过氯酸铵及碘化钾联合作用对人甲状腺细胞凋亡及氧化应激的影响

Combined effects of exposure to ammonium perchlorate and potassium iodide on apoptosis and oxidative stress in the human thyroid cells

目的研究过氯酸铵(AP)及碘化钾(KI)联合作用对人甲状腺细胞(nthy-ori3-1)氧化应激及诱导凋亡的影响。方法体外培养nthy-ori3-1细胞,不同浓度AP(0、5、10、20、40mmol/L)及KI(5mmol/L)联合染毒后培养24h,CCK8法检测nthy-ori3-1细胞的增殖活力,流式细胞仪检测细胞凋亡率及细胞内活性氧(ROS)产生情况,同时测定氧化损伤指标丙二醛(MDA)、过氧化氢酶(CAT)活力。结果AP抑制nthy-ori3-1细胞增殖具有明显的剂量依赖性。与对照组比较,各组细胞内ROS生成量随着染毒剂量的增加出现降低趋势,除5mmol/L AP剂量组外,差异均有统计学意义(P<0.05),且AP与KI联合染毒剂量组细胞内ROS生成量比单纯高剂量AP染毒(40mmol/L)细胞内ROS生成量有所降低,差异有统计学意义(P<0.05);较高剂量AP染毒(40mmol/L)甲状腺细胞,MDA生成量比对照组有明显升高,差异有统计学意义(P<0.01);各染毒剂量组细胞内CAT活力比对照组有升高的趋势,但差异无统计学意义(P>0.05)。结论 AP和KI联合作用可致nthy-ori3-1细胞出现氧化应激,且二者之间存在明显的剂量-反应关系,碘在一定程度上可缓解AP对甲状腺细胞的氧化应激反应。

Objective To explore the combined effects of ammonium perchlorate（AP）and potassium iodide（KI）on the oxidative stress and apoptosis in Nthy-ori3-1 thyroid cells. Methods The Nthy-ori3-1 cells cultured at 37 ＂C, 5% CO2in vitro, were exposed to different concentrations of AP （10,20,40 mmol/L） and KI（5,25 mmol/L）for 24 h, then the cell proliferation activity was detected by the CCK8 assay, while the apoptosis and intracellular generated reactive oxygen species （ROS）were measured by flow cytometry, simultaneously with the oxidative damage indicators malondialdehyde（MDA）and catalase（CAT）activity being detected. Results The significant dose-dependence relationship was found in the inhibition of Nthy-ori3-1 cell proliferation induced by AP. Compared with the control group, the ROS generation of the cells in each group appeared to decrease with dose increase except for the 5mmol/LAP dose group, and the differences were statistically significant（P〈0. 05）; The ROS generation of the thyroid cells exposed to combination toxic effect of AP and KI decreased, when compared with that of simple high dose AP exposure （40mmol/L）, and the difference was statistically significant （P 〈 0.05）; Compared with the control group, there was significantly increase in the MDA generation of thyroid cells exposed to higher dose of AP （40mmol/L）, and the difference was statistically significant （P〈0. 001） ; The CAT activity of the thyroid cells exposed to any doses was elevated with the comparison of the control group, hut the difference was not statistically significant （P〉 0. 05）. Conclusions AP might lead to oxidative stress on the Nthy-ori3-1 cell, as well as the significant dose-response relationship between the dose and effect. Iodine might alleviate the oxidative stress to some extent, corresponding to thyroid cells induced by AP.