摘要： 目的 采用抗体中和方法抑制调节性T细胞（Treg）的功能，研究Treg细胞在慢性牙周炎小鼠中的作用。方法 通过牙龈卟啉单胞菌（P. gingivalis）感染口腔建立牙周炎小鼠模型。将16只C57BL/6小鼠随机分为Sham组、P. gingivalis组、IgG组、anti-GITR组，每组4只。anti-GITR组、IgG组分别腹腔注射抗小鼠GITR单克隆抗体和同型对照抗体IgG。测量釉牙骨质界到牙槽嵴顶的距离，评价牙槽骨的吸收情况；HE染色观察牙周组织中附着丧失和炎性浸润情况；流式细胞术检测牙龈中CD4⁺Foxp3⁺（Treg）细胞比例和细胞数量；实时PCR检测牙龈中Treg细胞相关因子白细胞介素-10（IL-10）、转化生长因子-β1（TGF-β1）、叉头翼螺旋转录分子（Foxp3）的mRNA表达。结果 与P. gingivalis组和IgG组相比，anti-GITR组小鼠釉牙骨质界到牙槽嵴顶距离增大（P<0.05），牙龈中Treg细胞比例和细胞数量降低（P<0.05）；Treg细胞相关因子IL-10、TGF-β1、Foxp3 mRNA的表达明显下调（P<0.05）。结论 抑制Treg细胞的功能后，小鼠牙周组织破坏加重，表明Treg细胞介导的免疫应答参与慢性牙周炎的炎症调控和组织修复。

调节性T细胞在慢性牙周炎小鼠模型中的作用

张文娟¹, 谭昭¹, 蔡如佳¹, 关宁², 高秀秋¹, 王琳源¹

1. 锦州医科大学附属第二医院牙周黏膜科, 辽宁 锦州 121000;
2. 锦州医科大学附属第一医院脑与脊髓损伤重点实验室, 辽宁 锦州 121000

收稿日期:2020-04-27出版日期:2021-02-28发布日期:2021-01-21
通讯作者:王琳源E-mail:wanglinyuan2006@163.com
作者简介:张文娟(1995-),女,硕士研究生.
基金资助:国家自然科学基金青年基金（81600870）；辽宁省博士启动基金（201601360）


关键词: 慢性牙周炎, 调节性T细胞, 糖皮质激素诱导的肿瘤坏死因子受体
Abstract: Objective To investigate the role of regulatory T (Treg) cells in mice with chronic periodontitis by inhibiting the function of Treg cells using an antibody neutralization method. Methods Sixteen C57BL/6 mice were randomly assigned to a Sham group,a Porphyromonas gingivalis (P. gingivalis) group,an IgG group,and an anti-GITR group (n=4 per group). Experimental periodontitis was induced by oral infection with P. gingivalis. Mice in anti-GITR and IgG groups were intraperitoneally injected with InVivoMAb anti-mouse GITR and InVivoMAb rat IgG2b isotype control,respectively. Alveolar bone resorption was estimated by measuring the distance between the cemento-enamel junction (CEJ) and the alveolar bone crest (ABC). Hematoxylin-eosin staining was performed to examine inflammation and destruction of periodontal tissues. CD4⁺ Foxp3⁺ (Treg) cells were analyzed using flow cytometry. The mRNA of Treg cell-related cytokines IL-10,TGF-β1,and Foxp3 was quantified using real-time PCR. Results The CEJ-ABC distance increased in the anti-GITR group compared with the P. gingivalis and IgG groups. The proportion of CD4⁺ Foxp3⁺ (Treg) cells among total CD4⁺ T cells and abundance of CD4⁺ Foxp3⁺ (Treg) cells in gingival tissue decreased,and expression of Treg cell-related cytokines IL-10,TGF-β1,and Foxp3 was significantly down-regulated (P<0.05). Conclusion The periodontal tissue damage in mice was aggravated by the inhibition of Treg cell function,indicating that immune responses mediated by Treg cells are involved in the regulation of inflammation and tissue repair in chronic periodontitis.
Key words: chronic periodontitis, regulatory T cell, glucocorticoid-induced tumor necrosis factor receptor
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