Zeste同源物2增强子在桥本甲状腺炎B淋巴细胞亚群中的表达及其抑制剂的治疗机制及效果研究  

EZH2 Expression in B Cell Lymphocyte Subsets of Hashimoto's Thyroiditis and the Therapeutic Mechanism and Effect of Its Inhibitors

作　　者：易圣果 曹业迪 赵雪[1] 卢桂芝[1] 张杨[1] 丛铁川[2] 张澜波[3] 张继新[4] 梁振威 屈晨雪[6] 张俊清[1] 高莹[1] YI Shengguo;CAO Yedi;ZHAO Xue;LU Guizhi;ZHANG Yang;CONG Tiechuan;ZHANG Lanbo;ZHANG Jixin;LIANG Zhenwei;QU Chenxue;ZHANG Junqing;GAO Ying(Department of Endocrinology,Peking University First Hospital,Beijing 100034,China;Department of Otorhinolaryngology-Head and Neck Surgery,Peking University First Hospital,Beijing 100034,China;Department of Thyroid and Breast Surgery,Peking University First Hospital,Beijing 100034,China;Department of Pathology,Peking University First Hospital,Beijing 100034,China;Department of Ultrasound Medicine,Peking University First Hospital,Beijing 100034,China;Department of Clinical Laboratory,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院内分泌内科,北京市100034 [2]北京大学第一医院耳鼻咽喉-头颈外科,北京市100034 [3]北京大学第一医院甲状腺乳腺外科,北京市100034 [4]北京大学第一医院病理科,北京市100034 [5]北京大学第一医院超声医学科,北京市100034 [6]北京大学第一医院检验科,北京市100034

出　　处：《中国全科医学》2024年第21期2639-2645,共7页Chinese General Practice

基　　金：国家自然科学基金面上项目(8217030553)。

摘　　要：背景甲状腺自身抗体是诊断桥本甲状腺炎(HT)的标志,B淋巴细胞在HT的发病机制中发挥重要作用。Zeste同源物2增强子(EZH2)是一种表观遗传学蛋白,在淋巴细胞的发育与功能调控中扮演重要角色。目的本研究探讨EZH2在HT甲状腺组浆母细胞及浆细胞中的表达,进一步探讨EZH2抑制剂在实验性自身免疫甲状腺炎(EAT)模型中的治疗作用。方法收集北京大学第一医院2010—2020年6例行甲状腺手术的患者,取肿瘤对侧的甲状腺组织(HT及正常甲状腺组织各3例),通过RNA-seq筛选B淋巴细胞相关基因的表达情况;收集16例HT患者的甲状腺组织和8例健康对照(HD)甲状腺组织,分别利用免疫组化及免疫荧光验证EZH2在HT甲状腺组织中B淋巴细胞中的表达;收集25例HT甲状腺细针穿刺液(FNA)、19例HT外周血以及12例健康人外周血样本应用流式细胞分析检测EZH2在浆母细胞及浆细胞中的表达改变。将15只7周龄NOD.H-2^(h4)小鼠EAT模型分为对照组(n=5)、EAT无注射(n=5)或注射EZH2抑制剂GSK126处理组(10 mg/kg,腹腔注射3次/周,n=5),8周后观察甲状腺炎症程度及甲状腺球蛋白抗体(TgAb)水平的改变。结果RNA-seq结果显示,相较于正常甲状腺组织,HT甲状腺组织中EZH2水平上调,一些与B淋巴细胞表型相关的基因例如CD19、CD27、CD38、CD52相应增加。免疫组化结果显示,16例HT甲状腺组织标本中EZH2免疫组化染色均可在生发中心(GC)见到阳性细胞,呈强阳性,8例正常甲状腺组织染色中未观察到阳性细胞。HT甲状腺组织中EZH2染色高表达在GC区,EZH2特异性地表达在CD_(19)^(+)B淋巴细胞中。流式细胞术检测结果显示HT FNA样本中CD_(19)^(+)B淋巴细胞、浆母细胞及浆细胞比例均高于HD外周血、HT外周血样本(P<0.01),HT FNA样本中EZH2在CD_(19)^(+)B淋巴细胞、浆母细胞中的阳性比例高于HT外周血(P<0.005)。小鼠实验中,EAT组甲状腺的淋巴细胞浸润较对照组增加。GSK1Background Thyroid autoantibody is a marker for the diagnosis of Hashimoto's thyroiditis(HT),and B cells are essential in the pathogenesis of HT.Enhancer of Zeste homolog 2(EZH2),which is an important epigenetic regulator,plays an important role in the regulation of lymphocytes development and function.Objective To investigate EZH2 expression in plasmoblasts and plasma cells in HT,and further explore the therapeutic effect of EZH2 inhibitors in experimental autoimmune thyroiditis(EAT) model.Methods The thyroid tissues from 6 patients who underwent thyroidectomy(3 HT patients with PTC,3 patients with PTC alone) in Peking University First Hospital between 2010 and 2020 were obtained from the contralateral lobe with thyroid cancer,and screened for the expression of B-lymphocyte-related genes by RNA-seq;thyroid tissues from 16 HT patients and 8 normal thyroid tissues were collected and verified for the the expression of EZH2 in B cells in HT thyroid tissues by immunohistochemistry or immunofluorescence,respectively.Fine-needle aspiration(FNA) samples from patients with HT(n=25),and peripheral blood from patients with HT(n=19) or healthy donors(n=12) were analyzed by flow cytometry to define altered EZH2 expression in plasmablasts and plasma cells.Fifteen seven-week-old NOD.H-2^(h4) mice were randomly divided into the control(n=5),EAT without injection group(n=5),and EZH2 inhibitor+ GSK126 injection group(10 mg/kg,intraperitoneal injection 3 times/week,n=5).The degree of thyroid inflammation and changes in TgAb levels were observed after 8 weeks.Results RNA sequencing analysis showed that EZH2 and genes associated with the B-cell phenotype such as CD19,CD27,CD38,CD52 were higher expressed in HT hyroid tissues compared with normal thyroid tissues.Immunohistochemical results showed that immunohistochemical staining for EZH2 in 16 HT thyroid tissue specimens was strongly positive with positive cells observed in the GC region,and no positive cells were observed in the staining of 8 normal thyroid tissues.EZH2 staining in

关 键 词：桥本甲状腺炎 B淋巴细胞亚群 Zeste同源蛋白2增强子 甲状腺功能减退症 靶向治疗 

分 类 号：R581.4[医药卫生—内分泌]