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目的 分析幽门螺杆菌(Hp)感染对自身免疫性甲状腺炎(AIT)小鼠甲状腺损伤与凋亡的影响。方法 51只雌性C57BL/6J小鼠随机分为正常对照组(Control组)、实验性AIT组(EAT组)、Hp感染组(EAT+Hp组),每组17只；除Control组，其余两组小鼠均建立EAT模型；EAT造模成功后，EAT+Hp组小鼠在两周内接受了7次1×109 CFU/mL的Hp菌液灌胃，Control组和EAT组则同步给予等体积生理盐水；末次Hp干预后12周，采用酶联免疫吸附试验(ELISA)检测小鼠血清抗过氧化物酶抗体(TPOAb)、抗甲状腺球蛋白抗体(TGAb)、游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺素(TSH),以及炎症因子白介素-17A(IL-17A)、白介素-10(IL-10)和转化生长因子-β1(TGF-β1)水平；苏木素-伊红(HE)染色评估甲状腺组织病理形态并进行炎症评分；原位末端标记法(TUNEL)检测小鼠甲状腺细胞凋亡情况；实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测甲状腺Fas与Caspase-3的mRNA表达水平；蛋白免疫印迹法(Western blot)和免疫组织化学(IHC)染色检测甲状腺Fas与裂解Caspase-3蛋白表达情况。结果 EAT模型小鼠呈现出甲状腺功能减退特征，血清FT3、FT4水平较Control组显著降低(P<0.05),同时伴有TSH代偿性升高(P<0.01),在此基础上，合并Hp感染进一步加剧了甲状腺功能损伤，FT4进一步下降(P<0.05),TSH水平也相应升高(P<0.05);在自身免疫应答方面，EAT组小鼠TPOAb水平较Control组显著上升(P<0.001),Hp感染促进了自身抗体的进一步生成，TPOAb与TGAb均较EAT组升高(P<0.05);炎症因子检测显示，EAT组促炎因子IL-17A明显高于Control组(P<0.01),而抗炎因子IL-10与TGF-β1则显著降低(P<0.05),Hp感染加剧了这种免疫失衡，使IL-17A进一步上升，IL-10与TGF-β1下降更为明显；组织病理学检查可见EAT组甲状腺结构破坏伴淋巴细胞浸润，Hp感染后损伤程度加重，炎症评分显著升高(P<0.05);在凋亡机制层面，荧光显微镜下见EAT+Hp组较EAT组凋亡阳性细胞表达增多，EAT组甲状腺组织中Fas与Caspase-3 mRNA表达均显著上调(P<0.001),Hp感染后Fas表达进一步升高(P<0.001);Western blot与免疫组织化学结果表明Hp感染促进Fas与裂解Caspase-3蛋白的表达，增强甲状腺细胞凋亡。结论 Hp感染可能破坏Th17/Treg平衡，加剧甲状腺细胞凋亡，从而进一步加重小鼠的AIT。
Abstract：Objective To analyze the impact of Helicobacter pylori(Hp) infection on thyroid injury and apoptosis in a mouse model of autoimmune thyroiditis(AIT). Methods Fifty-one female C57BL/6J mice were randomized into the Control, AIT(EAT), and Hp-infected AIT(EAT+Hp) groups(n=17 in each group). The EAT model was established in the latter two groups. Following successful modeling, EAT+Hp mice received 7 oral gavages of 1×109 CFU/mL Hp over 2 weeks, while the Control group received saline. Twelve weeks after the final intervention, serum levels of TPOAb, TGAb, FT3, FT4, TSH, IL-17A, IL-10, and TGF-β1 were measured via ELISA. Thyroid histopathology(HE staining, inflammation scoring) and apoptosis(TUNEL assay) were evaluated. The mRNA expression of Fas and Caspase-3 was quantified by RT-qPCR, Fas and Cleaved Caspase-3 protein expression was detected by Western blot and immunohistochemistry(IHC). Results Compared with the Control group, EAT mice developed hypothyroidism, characterized by significantly decreased FT3 and FT4 levels and increased TSH(P<0.05). Hp infection worsened this dysfunction, leading to a further reduction in FT4 and a rise in TSH(P<0.05). TPOAb levels were elevated in EAT mice(P<0.001), and Hp infection prompted additional increases in both TPOAb and TGAb(P<0.05). A pro-inflammatory imbalance was observed in EAT mice, with increased IL-17A and decreased IL-10 and TGF-β1(P<0.05). Hp infection aggravated this shift. Histopathological analysis showed more severe thyroid structural damage, lymphocyte infiltration, and higher inflammation scores in the EAT+Hp group(P<0.05). Apoptosis was enhanced in EAT+Hp mice, accompanied by upregulated mRNA expression of Fas and Caspase-3(P<0.001) and increased protein expression of Fas and cleaved Caspase-3. Conclusion Hp infection may exacerbate AIT by disrupting the Th17/Treg balance and promoting thyroid cell apoptosis.
[1] CATUREGLI P,DE REMIGIS A,ROSE N R.Hashimoto thyroiditis:clinical and diagnostic criteria[J].Autoimmun Rev,2014,13(4):391-397.
[2] 迪丽达尔·依马本.幽门螺旋杆菌感染与桥本甲状腺炎的相关性研究[D].乌鲁木齐：新疆医科大学，2024.
[3] 李慧芳，黄金莲，梅起化，等.桥本甲状腺炎与幽门螺旋杆菌感染的相关研究[J].浙江临床医学，2018,20(12):1983-1984.
[4] 马晓媛.根除幽门螺旋杆菌感染对甲状腺自身抗体及功能影响的随访研究[D].乌鲁木齐：新疆医科大学，2023.
[5] 马雅霞，杜学芹，雒芳玲，等.女性桥本甲状腺炎发病与幽门螺旋杆菌感染相关性研究[J].吉林医学，2020,41(9):2104-2106.
[6] CELLINI M,SANTAGUIDA M G,VIRILI C,et al.Hashimoto′s thyroiditis and autoimmune gastritis[J].Front Endocrinol,2017,31(7):892.
[7] CHOI Y M,KIM T Y,KIM E Y,et al.Association between thyroid autoimmunity and Helicobacter pylori infection[J].Korean J Intern Med,2017,32(2):309-313.
[8] FIGURA N,DI CAIRANO G,MORETTI E,et al.Helicobacter pylori infection and autoimmune thyroid diseases:the role of virulent strains[J].Antibiotics (Basel),2019,9(1):12.
[9] HOU Y,SUN W,ZHANG C,et al.Meta-analysis of the correlation between Helicobacter pylori infection and autoimmune thyroid diseases[J].Oncotarget,2017,8(70):115691-115700.
[10]WANG L,CAO Z M,ZHANG L L,et al.Helicobacter pylori and autoimmune diseases:involving multiple systems[J].Front Immunol,2022,13:833424.
[11]ALI A,ALHUSSAINI K I.Helicobacter pylori:A Contemporary perspective on pathogenesis,diagnosis and treatment strategies[J].Microorganisms,2024,12(1):222.
[12]AZIZ F,KHAN I,SHUKLA S,et al.Partners in crime:The Lewis Y antigen and fucosyltransferase IV in Helicobacter pylori-induced gastric cancer[J].Pharmacol Ther,2021,232:107994.
[13]HEYDARZADEH S,ABOOSHAHAB R,ZARKESH M,et al.Endocrine polyautoimmunity:mechanistic insights and the future of AI-driven diagnostics[J].EXCLI journal,2025,24:1500- 1519.
[14]DORE M P,FANCIULLI G,MANCA A,et al.Association of Helicobacter pylori infection with autoimmune thyroid disease in the female sex[J].J Clin Med,2023,12(15):5150.
[15]BEDULEVA L,SIDOROV A,TERENTIEV A,et al.Reduction in experimental autoimmune thyroiditis by IgG Fc fragments bearing regRF epitopes[J].Immunol Res,2023,71(1):83-91.
[16]HE C,LI Y,GAN L,et al.Notch signaling regulates Th17 cells differentiation through PI3K/AKT/mTORC1 pathway and involves in the thyroid injury of autoimmune thyroiditis[J].J Endocrinol Invest,2024,47(8):1971-1986.
[17]LIU H,LI Y,ZHU Y,et al.Notch signaling pathway promotes Th17 cell differentiation and participates in thyroid autoimmune injury in experimental autoimmune thyroiditis mice[J].Mediators Inflamm,2023,2023:1195149.
[18]孙伯菊，吴丽丽，秦灵灵，等.桥本氏甲状腺炎实验模型构建研究进展[J].世界科学技术-中医药现代化，2020,22(7):2205-2210.
[19]WU D,CAO M,LI N,et al.Effect of trimethylamine N-oxide on inflammation and the gut microbiota in Helicobacter pylori-infected mice[J].Int Immunopharmacol,2020,81:106026.
[20]WU D,CAO M,PENG J,et al.The effect of trimethylamine N-oxide on Helicobacter pylori-induced changes of immunoinflammatory genes expression in gastric epithelial cells[J].Int Immunopharmacol,2017,43:172-178.
[21]VERGINIS P,LI H S,CARAYANNIOTIS G.Tolerogenic semimature dendritic cells suppress experimental autoimmune thyroiditis by activation of thyroglobulin-specific CD4+CD25+T cells[J].J Immunol,2005,174(11):7433-7439.
[22]WARREN J R,MARSHALL B.Unidentified curved bacilli on gastric epithelium in active chronic gastritis[J].Lancet,1983,1(8336):1273-1275.
[23]DONIACH D,RO ITT I M,TAYLOR K B.Autoimmune phenomena in pernicious anaemia.Serological overlap with thyroiditis,thyrotoxicosis,and systemic lupus erythematosus[J].Br Med J,1963,1(5342):1374-1379.
[24]CENTANNI M,MARIGNANI M,GARGANO L,et al.Atrophic body gastritis in patients with autoimmune thyroid disease:an underdiagnosed association[J].Arch Intern Med,1999,159(15):1726-1730.
[25]MORAN A P,PRENDERGAST M M,APPELMELK B J.Molecular mimicry of host structures by bacterial lipopolysaccharides and its contribution to disease[J].FEMS Immunol Med Microbiol,1996,16(2):105-115.
[26]TOMASI P A,DORE M P,FANCIULLI G,et al.Is there anything to the reported association between Helicobacter pylori infection and autoimmune thyroiditis?[J].Dig Dis Sci,2005,50(2):385-388.
[27]HE H,JIANG Y,QIU J,et al.Role of interleukin 17 and T helper cells 17 cells as a new immune target and signalling in the pathogenesis and treatment of autoimmune thyroid diseases[J].Ann Med,2025,57(1):2586216.
[28]BETTELLI E,CARRIER Y,GAO W,et al.Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells[J].Nature,2006,441(7090):235-238.
[29]SAKAGUCHI S,MIKAMI N,WING J B,et al.Regulatory T cells and human disease[J].Annu Rev Immunol,2020,38:541-566.
[30]ANDRIKOULA M,TSATSOULIS A.The role of Fas-mediated apoptosis in thyroid disease[J].Eur J Endocrinol,2001,144(6):561-568.
[31]SALMASO C,BAGNASCO M,PESCE G,et al.Regulation of apoptosis in endocrine autoimmunity:insights from Hashimoto′s thyroiditis and Graves′ disease[J].Ann N Y Acad Sci,2002,966:496-501.
基本信息:
DOI：10.19367/j.cnki.2096-8388.2026.03.003
中图分类号:R581
引用信息:
[1]王才虹,华宇羽,覃丹,等.幽门螺杆菌感染促进EAT小鼠甲状腺损伤与凋亡的实验研究[J].贵州医科大学学报,2026,51(03):333-342.DOI:10.19367/j.cnki.2096-8388.2026.03.003.
基金信息:
国家自然科学基金项目(82260160)
2026-03-20
2026-03-20