Notch1信号通路调控γδT17细胞表达参与EAT小鼠甲状腺自身免疫损伤  

The Notch1 signaling pathway is involved in thyroid autoimmune injury in EAT mice by regulating γδT17 cells

作　　者：刘昊 李依雯 朱玉娇 李玥 薛海波[1] LIU Hao;LI Yi-wen;ZHU Yu-jiao;LI Yue;XUE Hai-bo(Department of Endocrinology,Binzhou Medical University Hospital,Binzhou 256603,China)

机构地区：[1]滨州医学院附属医院内分泌科,滨州256603

出　　处：《现代免疫学》2023年第4期289-295,共7页Current Immunology

基　　金：山东省重点研发计划(2016GSF201021);山东省高等学校科技计划(J16LL01);山东省研究生教育创新计划项目(20038612);滨州医学院附属医院科研创新团队(202031);滨州医学院附属医院后备领军人才(JC2019-03)。

摘　　要：为探讨γ-分泌酶抑制剂阻断Notch1信号通路对实验性自身免疫性甲状腺炎(experimental autoimmune thyroiditis,EAT)小鼠γδT17细胞表达及甲状腺自身免疫损伤的影响,将30只雌性C57BL/6小鼠随机分为正常对照(normal control,NC)组(n=10)、EAT-A组[给予猪甲状腺免疫球蛋白(porcine thyroid immunoglobulin,pTg)多点皮下注射,n=10]、EAT-B组{pTg皮下注射前给予γ-分泌酶抑制剂3,5-二氟苯乙酰-L-丙氨酰-S-苯基甘氨酸t-丁酯[N-(N-3,5-difluorophenacetyl-L-alanyl)-S-phenylglycine t-butyl ester,DAPT]腹腔注射,n=10}。应用H-E染色、ELISA、流式细胞术、qRT-PCR等评估甲状腺炎症程度,检测γδT17细胞比例及其效应细胞因子IL-17A、Notch1信号通路主要组分的表达水平。结果显示,EAT-A组小鼠血清甲状腺球蛋白抗体(thyroglobulin antibody,TgAb)滴度,IL-17A水平,γδT17细胞比例,Notch1、发状分裂相关增强子1(hairy and enhancer of split1,Hes1)及IL-17A mRNA表达水平均显著高于NC组(P<0.01)。EAT-B组小鼠经DAPT处理后上述指标均显著下降(P<0.01),伴有甲状腺内淋巴细胞浸润程度减轻。EAT小鼠γδT17细胞比例与血清TgAb滴度及IL-17A水平均呈正相关(r值分别为0.598、0.497,P<0.05)。该研究提示,γ-分泌酶抑制剂可能通过阻断Notch1信号通路下调EAT小鼠γδT17细胞表达,减轻甲状腺自身免疫损伤。The purpose of this study is to investigate the effect ofγ-secretase inhibitor in regulatingγδT17 cells by blocking the Notch1 signaling pathway on thyroid autoimmune injury in experimental autoimmune thyroiditis(EAT)mice.Thirty female C57BL/6 mice were randomly divided into normal control(NC)group(n=10),EAT-A group(treated with porcine thyroid immunoglobulin[pTg],n=10),and EAT-B group(treated with N-[N-3,5-difluorophenacetyl-L-alanyl]-S-phenylglycine t-butyl ester[DAPT]before pTg,n=10).H-E staining,ELISA,flow cytometry,and qRT-PCR were used to assess the degree of thyroiditis and detect the expressions ofγδT17 cells and their effector cytokine IL-17A,the main components of Notch1 signaling pathway,respectively.The results showed that the serum titre of thyroglobulin antibody(TgAb),the level of IL-17A,the proportion ofγδT17 cells,and the Notch1,hairy and enhancer of split 1(Hes1),IL-17A mRNA expressions in the EAT-A group were all significantly higher compared to those in the NC group(P<0.01).In contrast,in the EAT-B group,the above indexes were all significantly decreased upon DAPT treatment(P<0.01),accompanied by reduced lymphocyte infiltration in the thyroid.Additionally,the proportion ofγδT17 cell in EAT mice was positively correlated with the serum concentrations of TgAb and IL-17A(r=0.598,0.497,P<0.05).Therefore,the results suggest thatγ-secretase inhibitor can reduce thyroid autoimmune injury by blocking the Notch1 signaling pathway to downregulate theγδT17 cells in EAT mice.

关 键 词：γδT17细胞 自身免疫性甲状腺炎 Γ-分泌酶抑制剂 NOTCH1 白细胞介素17 

分 类 号：R392.6[医药卫生—免疫学]