过量碘诱导甲状腺细胞焦亡与桥本氏甲状腺炎相关机制研究  

Excessive iodine-induced pyroptosis of thyroid cells and related mechanisms of Hashimoto's thyroiditis

作　　者：王森塬 谭丽艳[1] 于学静[1] Senyuan Wang;Liyan Tan;Xuejing Yu(Department of Endocrinology,the First Affiliated Hospital of Jiamusi University,Jiamusi 154000,China)

机构地区：[1]佳木斯大学附属第一医院内分泌科,黑龙江佳木斯154000

出　　处：《国际免疫学杂志》2021年第6期609-614,共6页International Journal of Immunology

摘　　要：目的探究过量碘对甲状腺细胞焦亡的影响及焦亡与桥本氏甲状腺炎的相关性。方法将2019年1月至2020年2月佳木斯大学附属第一医院普外科手术取下的甲状腺腺瘤伴桥本甲状腺炎组织(实验组)及甲状腺结节周围正常组织样本(对照组)各6例立即保存于-80℃,提取RNA后实时荧光定量逆转录聚合酶链反应(quantificational real-time polymerase chain reaction,qRT-PCR)检测细胞焦亡相关因子NLR家族Pyrin域蛋白3(NLR family pyrin domain containing protein 3,NLRP3)、半胱氨酸天冬氨酸蛋白酶-1(cysteine-aspartic acid protease-1,caspase-1)以及白细胞介素-1β(interleukin-1β,IL-1β)的表达。应用不同浓度的碘化钾(1、10、20、50以及100 mmol/L)诱导甲状腺上皮细胞,12 h后进行CCK-8检测筛选最佳实验浓度并观察甲状腺细胞形态的变化以及NLRP3重组蛋白、caspase-1以及IL-1β的表达变化。加入caspase-1抑制剂后进一步观察NLRP3、caspase-1以及IL-1β的表达变化。结果桥本甲状腺炎患者手术组织标本中NLRP3、caspase-1以及IL-1β的表达水平升高[NLRP3:(3.58±0.98)比(1.02±0.29);caspase-1:(3.37±0.46)比(1.03±0.47);IL-1β:(3.48±0.87)比(1.09±0.61),t值分别为9.863、9.057、7.875,P值均<0.05];在细胞实验中,以50 mmol/L碘化钾处理甲状腺上皮细胞时的细胞形态变化最显著,细胞中的NLRP3,caspase-1以及IL-1β的mRNA[NLRP3:(2.87±0.82)比(1.04±0.39);caspase-1:(2.85±0.78)比(1.06±0.47);IL-1β:(3.10±1.01)比(1.06±0.24),t值分别为7.702、5.121、6.263,P值均<0.05]及蛋白质[NLRP3:(3.21±1.13)比(1.04±0.61);caspase-1:(2.78±0.29)比(1.06±0.94);IL-1β:(2.99±0.56)比(1.24±0.27),t值分别为8.194、10.290、14.090,P值均<0.05]表达水平升高,加入caspase-1抑制剂后这种变化被一定程度的逆转。结论过量碘通过影响甲状腺细胞的焦亡参与桥本氏甲状腺炎的发生发展。Objective To explore the effect of excessive iodine on pyroptosis of thyroid cells and the correlation with Hashimoto's thyroiditis.Methods The tissue samples of 6 cases of thyroid adenoma with hashimoto's thyroiditis(experimental group)and normal tissue samples around thyroid nodule(control group)were taken from the General Surgery Department of the First Affiliated Hospital of Jiamusi University between January 2019 and February 2020,and the tissue samples were immediately stored at-80℃.The RNA was extracted and the expression of pyroptosis-related factors NLR family pyrin domain containing protein 3(NLRP3),cysteine-aspartic acid protease-1(caspase-1)and interleukin-1β(IL-1β)were detected.Different concentrations of potassium iodide(1,10,20,50 and 100 mmol/L)were used to induce thyroid epithelial cells.12 h later,CCK-8 was used to screen the optimal concentration and observe the changes of thyroid cell morphology,the expression of NLRP3 recombinant protein,caspase-1 and IL-1βwere deteted at the same time.The expression changes of NLRP3,Caspase-1 and IL-1βwere further observed after the addition of caspase-1 inhibitors.Results The expressions of NLRP3,caspase-1,and IL-1βincreased in Hashimoto's thyroiditis patients were signficantly increased[NLRP3:(3.58±0.98)vs(1.02±0.29);Caspase-1:(3.37±0.46)vs(1.03±0.47);IL-1β:(3.48±0.87)vs(1.09±0.61),t values were 9.863,9.057,7.875,respectively,all P values<0.05].In cell experiments,the morphological changes of thyroid epithelial cells were most significant when treated with 50 mmol/L potassium iodide.The mRNA and protein levels of NLRP3,caspase-1 and IL-1βin thyroid epithelial cells were both increased[mRNA:NLRP3:(2.87±0.82)vs(1.04±0.39);Caspase 1:(2.85±0.78)vs(1.06±0.47);IL-1β:(3.10±1.01)vs(1.06±0.24),t values were 7.702,5.121,6.263,respectively,all P values<0.05,protein:NLRP3:(3.21±1.13)vs(1.04±0.61);caspase-1:(2.78±0.29)vs(1.06±0.94);IL-1β:(2.99±0.56)vs(1.24±0.27),t values were 8.194,10.290,14.090,respectively,all P values<0.05].These changes

关 键 词：过量碘 细胞焦亡 桥本氏甲状腺炎 

分 类 号：R581.4[医药卫生—内分泌] R736.1[医药卫生—内科学]