Thyroid hormone disrupting activity of house dust and related pollutants
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摘要: 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pUAS-tk-Luc、表达质粒Gal4-L-TRβ和SofastTM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0.2~3.7 μg/g,邻苯二甲酸酯类未检出(not detected,ND)~117.4 μg/g,多溴联苯醚类ND~1.0 μg/g,多氯联苯类1.0~25.0 ng/g,酚类ND~32.5 μg/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。Abstract: Objective To detect the thyroid hormone disrupting activity of house dust extract with the reporter transcriptional activation assay. Methods The restructured reporter plasmid pUAS-tk-Luc, expression vector Gal4-L-TRβand transfection reagent SofastTM were transfected into HEK-293 cells. Different concentrations of house dust extract, and five nmol/L triiodothyronine (T3) were added into the culture medium. The expression of the luciferase was detected by the luciferase reporter gene detector. Simultaneously, 5 families of organic compounds with possibility to disrupting the thyroid hormone were analyzed. Results The one-way ANOVA linear trend test showed that the luciferase expression decreased with the increased concentrations of the extract, which indicated the decreased activity of the T3 with P<0.001. The luciferase expression of the 10.00 and 20.00 mg dust/mL groups was significantly lower than that of the 5 nmol/L T3 group (P=0.012 and 0.002, respectively). Chemical analysis showed that polycyclic aromatic hydrocarbons, phalate esters, polybrominated diphenyl ethers, polychlorinated biphenyl and phenols were 0.2-3.7 μg/g, not detected-117.4 μg/g, not detected-1.0 μg/g, 1.0-25.0 ng/g and not detected-32.5 μg/g, respectively. Conclusions The activity of the thyroid hormone could be inhibited by house dust extract by disrupting the combination with the thyroid hormone receptor, which indicated that the house dust exposure in early childhood might cause potential neuro developmental influence.
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Key words:
- Thyroid hormones /
- Endocrine disruptors /
- Epidemiologic studies
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