基于NLRP3/Caspase-1/GSDMD通路探讨自身免疫性甲状腺炎伴抑郁症模型的制备与评价  

Establishment and evaluation of mouse models of autoimmune thyroiditis with depression based on the NLRP3/Caspase-1/GSDMD pathway

作　　者：李铮铮 刘子玉 王智民[3] 金哲[2] 赵卓 杨潇[2] LI Zhengzheng;LIU Ziyu;WANG Zhimin;JIN Zhe;ZHAO Zhuo;YANG Xiao(Liaoning University of Traditional Chinese Medicine Graduate School,Shenyang 110847,China;the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学研究生学院,沈阳110847 [2]辽宁中医药大学附属第二医院,沈阳110034 [3]辽宁中医药大学附属医院,沈阳110032

出　　处：《中国实验动物学报》2024年第8期955-964,共10页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金面上项目(82274455),国家自然科学基金青年基金项目(82104805);辽宁省教育厅重点攻关项目(JYTZD2023196)。

摘　　要：目的探讨自身免疫性甲状腺炎伴发抑郁症动物模型的制备与评价,并基于NOD样受体蛋白-3(NOD-like receptor protein 3,NLRP3)/含半胱氨酸的天冬氨酸蛋白水解酶-1(cysteinyl aspartate specific proteinase-1,Caspase-1)/消皮素D(gasdermin D,GSDMD)通路加以验证。方法32只NOD.H-2H4小鼠随机分为正常组(N组)、抑郁组(DP组)、自身免疫性甲状腺炎伴抑郁症组(AIT+DP组)、自身免疫性甲状腺炎组(AIT组),每组8只。N组正常饲养,DP组采取5周慢性不可预知温和刺激(chronic unpredictable mild stress,CUMS),AIT组予0.05%碘化钠水溶液建立自身免疫性甲状腺炎模型,AIT+DP组在建立AIT动物模型基础上施加5周CUMS建立AIT+DP动物模型。通过观测小鼠甲状腺组织结构及淋巴细胞浸润情况和血清甲状腺过氧化物酶抗体(thyroid peroxidase antibody,TPOAb)和甲状腺球蛋白抗体(anti-thyroid autoantibodies,TGAb)水平评价小鼠自身免疫性甲状腺炎模型是否制备成功;通过测定体重、糖水偏好率、旷场行为学(中央象限时间、中央象限比例、站立次数、排便次数、毛发梳理时间),大脑皮质、海马病理变化及大脑皮质小胶质细胞焦亡相关蛋白水平评价小鼠抑郁状态。模型小鼠同时符合上述自身免疫性甲状腺炎与抑郁症相关指标检测,则表明AIT+DP动物模型制备成功。结果与N组比较,AIT组与AIT+DP组血清TGAb、TPOAb水平显著增加(P<0.01),甲状腺可见大量炎细胞浸润,DP组与AIT+DP组小鼠中央象限时间、中央象限比例、站立次数、排便次数、毛发梳理时间有不同程度降低,大脑皮质神经胶质细胞增多,神经元细胞减少,伴有部分细胞核萎缩,NLRP3、IL-1β、Caspase-1、GSDMD-N蛋白表达水平显著上调,AIT+DP组尤为明显(P<0.01)。结论0.05%碘化钠水溶液与CUMS可较好地模拟AIT+DP模型动物外在表现与内在指标变化,可为AIT+DP疾病的研究提供动物模型参考。Objective To investigate the preparation and evaluation of animal models of depression associated with autoimmune thyroiditis,and to verify the NLRP3/Caspase-1/GSDMD pathway based on this condition.Methods 32 NOD.H-2H4 mice were randomly divided into a normal(N)group,depression(DP)group,autoimmune thyroiditis with depression(AIT+DP)group,autoimmune thyroiditis(AIT)group,with 8 animals in each group.The N group was fed normally,the DP group was subjected to chronic unpredictable mild stress(CUMS)for 5 weeks,the AIT group was given 0.05%sodium iodide water to establish an autoimmune thyroiditis model,and the AIT+DP group was subjected to 5 weeks of CUMS to establish the AIT animal model.We evaluated whether the mouse autoimmune thyroiditis model had been successfully prepared by observing the thyroid tissue structure,lymphocyte infiltration,and serum TGAb and TPOAb levels.Changes in body weight,sugar water preference rate,open field behavior(central quadrant time,central quadrant time proportion,standing rate,frequency of defecation,and hair grooming time),and hippocampal pathological changes were used to evaluate the depression status of the mice.When the model mice met the above-mentioned indicators related to depression and autoimmune thyroiditis,the AIT+DP animal model was considered successfully prepared.Results Compared with the levels in the N group,the AIT group’s and AIT+DP group’s serum TGAb and TPOAb levels were significantly increased(P<0.01),and a large number of inflammatory cells had infiltrated the thyroid gland.The central quadrant time and central quadrant time proportion,standing rate,frequency of defecation,and hair grooming time were reduced to varying degrees in the DP group and AIT+DP group.In addition,the numbers of glial cells in the cerebral cortex increased and neuronal cells decreased,accompanied by some nuclear atrophy,and the expression levels of NLRP3,IL-1β,Caspase-1,and GSDMD-N significantly increased,especially in the AIT+DP group(P<0.01).Conclusions 0.05%sodium iodide water an

关 键 词：自身免疫性甲状腺炎 抑郁症 模型评价 NLRP3/Caspase-1/GSDMD通路 细胞焦亡 

分 类 号：Q95-33[生物学—动物学]