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目的利用细胞受体转录激活试验,研究居室内灰尘有机抽提物的甲状腺激素干扰活性,并分析其中可能的化学物质组成。方法利用HEK-293细胞,将重组报告基因质粒p UAS-tk-Luc、表达质粒Gal4-L-TRβ和Sofast TM转染试剂转染到细胞中。灰尘有机提取物用培养基稀释成不同浓度后与5 nmol/L三碘甲状腺原氨酸(triiodothyronin 3,T3)一起加入细胞培养板,利用荧光素酶报告基因检测仪检测荧光素酶的表达量。同时,测定5类可能影响体内甲状腺激素平衡的有机化合物。结果随着受试液浓度的升高,荧光素酶的表达量降低,细胞内T3的活性降低,线性趋势检验P<0.001,10.00和20.00 mg灰尘/ml浓度组荧光素酶表达量均低于5 nmol/L T3阳性对照组,P分别为0.012,0.002。化学分析显示居室内灰尘中常见内分泌干扰物质含量分别为:多环芳烃类0.2~3.7μg/g,邻苯二甲酸酯类未检出(not detected,ND)~117.4μg/g,多溴联苯醚类ND~1.0μg/g,多氯联苯类1.0~25.0 ng/g,酚类ND~32.5μg/g。结论室内灰尘有机抽提物能够干扰甲状腺激素活性,提示儿童早期室内灰尘暴露对儿童神经发育有潜在的影响。
OBJECTIVE: To study the thyroid hormone interference activity of organic extracts from indoor dust using the cell receptor transcriptional activation assay and to analyze the possible chemical constituents. Methods HEK-293 cells were transfected with recombinant reporter plasmid pUAS-tk-Luc, expression plasmid Gal4-L-TRβ and Sofast TM transfection reagent. Dust organic extracts were diluted into different concentrations with the medium and then added to the cell culture plates together with 5 nmol / L triiodothyronin 3 (T3). The luciferase reporter gene was used to detect the luciferase expression . In the meantime, five classes of organic compounds that may affect the balance of thyroid hormones in the body were determined. Results As the concentration of the test solution increased, the luciferase expression decreased and the activity of T3 decreased. Linear expression of Luciferase at the concentrations of 0.01, 10.00 and 20.00 mg dust / ml was lower than 5 nmol / L T3 positive control group, P respectively 0.012,0.002. Chemical analysis showed that the contents of common endocrine disrupting chemicals in indoor dusts were: polycyclic aromatic hydrocarbons 0.2 ~ 3.7μg / g, phthalates undetectable (~ 117.4μg / g), polybrominated diphenyl ethers ND ~ 1.0μg / g, PCBs 1.0 ~ 25.0 ng / g, phenols ND ~ 32.5μg / g. Conclusion The indoor dust organic extract can interfere with the thyroid hormone activity, suggesting that the early indoor dust exposure in children has a potential impact on the neurodevelopment of children.