MicroRNA-155在碘致NOD.H-2h4小鼠自身免疫性甲状腺炎发病中的变化研究  

Study of microRNA-155 changes in iodine-induced autoimmune thyroiditis model of NOD. H-2 h4 Mice

作　　者：刘通[1] 于振乾[1] 邹红锦 刘姗姗[1] 范晨玲[1] 滕卫平[1] 单忠艳[1] 李玉姝(指导)[1] 

机构地区：[1]中国医科大学附属第一医院内分泌科,内分泌研究所,辽宁省内分泌疾病重点实验室,沈阳110001

出　　处：《中国免疫学杂志》2015年第7期936-941,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(No.81471003)

摘　　要：目的:观察高碘诱发NOD.H-2 h4小鼠发生自发性自身免疫性甲状腺炎(SAT)动物模型中Th1及Th2细胞的动态变化及甲状腺、脾脏、肝脏、肾脏中mi R-155的动态改变,探讨mi R-155对碘致自身免疫性甲状腺炎(AIT)的可能影响及作用机制。方法:4周龄NOD.H-2 h4雌性小鼠随机分为对照组及高碘饮水组。高碘组饲以含0.05%碘化钠的无菌水,对照组饲以无菌双蒸水,于实验开始后第0、2、4、6、8、10、12周麻醉处死。留取甲状腺,制备石蜡切片行HE染色;酶联免疫吸附试验测定血清甲状腺球蛋白抗体(Tg Ab)水平;流式细胞仪检测脾脏单个核细胞悬液Th1、Th2细胞表达水平;实时定量RT-PCR检测脾细胞中Th1和Th2细胞转录因子T-bet及GATA-3,细胞因子IFN-γ及IL-4的m RNA表达水平及甲状腺、脾脏、肝脏、肾脏mi RNA-155表达水平。结果:发生SAT的小鼠血清Tg Ab滴度较对照组升高(P<0.05)。脾细胞中Th1细胞比例及T-bet,IFN-γm RNA表达量自6周后较对照组升高(P<0.05)。脾脏、甲状腺miR-155表达量分别自8周及10周后较对照组升高(P<0.05)。脾脏mi R-155水平与Tg Ab滴度显著正相关(r=0.825,P<0.01),与Th1细胞比例显著正相关(r=0.872,P<0.01);甲状腺mi R-155水平与TgAb滴度显著正相关(r=0.575,P<0.01);与Th1细胞比例显著正相关(r=0.642,P<0.01)。结论:发生SAT的NOD.H-2 h4小鼠脾脏Th1细胞比例显著升高,提示Th1细胞可能参与该模型AIT的发生发展,占免疫优势地位。脾脏及甲状腺mi R-155水平与Tg Ab滴度及Th1细胞比例显著正相关,提示mi R-155可能通过影响Th1细胞在AIT的发生发展中发挥作用。Objective：To investigate the dynamic changes of Th1 cell and Th2 cell,the expression of micro RNA-155 in thyroid gland,spleen,liver and kidney in NOD. H-2 h4 mice with iodine-induced autoimmune thyroiditis,and explore the potential effect and mechanism of micro RNA-155 on AIT induced by iodine. Methods： NOD. H-2 h4 mice at 4 weeks of age were randomly divided into control and iodine-treated（ HI） groups,mice in HI group were fed with 0. 05% Na I,and the control group received sterile water. Animals were sacrificed at the time point of 0,2,4,6,8,10 and 12 weeks. Thyroid tissues were removed and the severity of lymphocytic infiltration in thyroids was observed using HE-stained thyroid sections. The serum Tg Ab titers were measured by ELISA. The percentage of CD4＋IFN-γ＋T cells（ Th1） and CD4＋IL-4＋T cells（ Th2） in splenic mononuclear cells were detected by multicolor flow cytometry. The expression of Th1 and Th2 cell related transcription factor T-bet and GATA-3 m RNA,the related cytokines IFN-γ and IL-4 m RNA,and micro RNA-155 in thyroid gland,spleen,liver and kidney were measured by RT-PCR. Results： Serum Tg Ab titers of SAT mice increased obviously than the control group（ P 〈0. 05）. Higher number of CD4＋IFN-γ＋T cells in the splenic mononuclear cells,and higher expressions of T-bet and IFN-γ m RNA was observed in SAT mice after driking Na I solution 6 weeks（ P 〈0. 05）. Compared with the control group,mice with AIT had rised expression of micro RNA-155 in spleen and thyroid after 8 weeks and 10 weeks,respectivley（ P 〈0. 05）. In spleen and thyroid tissue of SAT mice,there were positive relationships between the expression of mi R-155 and the titers of serum Tg Ab（ r = 0. 825,P 〈0. 01 and r = 0. 872,P 〈0. 01）,which were the same with the percentage of Th1 cell（ r = 0. 575,P 〈0. 01 and r = 0. 642,P 〈0. 01）. Conclusion： The data indicate that the increase of CD4＋IFN-γ＋T cells plays an important role in the development of AIT. The expression of micr

关 键 词：NOD.H-2 h4小鼠 自身免疫性甲状腺炎 microRNA-155 TH1细胞 

分 类 号：R581.4[医药卫生—内分泌]