室内装修苯污染对机体的遗传损伤效应及机制探讨
发布时间：2018-08-06 09:55
【摘要】： 苯(Benzene)是一种工业用途广泛的职业有害因素和室内环境污染物。在工业中应用于油漆、农药、合成橡胶、染料和皮革加工等行业。同时苯也是一种主要的室内空气污染物,室内空气中的苯主要来源于油漆、墙面涂料、装饰材料等。调查显示:室内装修苯污染可致居住人群健康损害,主要表现为神经系统和造血系统损害。目前普遍认为苯是人类致癌和致突变物,高水平的苯接触也可能引起白血病和不育、流产、畸形、智障等重大遗传性疾病。如:北京儿童医院的统计数据表明,该医院诊治的儿童白血病中80%以上家庭近年进行过新的室内装修。由此可见,继续深入开展对苯的遗传毒性研究是很有意义的,尤其是运用新技术在细胞和分子水平上的研究,将从更深层次揭示苯的遗传毒性及其作用机理。 目的本研究旨在明确室内装修苯污染状况,探讨苯及其代谢物氢醌对机体的遗传毒性效应和作用机理,为安全性评价提供实验及理论依据。 方法采用国标方法对南宁市近5年新装修的124户居室空气中苯、甲苯、二甲苯浓度进行检测,并对苯超标户的家庭成员进行健康问卷调查,观察苯污染对居住人群身体健康的影响。同时无菌条件采集健康成人外周静脉血,用常规离心方法体外分离培养淋巴细胞,设低、中、高浓度苯(0.25、3.5、50.85μmol/L)和氢醌(50、150、450μmol/L)受试物组,另设空白对照组和溶剂对照组。细胞经培养处理后应用Annexin V-FITC/PI双染结合流式细胞术分析人淋巴细胞凋亡情况;采用单细胞凝胶电泳(SCGE)方法和微核实验方法检测DNA损伤作用;用衰减全反射红外光谱技术(ATR)在900～1900 cm-1波段检测蛋白质、DNA、RNA、糖蛋白和蛋白磷酸化等水平和结构的改变。 结果 1.南宁市近5年新装修的124户居室空气中污染物苯、甲苯、二甲苯的超标率分别为2.4%、10.5%和8.1%,苯超标户的家庭成员中出现注意力不集中、烦躁、头晕、易疲劳、易感冒、呼吸道炎症、胸闷、恶心症状,比对照组比照有增多趋向,但无统计学意义。 2.体外实验中,苯各浓度组(高、中、低)总凋亡率分别为92.85%、74.22%、60.73%;氢醌各浓度组(高、中、低)总凋亡率分别为分别为97.65%、95.09%、54.36%,并可见随着苯和氢醌染毒剂量的增加,凋亡细胞百分比均逐渐增加,各染毒组细胞凋亡率高于对照组(P0.05或P0.01)。 3.与溶剂对照组比较,苯各浓度组(高、中、低)的彗星率、彗星尾长和微核率分别增加90.9～2527.3%,78.3～118.2%和11.7～31.5% (P0.05或P0.01)。与低浓度组比较,氢醌中和高浓度组彗星率和彗星尾长分别增加13.0～58.9%和213.5～215.1%(P 0.01),与空白对照组比较,氢醌各浓度组的微核率增加400～3076%(P0.01)。 4.苯和氢醌ATR光谱图呈明显差异,标识DNA/RNA的光谱区域(1000～1490cm-1)峰型改变明显。用主成分统计分析,在三维空间旋转图上苯和氢醌点的分布与对照组差异明显。 结论 1.苯和氢醌均可诱导人淋巴细胞凋亡,提示苯和氢醌可致人淋巴细胞凋亡调节紊乱,凋亡调节紊乱有助于细胞的恶性转化和肿瘤的进展,是恶性肿瘤形成的重要机制之一。 2.高浓度苯和氢醌均可致人淋巴细胞DNA断裂,具有明显的遗传毒性作用, DNA断裂是细胞凋亡的主要标志,因此可进一步表明苯和氢醌对人淋巴细胞有明显促凋亡作用。 3.标识DNA/RNA的光谱区域(1000～1490cm-1)峰型改变明显,提示苯和氢醌所诱导的细胞内化学物质的改变都与细胞遗传功能密切相关。ATR光谱技术可作为早期细胞损伤的检测方法。 4、苯遗传毒性主要作用机理是苯通过其代谢产物产生的具有强氧化性自由基直接或间接氧化损伤DNA或染色体,续而导致DNA断裂。
[Abstract]:Benzene (Benzene) is a widely used occupational hazard and indoor environmental pollutant. It is used in industries such as paints, pesticides, synthetic rubber, dyestuff and leather processing. Benzene is also a major indoor air pollutant. The benzene in indoor air is mainly derived from paint, wall coating, decoration materials and so on. It is generally believed that benzene is human carcinogenic and mutagenesis, and the high level of benzene exposure may also cause major genetic diseases such as leukemia and infertility, abortion, malformation and mental retardation. For example, statistical data sheet of Beijing Children's Hospital In recent years, more than 80% of the families of children with leukemia treated in this hospital have undergone new indoor decoration. Thus, it is significant to continue to carry out the research on the genetic toxicity of benzene, especially with the use of new technologies at the cellular and molecular levels to reveal the genetic toxicity and mechanism of benzene from a deeper level.
Objective to clarify the status of benzene pollution in indoor decoration and explore the effect of benzene and its metabolite hydroquinone on the organism's genotoxic effect and mechanism, so as to provide experimental and theoretical basis for safety evaluation.
Methods the national standard method was used to detect the concentration of benzene, toluene and xylene in the air of 124 newly decorated rooms in Nanning city in recent 5 years. The health questionnaire of the family members of the households with benzene exceeding the standard was investigated to observe the effect of benzene pollution on the health of the living people. Lymphocytes were isolated and cultured in vitro, and a group of low, medium and high concentrations of benzene (0.25,3.5,50.85 mu mol/L) and hydroquinone (50150450 u mol/L) were set up, and a blank control group and a solvent control group were set up. The cells were treated with Annexin V-FITC/PI double dye binding flow cytometry to analyze the apoptosis of human lymphocytes. Single cell gel electrophoresis (SCGE) was used. Methods and micronucleus methods were used to detect the effect of DNA damage, and the level and structure of protein, DNA, RNA, glycoprotein and protein phosphorylation were detected by attenuated total reflectance infrared spectroscopy (ATR) in the 900~1900 cm-1 band.
Result
1. over the past 5 years in Nanning, the excess rates of benzene, toluene and xylene were 2.4%, 10.5% and 8.1% of the pollutants in the 124 newly decorated rooms, and the families of benzene over standard households were not focused, irritable, dizzy, tired, susceptible to cold, respiratory inflammation, chest tightness, and nausea, but there was no statistical significance in comparison with the control group.
2. in vitro, the total apoptosis rates of each group (high, medium and low) were 92.85%, 74.22%, 60.73%, respectively, and the total apoptosis rates of hydroquinone concentration groups (high, medium and low) were 97.65%, 95.09%, 54.36% respectively, and the percentage of dead cells increased gradually with the increase of the dose of benzene and hydroquinone, and the apoptosis rate of each group was higher than that of the control group. P0.05 or P0.01).
3. compared with the solvent control group, the comet rate of each concentration group (high, medium and low), the tail length and the micronucleus rate of the comets increased by 90.9 to 2527.3%, 78.3 to 118.2% and 11.7 to 31.5% (P0.05 or P0.01). Compared with the low concentration group, the comet rate and the comet tail length of the hydroquinone neutralization group increased 13 to 58.9% and 213.5 ~ 215.1% (P 0.01), respectively. The micronucleus rate of each group of hydroquinone increased by 400 to 3076% (P0.01).
The ATR spectra of 4. benzene and hydroquinone were distinctly different, and the peak pattern of the spectral region (1000 ~ 1490cm-1) marked DNA/RNA was obvious. The distribution of benzene and hydroquinone points on the three-dimensional space rotation map was significantly different from that of the control group.
conclusion
1. benzene and hydroquinone can induce apoptosis of human lymphocytes. It is suggested that benzene and hydroquinone can regulate the disorder of human lymphocyte apoptosis, and the disorder of apoptosis regulation contributes to the malignant transformation of cells and the progression of tumor. It is one of the important mechanisms of malignant tumor formation.
2. high concentration of benzene and hydroquinone can break the DNA of human lymphocytes and have obvious genetic toxicity. DNA fracture is the main marker of cell apoptosis. Therefore, it can be further indicated that benzene and hydroquinone have obvious effect on the apoptosis of human lymphocytes.
3. identification of the spectral region (1000 ~ 1490cm-1) of the DNA/RNA is obvious. It is suggested that the changes in the intracellular chemicals induced by benzene and hydroquinone are closely related to the cell genetic function..ATR spectroscopy can be used as a detection method for early cell damage.
4, the main mechanism of genetic toxicity of benzene is that benzene has a strong oxidative free radical produced by its metabolites, directly or indirectly, to directly or indirectly oxidize DNA or chromosomes, which causes DNA to break.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R114
本文编号：2167371
[Abstract]:Benzene (Benzene) is a widely used occupational hazard and indoor environmental pollutant. It is used in industries such as paints, pesticides, synthetic rubber, dyestuff and leather processing. Benzene is also a major indoor air pollutant. The benzene in indoor air is mainly derived from paint, wall coating, decoration materials and so on. It is generally believed that benzene is human carcinogenic and mutagenesis, and the high level of benzene exposure may also cause major genetic diseases such as leukemia and infertility, abortion, malformation and mental retardation. For example, statistical data sheet of Beijing Children's Hospital In recent years, more than 80% of the families of children with leukemia treated in this hospital have undergone new indoor decoration. Thus, it is significant to continue to carry out the research on the genetic toxicity of benzene, especially with the use of new technologies at the cellular and molecular levels to reveal the genetic toxicity and mechanism of benzene from a deeper level.
Objective to clarify the status of benzene pollution in indoor decoration and explore the effect of benzene and its metabolite hydroquinone on the organism's genotoxic effect and mechanism, so as to provide experimental and theoretical basis for safety evaluation.
Methods the national standard method was used to detect the concentration of benzene, toluene and xylene in the air of 124 newly decorated rooms in Nanning city in recent 5 years. The health questionnaire of the family members of the households with benzene exceeding the standard was investigated to observe the effect of benzene pollution on the health of the living people. Lymphocytes were isolated and cultured in vitro, and a group of low, medium and high concentrations of benzene (0.25,3.5,50.85 mu mol/L) and hydroquinone (50150450 u mol/L) were set up, and a blank control group and a solvent control group were set up. The cells were treated with Annexin V-FITC/PI double dye binding flow cytometry to analyze the apoptosis of human lymphocytes. Single cell gel electrophoresis (SCGE) was used. Methods and micronucleus methods were used to detect the effect of DNA damage, and the level and structure of protein, DNA, RNA, glycoprotein and protein phosphorylation were detected by attenuated total reflectance infrared spectroscopy (ATR) in the 900~1900 cm-1 band.
Result
1. over the past 5 years in Nanning, the excess rates of benzene, toluene and xylene were 2.4%, 10.5% and 8.1% of the pollutants in the 124 newly decorated rooms, and the families of benzene over standard households were not focused, irritable, dizzy, tired, susceptible to cold, respiratory inflammation, chest tightness, and nausea, but there was no statistical significance in comparison with the control group.
2. in vitro, the total apoptosis rates of each group (high, medium and low) were 92.85%, 74.22%, 60.73%, respectively, and the total apoptosis rates of hydroquinone concentration groups (high, medium and low) were 97.65%, 95.09%, 54.36% respectively, and the percentage of dead cells increased gradually with the increase of the dose of benzene and hydroquinone, and the apoptosis rate of each group was higher than that of the control group. P0.05 or P0.01).
3. compared with the solvent control group, the comet rate of each concentration group (high, medium and low), the tail length and the micronucleus rate of the comets increased by 90.9 to 2527.3%, 78.3 to 118.2% and 11.7 to 31.5% (P0.05 or P0.01). Compared with the low concentration group, the comet rate and the comet tail length of the hydroquinone neutralization group increased 13 to 58.9% and 213.5 ~ 215.1% (P 0.01), respectively. The micronucleus rate of each group of hydroquinone increased by 400 to 3076% (P0.01).
The ATR spectra of 4. benzene and hydroquinone were distinctly different, and the peak pattern of the spectral region (1000 ~ 1490cm-1) marked DNA/RNA was obvious. The distribution of benzene and hydroquinone points on the three-dimensional space rotation map was significantly different from that of the control group.
conclusion
1. benzene and hydroquinone can induce apoptosis of human lymphocytes. It is suggested that benzene and hydroquinone can regulate the disorder of human lymphocyte apoptosis, and the disorder of apoptosis regulation contributes to the malignant transformation of cells and the progression of tumor. It is one of the important mechanisms of malignant tumor formation.
2. high concentration of benzene and hydroquinone can break the DNA of human lymphocytes and have obvious genetic toxicity. DNA fracture is the main marker of cell apoptosis. Therefore, it can be further indicated that benzene and hydroquinone have obvious effect on the apoptosis of human lymphocytes.
3. identification of the spectral region (1000 ~ 1490cm-1) of the DNA/RNA is obvious. It is suggested that the changes in the intracellular chemicals induced by benzene and hydroquinone are closely related to the cell genetic function..ATR spectroscopy can be used as a detection method for early cell damage.
4, the main mechanism of genetic toxicity of benzene is that benzene has a strong oxidative free radical produced by its metabolites, directly or indirectly, to directly or indirectly oxidize DNA or chromosomes, which causes DNA to break.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R114
【引证文献】
相关期刊论文 前1条
1 孟国忠;季孔庶;;室内空气污染的植物净化研究概述[J];林业科技开发;2013年04期
，本文编号：2167371
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