNA断裂、DNA-DNA交联、DNA-蛋白质交联多方面作用因素共同结果,流式细胞术检测yH2AX是一种值得运用于检测大样本DNA双链断裂水平的方法。
[Abstract]:Objective to detect 纬 H2AX protein by flow cytometry, analyze the double strand breaks of DNA in human lymphocytes induced by lead, and explore the feasibility of detecting 纬 H2AX by flow cytometry to evaluate the level of DNA double strand breaks in human lymphocytes. Methods 67 workers from a storage battery factory and 70 controls were selected to extract lymphocytes from peripheral venous blood. Flow cytometry was used to detect 纬 H2AX protein and analyze the level of DNA double strand breaks in lymphocytes. The 纬 H2AX protein was detected by flow cytometry and the level of DNA double strand break in lymphocytes was analyzed by flow cytometry. Results the results of population investigation showed that the damage rate and average fluorescence intensity of 5.77)DNA in high concentration group (41.76% 卤28.576.95 卤3.35), low concentration group (33.18% 卤30.64 卤9.39 卤4.83) and internal control group (35.87% 卤34.090.04 卤10.04 卤2.93g) were higher than those in external control group (0.28% 卤0.286.95 卤2.93g), the difference was significant (P 0.05). The results of partial correlation analysis showed that there was a correlation between the DNA damage rate and average fluorescence intensity and age in the control group, the average fluorescence intensity in the lead exposure group was correlated with the blood lead, the 未 -ALA level was correlated with r0.621-0.697, P0.05. The results of in vitro experiments showed that the damage rate of 125渭 mol / L ~ (125) 渭 mol / L ~ (250 渭 mol / L) ~ (500 渭 mol / L) mol/LDNA in 1 h and 2 h exposure group was significantly higher than that in the negative control group (P < 0.01), and the damage rate increased firstly and then decreased with the increase of dose. The absorbance value of each concentration group was lower than that of negative control group, but the absorbance value of 2 h group was lower than that of 1 h group. Conclusion DNA double strand break damage induced by lead may be the result of DNA repair inhibiting DNA-DNA cross-linking DNA-protein crosslinking. Flow cytometry is a valuable method to detect the level of DNA double strand breaks in large samples.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R181.3

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