室內(nèi)裝修苯污染對機體的遺傳損傷效應及機制探討
發(fā)布時間：2018-08-06 09:55
【摘要】： 苯(Benzene)是一種工業(yè)用途廣泛的職業(yè)有害因素和室內(nèi)環(huán)境污染物。在工業(yè)中應用于油漆、農(nóng)藥、合成橡膠、染料和皮革加工等行業(yè)。同時苯也是一種主要的室內(nèi)空氣污染物,室內(nèi)空氣中的苯主要來源于油漆、墻面涂料、裝飾材料等。調查顯示:室內(nèi)裝修苯污染可致居住人群健康損害,主要表現(xiàn)為神經(jīng)系統(tǒng)和造血系統(tǒng)損害。目前普遍認為苯是人類致癌和致突變物,高水平的苯接觸也可能引起白血病和不育、流產(chǎn)、畸形、智障等重大遺傳性疾病。如:北京兒童醫(yī)院的統(tǒng)計數(shù)據(jù)表明,該醫(yī)院診治的兒童白血病中80%以上家庭近年進行過新的室內(nèi)裝修。由此可見,繼續(xù)深入開展對苯的遺傳毒性研究是很有意義的,尤其是運用新技術在細胞和分子水平上的研究,將從更深層次揭示苯的遺傳毒性及其作用機理。 目的本研究旨在明確室內(nèi)裝修苯污染狀況,探討苯及其代謝物氫醌對機體的遺傳毒性效應和作用機理,為安全性評價提供實驗及理論依據(jù)。 方法采用國標方法對南寧市近5年新裝修的124戶居室空氣中苯、甲苯、二甲苯濃度進行檢測,并對苯超標戶的家庭成員進行健康問卷調查,觀察苯污染對居住人群身體健康的影響。同時無菌條件采集健康成人外周靜脈血,用常規(guī)離心方法體外分離培養(yǎng)淋巴細胞,設低、中、高濃度苯(0.25、3.5、50.85μmol/L)和氫醌(50、150、450μmol/L)受試物組,另設空白對照組和溶劑對照組。細胞經(jīng)培養(yǎng)處理后應用Annexin V-FITC/PI雙染結合流式細胞術分析人淋巴細胞凋亡情況;采用單細胞凝膠電泳(SCGE)方法和微核實驗方法檢測DNA損傷作用;用衰減全反射紅外光譜技術(ATR)在900～1900 cm-1波段檢測蛋白質、DNA、RNA、糖蛋白和蛋白磷酸化等水平和結構的改變。 結果 1.南寧市近5年新裝修的124戶居室空氣中污染物苯、甲苯、二甲苯的超標率分別為2.4%、10.5%和8.1%,苯超標戶的家庭成員中出現(xiàn)注意力不集中、煩躁、頭暈、易疲勞、易感冒、呼吸道炎癥、胸悶、惡心癥狀,比對照組比照有增多趨向,但無統(tǒng)計學意義。 2.體外實驗中,苯各濃度組(高、中、低)總凋亡率分別為92.85%、74.22%、60.73%;氫醌各濃度組(高、中、低)總凋亡率分別為分別為97.65%、95.09%、54.36%,并可見隨著苯和氫醌染毒劑量的增加,凋亡細胞百分比均逐漸增加,各染毒組細胞凋亡率高于對照組(P0.05或P0.01)。 3.與溶劑對照組比較,苯各濃度組(高、中、低)的彗星率、彗星尾長和微核率分別增加90.9～2527.3%,78.3～118.2%和11.7～31.5% (P0.05或P0.01)。與低濃度組比較,氫醌中和高濃度組彗星率和彗星尾長分別增加13.0～58.9%和213.5～215.1%(P 0.01),與空白對照組比較,氫醌各濃度組的微核率增加400～3076%(P0.01)。 4.苯和氫醌ATR光譜圖呈明顯差異,標識DNA/RNA的光譜區(qū)域(1000～1490cm-1)峰型改變明顯。用主成分統(tǒng)計分析,在三維空間旋轉圖上苯和氫醌點的分布與對照組差異明顯。 結論 1.苯和氫醌均可誘導人淋巴細胞凋亡,提示苯和氫醌可致人淋巴細胞凋亡調節(jié)紊亂,凋亡調節(jié)紊亂有助于細胞的惡性轉化和腫瘤的進展,是惡性腫瘤形成的重要機制之一。 2.高濃度苯和氫醌均可致人淋巴細胞DNA斷裂,具有明顯的遺傳毒性作用, DNA斷裂是細胞凋亡的主要標志,因此可進一步表明苯和氫醌對人淋巴細胞有明顯促凋亡作用。 3.標識DNA/RNA的光譜區(qū)域(1000～1490cm-1)峰型改變明顯,提示苯和氫醌所誘導的細胞內(nèi)化學物質的改變都與細胞遺傳功能密切相關。ATR光譜技術可作為早期細胞損傷的檢測方法。 4、苯遺傳毒性主要作用機理是苯通過其代謝產(chǎn)物產(chǎn)生的具有強氧化性自由基直接或間接氧化損傷DNA或染色體,續(xù)而導致DNA斷裂。
[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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R114
本文編號：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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R114
【引證文獻】
相關期刊論文 前1條
1 孟國忠;季孔庶;;室內(nèi)空氣污染的植物凈化研究概述[J];林業(yè)科技開發(fā);2013年04期
，本文編號：2167371
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