摘要
[目的 ]研究磺胺二甲嘧啶对FRTL 5细胞株合成、分泌甲状腺球蛋白的影响 ,并探讨FRTL 5细胞株用于筛选环境甲状腺素干扰物的可能性。 [方法 ]FRTL 5细胞分别经 0 .5、1.0、2 .0、4.0、和 8.0 μg/ml磺胺二甲嘧啶染毒处理48h后 ,用MTT实验检测磺胺二甲嘧啶对细胞活力的影响以确定染毒浓度 ;以MTT实验最大无作用浓度作为最高染毒浓度 ,另设中、低浓度共 3组染毒细胞 ,用放射免疫法测定染毒 48h后培养液中细胞分泌甲状腺球蛋白的浓度 ;用活细胞爬片的免疫组化染色观察最高浓度处理组细胞胞质中合成甲状腺球蛋白的表达水平 ,并用透射电镜观察细胞的超微结构。[结果 ]根据MTT实验 ,确定染毒浓度为 0 .5、1.0和 2 .0 μg/ml,随磺胺二甲嘧啶浓度的增大 ,FRTL 5细胞培养液中甲状腺球蛋白的浓度逐渐降低 ,其中 1.0 μg/ml浓度组与对照组相比差异有统计学意义 (P <0 .0 5 ) ,2 .0 μg/ml浓度组培养液中未能检出甲状腺球蛋白 ;免疫组化染色可见染毒组细胞胞质中合成的甲状腺球蛋白表达显著减弱 ,图像分析结果显示其灰度值的差异有统计学意义 (P <0 .0 5 ) ;细胞超微结构观察发现染毒组细胞出现粗面内质网扩张。 [结论 ]通过损伤甲状腺滤泡细胞的粗面内质网 ,而影响甲状腺球蛋白的合成与分泌 。
Objective] To study the effect of sulfamethazine on producing and secreting thyroglobulin by FRTL-5 cells, and to explore the probability of FRTL-5 cell to be used in screening environmental thyroid hormone disruptors in vitro. [Methods] The cells were treated with 0.5,1.0,2.0,4.0,and 8.0 μg/ml sulfamethazine for 48 hours, and then cell viability was determined using the MTT dye assay. The treatment concentrations were designed according to the results of MTT experiment. The thyroglobulin concentration in medium was detected by radioimmunoassay 48 hours after treatment;the expression of thyroglobulin in FRTL-5 cells was assessed by immunohistochemical technique;ultrastructure of the cells was also observed. [Results] According to MTT experiment,cells were treated with 0.5,1.0,and 2.0 μg/ml sulfamethazine. Sulfamethazine decreased the thyroglobulin levels in medium when compared with the control,especially at the concentration of 1.0 μg/ml (P<0.05); thyroglobulin was not detectable at the concenation of 2.0 μg/ml. Immunohistochemistry determination showed that the expression of thyroglobulin inside the cells was weakened in the treated cells,and there was significant difference between the gray value of control and treated cells. The treated cells also had expanded rough endoplasmic reticulum. [Conclusion] One of the main mechanism of sulfamethazine disrupting the bio-function of thyroid is to damage the rough endoplasmic reticulum of thyroid follicular cells and introducing FRTL-5 cells into screening thyroid hormone disruptors in vitro is feasible.
出处
《环境与职业医学》
CAS
北大核心
2004年第3期182-185,共4页
Journal of Environmental and Occupational Medicine
基金
国家自然科学基金重点课题 (编号 :30 0 30 1 2 0 )
纽约中华医学基金资助
