过量碘通过p38和细胞间黏附分子-1促进小鼠实验性自身免疫性甲状腺炎的发生发展  

Excessive iodine promotes the occurrence and development of experimental autoimmune thyroiditis in mice through p38 and intercellular adhesion molecule-1

作　　者：李亚楠[1] 沈洪婷[1] 王明君 陈勋[1] 马静[1] 甘培春[1] 张强[1] 张金梅 曹晓晓 孟献亚[1] Li Yanan;Shen Hongting;Wang Mingjun;Chen Xun;Ma Jing;Gan Peichun;Zhang Qiang;Zhang Jinmei;Cao Xiaoxiao;Meng Xianya(Department of Endemic Disease,Qinghai Institute for Endemic Disease Prevention and Control,Xining 811602,China;National Health Commission Key Laboratory of Trace Elements,National Institute of Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]青海省地方病预防控制所地方病科,西宁811602 [2]中国疾病预防控制中心营养与健康所国家卫生健康委微量元素重点实验室,北京100050

出　　处：《中华地方病学杂志》2022年第4期270-276,共7页Chinese Journal of Endemiology

摘　　要：目的探讨过量碘致小鼠实验性自身免疫性甲状腺炎(EAT)发生的分子机制。方法选取60只雌性非肥胖糖尿病(NOD)小鼠,按照体质量[(25±3)g]采用随机数字表法分为5组,每组12只:对照组(A组),10倍高碘组(B组),100倍高碘组(C组),1000倍高碘组(D组),1000倍高碘联合聚肌胞苷酸[Poly(I:C)]组(E组)。实验周期为16周,各组小鼠分别饮用碘化钠(NaI)含量为0.000、0.005、0.050、0.500、0.500 mg/L的纯净水;E组小鼠分别于实验第7周和第15周腹腔注射Poly(I:C)。实验第16周末解剖小鼠,取血样和甲状腺组织。采用酶联免疫吸附测定(ELISA)法检测血清甲状腺功能指标[促甲状腺激素(TSH)、游离三碘甲状腺原氨酸(FT_(3))、游离甲状腺素(FT_(4))、甲状腺过氧化物酶抗体(TPOAb)]水平;采用苏木素-伊红(HE)染色观察甲状腺组织病理变化;采用转录组测序(RNA-seq)技术进行甲状腺组织差异表达基因分析,对差异表达基因进一步进行KEGG通路分析;采用实时荧光定量PCR(qPCR)、免疫印迹法(Western blot)分别检测甲状腺组织p38、细胞间黏附分子-1(ICAM-1)、趋化因子10(CXCL10)mRNA和蛋白水平。结果5组小鼠血清TSH(ng/ml:6.53±0.86、6.61±0.82、7.68±0.55、7.93±0.60、8.73±1.60),FT_(3)(pg/ml:59.35±10.16、53.73±10.96、46.19±8.03、41.01±8.67、34.21±11.75),FT_(4)(pg/ml:136.74±10.06、124.33±14.34、101.80±6.78、91.37±6.75、73.29±17.31),TPOAb水平(U/ml:130.81±24.53、145.47±28.89、166.52±41.59、199.78±42.19、201.99±44.03)比较,差异均有统计学意义(F=4.77、4.96、23.12、3.68,P均<0.05)。与A组比较,C、D、E组小鼠血清TSH水平均较高,B、C、D、E组FT_(3)、FT_(4)水平均较低,D、E组TPOAb水平均较高,差异均有统计学意义(P均<0.05)。HE染色显示,D、E组甲状腺滤泡病变程度较严重,均出现了EAT的表型。将差异表达基因进行KEGG通路分析,B、C、D、E组与A组比较,发现8条与甲状腺自身免疫反应和炎症反应相关的代谢�Objective To investigate the molecular mechanism of excessive iodine induced experimental autoimmune thyroiditis(EAT)in mice.Methods Sixty female non-obese diabetic(NOD)mice were selected and divided into 5 groups according to body weight[(25±3)g]via the random number table method,with 12 mice in each group:control group(group A),10-fold high iodine group(group B),100-fold high iodine group(group C),1000-fold high iodine group(group D)and 1000-fold high iodine combined with polyinosinic acid-polycytidylic acid[Poly(I:C)]group(group E).The experiment period was 16 weeks.Mice in each group drank purified water with sodium iodine(NaI)content of 0.000,0.005,0.050,0.500 and 0.500 mg/L,respectively;mice in group E were intraperitoneally injected with Poly(I:C)at week 7 and week 15,respectively.At the end of the 16th week,mice were dissected and blood samples and thyroid tissue were taken.The levels of serum thyroid function indexes[thyroid stimulating hormone(TSH),free triiodothyronine(FT_(3)),free thyroxine(FT_(4)),and thyroid peroxidase antibody(TPOAb)]were detected by enzyme-linked immunosorbent assay(ELISA);the pathological changes of thyroid tissue were observed after hematoxylin-eosin(HE)staining;differentially expressed genes in thyroid tissue were detected by RNA-sequencing(RNA-seq),and analyzed by KEGG pathway;mRNA and protein levels of p38,intercellular adhesion molecule-1(ICAM-1)and chemokine 10(CXCL10)in thyroid tissue were detected by real-time fluorescence quantitative PCR(qPCR)and Western blotting,respectively.Results There were statistically significant differences in serum levels of TSH(ng/ml:6.53±0.86,6.61±0.82,7.68±0.55,7.93±0.60,8.73±1.60),FT_(3)(pg/ml:59.35±10.16,53.73±10.96,46.19±8.03,41.01±8.67,34.21±11.75),FT_(4)(pg/ml:136.74±10.06,124.33±14.34,101.80±6.78,91.37±6.75,73.29±17.31),and TPOAb(U/ml:130.81±24.53,145.47±28.89,166.52±41.59,199.78±42.19,201.99±44.03)among the 5 groups of mice(F=4.77,4.96,23.12,3.68,P<0.05).Compared with group A,the serum TSH levels of mice in groups

关 键 词：碘 小鼠 实验性自身免疫性甲状腺炎 转录组测序 P38 细胞间黏附分子-1 

分 类 号：R581.4[医药卫生—内分泌]