【作者】 潘红梅；

【导师】 吴德生；

【作者基本信息】 四川大学 ， 劳动卫生与环境卫生学， 2004， 硕士

【摘要】 为研究FRTL-5细胞应用于环境甲状腺素干扰物体外甄别方法的可行性,首先进行细胞毒性实验,了解6种环境化学物amitrol, PCP, ETU, pendimethalin, prodiamine, nitrofen对FRTL-5细胞的毒性,然后分别进行放免法测培养液中甲状腺球蛋白的含量;免疫细胞化学法测细胞胞浆内甲状腺球蛋白及细胞核内TTF-1水平变化;酶细胞化学法测细胞胞浆内甲状腺过氧化物酶活性变化。 在细胞毒性实验中,amitrol各剂量均无显著细胞毒性;当PCP浓度超过0.0027ng/ul,ETU为360ng/ul,pendimethalin为2500ng/ul,prodiamine为0.04ng/ul,nitrofen为100ng/ul时,有显著细胞毒性。 在放免实验中测甲状腺球蛋白时,amitrol, ETU, PCP在中、高剂量显著降低培养液中的TG浓度,pendimethalin, prodiamine在中、高剂量显著增加培养液液中TG浓度,nitrofen各剂量组对培养液中TG浓度无显著影响。 在免疫细胞化学实验中测胞浆内TG时,amitrol在高剂量显著降低胞浆内TG水平,pendimethalin在高剂量显著增强胞浆内TG水平,其它几种环境化学物均对胞浆内TG水平无显著影响。测细胞核内TTF-1时,PCP, pendimethalin在中、高剂量显著降低核内TTF-1表达水平,ETU各剂量组均显著降低核内TTF-1水平,其它几种环境化学物对核更多还原

【Abstract】 To study the feasibity of using FRTL-5 cell in screening methods in vitro of environmental thyroid hormone disruptors. First, study the cytotoxicity of amitrol, pendimethalin, nitrofen,PCP,ETU,Prodiamine,using MTT. Then using RIA(radioimmunoassay) to test the concentration of TG in culture; using ICC to test the level of TG in FTRL-5 cell and the level of TTF-1 in nuclear; using enzymecellchemistry to test the activity of thyroid peroxidase in FRTL-5 cell.In MTT test, there is distinct cytotoxicity in all dose of amitrol. When the concentration of PCP exceeds 0.0027ng/ul;ETU exceeds 360ng/ul;pendiametnalin exceeds 2500ng/ul;prodiamine exceeds 0.04ng/ul;nitrofen exceeds 100ng/ul,trere is distinct cytotoxicity.In RIA, the concentration of TG in culture is decreased distinctly in medium and high dose of amtrol, ETU, PCP. The concentration of TG in culture is increased distinctly in medium and high dose of pendimethalin, prodiamine. All dose of nitrofen have not distinct effects on the concentration of TG in culture.In ICC test, the concentration of TG in cell is descreased distinctly in high dose of amitrol and pendimethalin. The other environmental chemicals have not obvious effects on the concentration of TG in cell. The level ofTTF-1 in nuclear is obviously decreased in high dose of PCP, pendimethalin and all dose of ETU. The other environmental chemicals have not obvious effects on the level of TTF-1 in nuclear.In enzymecellchemistry test, the activity of TPO is decreased obviously in high dose of amitrol and pendimethalin. The other environmental chemicals have not obvious effects on the activity of TPO in cell.The study obtained expected results and further evidenced the feasibity of using FRTL-5 cell in screening methods in vitro of environmental thyroid hormone disruptors. The obtained results will be the base of future research.更多还原