环境内分泌干扰物双酚A对小鼠甲状腺滤泡上皮细胞增殖和凋亡的影响  

Effects of environmental endocrine disruptor bisphenol A on proliferation and apoptosis of mouse thyroid follicular epithelium cells

作　　者：刘泽兵[1] 王丽[1] 叶宣光[1] 于秀杰[2] 张友元[1] 

机构地区：[1]复旦大学附属金山医院病理科,上海201508 [2]天津医科大学内分泌研究所卫生部激素与发育重点实验室,天津300070

出　　处：《中国病理生理杂志》2013年第6期1076-1080,共5页Chinese Journal of Pathophysiology

基　　金：上海市金山科学技术创新项目(No.2011-3-12);上海市卫生局面上科研基金资助项目(No.20100051)

摘　　要：目的:探讨环境内分泌干扰物双酚A(BPA)对小鼠甲状腺原代培养细胞增殖和凋亡的影响。方法:取雌性BALB/c nu/nu小鼠甲状腺组织,采用胶原酶I/中性蛋白酶联合消化并进行滤泡上皮细胞培养,Westernblotting检测甲状腺球蛋白表达对细胞进行鉴定。采用不同浓度的BPA干预稳定培养48 h的小鼠甲状腺原代培养的滤泡上皮细胞,作用24 h后,光学显微镜观测干预前后细胞生长状态的变化,流式细胞术分析细胞增殖和凋亡,real-time PCR法和免疫细胞化学染色法测定干预前后肿瘤坏死因子相关凋亡诱导配体受体1(TRAIL-R1)mRNA和蛋白的表达变化。结果:原代培养的小鼠甲状腺滤泡上皮细胞随BPA刺激浓度的增加,生长状态呈现先促进后抑制的趋势;0.01～0.1μmol/L BPA作用24 h后,细胞增殖呈剂量依赖性增长,0.1μmol/L BPA可显著刺激细胞增殖(P<0.05),而1μmol/L BPA刺激后则表现为细胞增殖水平显著下降(P<0.05);0.1μmol/L BPA刺激后凋亡指数减低(P<0.05);0.1μmol/L和1μmol/L BPA刺激后TRAIL-R1 mRNA表达均显著下调(均P<0.05),蛋白表达与mRNA表达结果趋势一致。结论:低、中浓度BPA可刺激原代培养的小鼠甲状腺滤泡上皮细胞增殖并抑制细胞凋亡,而高浓度BPA对细胞主要表现为毒性损害作用。BPA对细胞凋亡的影响可能与TRAIL-R1相关途径有关。AIM： To investigate the effect of environmental endocrine disruptor bisphenol A （BPA） on the proliferation and apoptosis of primarily cultured mouse thyroid follicular epithelium cells （TFECs）. METHODS： The thy- roids of female BALB/c nu/nu mice were digested by the combination of collagenase I and dispase. The isolated TFECs were cultured and identified by Western blotting. Mter cultured for 48 h, the TFECs were stimulated with different concen- trations of BPA for 24 h. The morphological changes of TFECs were observed under light microscope. The cells were diges- ted and harveste, and the proliferation and apoptosis were determined by the application of flow cytometry. The expression of TNF-related apoptosis-inducing ligand receptor 1 （ TRAIL-R1 ） at protein and mRNA levels was investigated by the methods of immunocytochemistry and real-time PCR, respectively. RESULTS ： With the increase in BPA concentration, the growth of TFECs was firstly promoted and then inhibited. Dose-dependent proliferation was exhibited by the stimulation of BPA at 0.01-0.1 μmol/L, and proliferation index （PI） was significantly enhanced by 0. 1 μmol/L BPA and attenuated by 1 μmol/L BPA. The apoptotic index （AI） was significantly decreased by 0.1 μmol/L BPA. The expression of TRAIL-R1 at mRNA and protein levels was drastically down-regulated by 0.1 μ mol/L and 1 μ mol/L BPA. CONCLUSION： Prolifera- tion of TFECs is promoted by the stimulation of low to medium concentrations of BPA, while high concentration of BPA mainly exhibits a toxic effect. Additionally, the effect of BPA on apoptosis is potentially mediated through TRAIL-Rl-relat- ed pathway.

关 键 词：双酚A 细胞增殖 细胞凋亡 甲状腺 

分 类 号：R736.1[医药卫生—肿瘤]