Wnt7a/β-连环蛋白/自身免疫调节因子信号通路参与1型糖尿病的发生  

Wnt7a/β-catenin/autoimmune regulator signaling pathway involved in the occurrence of type 1 diabetes

作　　者：李远迪 马静怡 余佳珊 何可可 文廷浩 高杰 李红 苏敏 胡蓉[1,3] LI Yuan-di;MA Jing-yi;YU Jia-shan;HE Ke-ke;WEN Ting-hao;GAO Jie;LI Hong;SU Min;HU Rong(Department of Histology and Embryology,Basic Medical College,Guizhou Medical University,Guiyang 550025,China;National Joint Local Engineering Laboratory for Cell Engineering and Biomedicine Technique,Guizhou Province Key Laboratory of Regenerative Medicine,Key Laboratory of Adult Stem Cell Translational Research(Chinese Academy of Medical Sciences),Guizhou Medical University,Guiyang 55025,China;Translational Medicine Research Center,Guizhou Medical University,Guiyang 55025,China)

机构地区：[1]贵州医科大学基础医学院组织学与胚胎学教研室,贵阳550025 [2]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,贵州省再生医学重点实验室,中国医学科学院成体干细胞转化研究重点实验室,贵阳550025 [3]贵州医科大学转化医学研究中心,贵阳550025

出　　处：《解剖学报》2022年第4期453-460,共8页Acta Anatomica Sinica

基　　金：国家自然科学基金(82060151);贵州省科技支撑计划(黔科合支撑〔2020〕4Y230号);贵州医科大学学术新苗项目(19NSP037);中国医学科学院中央级公益性科研院所基本科研业务费项目(2018PT31048,2019PT310013)。

摘　　要：目的探讨NOD/Ltj小鼠自发1型糖尿病(T1D)过程中,胸腺内Wnt信号通路、自身免疫调节因子(AIRE)及T1D组织特异性抗原(TSAs)胰岛素2(Ins2)、谷氨酸脱羧酶67(GAD67)表达与T1D发生的关系。方法60只雌性NOD/Ltj小鼠分3组:3周组、16周未发病组及16周发病组,每组20只;非空腹血糖值连续2次≥11.1 mmol/L视为发生T1D。胰腺HE染色观察胰岛炎发生情况,抗Ins、CD45免疫组织化学染色显示胰岛β细胞或浸润炎性细胞;Western blotting和Real-time PCR检测胸腺Wnt7a、β-catenin、AIRE、Ins、GAD67蛋白水平和mRNA表达;流式细胞术分析胸腺T细胞比例。结果1.随着T1D发生,胰岛组织结构破坏,大量淋巴细胞浸润,残存胰岛细胞减少;Ins^(+)胰岛β细胞周围见大量CD45^(+)细胞聚集。2.随年龄的增加,胸腺Wnt7a、β-连环蛋白(catenin)、AIRE、Ins和GAD67蛋白水平和mRNA表达降低;同周龄发病组小鼠与未发病组小鼠相比,胸腺Ins表达下降。3.与3周组相比,16周未发病组CD4、CD8单阳性T细胞比例降低;16周发病组CD4、CD8单阳性T细胞比例升高,CD4、CD8双阳性T细胞比例降低。结论Wnt7a/β-catenin信号通路可能通过调控AIRE表达,下调T1D相关TSAs表达,参与T1D发生发展。Objective To investigate the relationship between the expression of Wnt signaling pathway,autoimmune regulator(AIRE)and type 1 diabetes(T1D)tissue specific antigen(TSAs)insulin 2(Ins2)and glutamic acid decarboxybase(GAD67)in thymus and the occurrence of T1D in NOD/Ltj mice with spontaneous type 1 diabetes(T1D).Methods Sixty female NOD/Ltj mice were divided into three groups:3 weeks group,16 weeks non-onset group and 16 weeks onset group.Two consecutive non-fasting blood glucose levels≥11.1 mmol/L were considered as the occurrence of T1D.Pancreatic HE staining was used to observe the occurrence of islet inflammation.Anti-Ins and CD45 immunohistochemical staining showed isletβcells or infiltrating inflammatory cells.The protein levels and mRNA expressions of Wnt7a,β-catenin,AIRE,Ins and GAD67 in thymus were detected by Western blotting and Real-time PCR.The proportion of T cells in thymus was analyzed by flow cytometry.Results 1.With the occurrence of T1D,the islet structure was destroyed,a large number of lymphocytes infiltrated,and the remaining islet cells were reduced.A large number of CD45^(+)cells were observed around Ins^(+)isletβcells.2.The protein levels and mRNA expressions of Wnt7a,β-catenin,AIRE,Ins2 and GAD67 in thymus decreased with age.The expression of Ins in thymus decreased in the same week-old group compared with that in the control group.3.Compared with the 3 weeks group,the proportion of CD4 and CD8 single positive T cells in the 16 weeks group was lower;In the 16 weeks group,the proportion of CD4 and CD8 single positive T cells increased,while the proportion of CD4 and CD8 double positive T cells decreased.Conclusion Wnt7a/β-catenin signaling pathway may be involved in the occurrence and development of T1D by regulating AIRE expression and down-regulating T1D-related TSAs expression.

关 键 词：1型糖尿病 自身免疫调节因子 WNT信号通路 免疫印迹法 实时定量聚合酶链反应 NOD/Ltj小鼠 

分 类 号：R725[医药卫生—儿科]