Treg/Th17、CD4+/CD8+ T淋巴细胞在实验性牙周炎大鼠中的表达及意义

doi:10.19439/j.sjos.2022.04.007

上海口腔医学 ›› 2022, Vol. 31 ›› Issue (4): 374-378.doi: 10.19439/j.sjos.2022.04.007

Treg/Th17、CD4⁺/CD8⁺ T淋巴细胞在实验性牙周炎大鼠中的表达及意义

莫鹏¹, 李筱玥², 刘耀徽³

1.青海省人民医院口腔科, 2病理科,青海西宁 810000;
3.桂林市口腔医院牙体牙髓病科,广西桂林 541000

收稿日期:2021-10-08 修回日期:2021-11-19 出版日期:2022-08-25 发布日期:2022-08-30
通讯作者: 莫鹏,E-mail: 13897240309@163.co
作者简介:莫鹏（1987-）,男,本科,主治医师
基金资助:
青海省科技厅自然科学基金（2020-ZJ-963Q）

Expression and significance of Treg/Th17 and CD4⁺/CD8⁺ T lymphocytes in experimental periodontitis in rats

MO Peng¹, LI Xiao-yue², LIU Yao-hui³

1. Department of Stomatology, 2. Department of Pathology, Qinghai Provincial People's Hospital. Xining 810000, Qinghai Province;
3. Department of Endodontics, Guilin Stomatological Hospital. Guilin 541000, Guangxi Zhuang Autonomous Region, China

Received:2021-10-08 Revised:2021-11-19 Online:2022-08-25 Published:2022-08-30

摘要/Abstract

摘要： 目的: 探讨辅助性T细胞17/调节性T细胞（Th17/Treg）、CD4⁺/CD8⁺ T淋巴细胞在实验性牙周炎大鼠中的表达,并分析其临床意义。方法: 取20只SPF级SD雄性大鼠,随机分为牙周炎组（将牙龈卟啉单胞菌混悬液接种至牙龈沟内）和对照组,每组10只。实验组术后1周内,隔天以牙龈卟啉单胞菌混悬液涂抹大鼠口腔,2组分笼饲养8周。麻醉处死后,取大鼠左侧上颌第二磨牙颌骨组织,以亚甲基蓝染色观察并测定牙槽骨丧失（ABL）情况;H-E染色观察颌骨组织病理学变化。分离并培养大鼠外周血单个核细胞（PBMC）及T细胞;流式细胞术检测外周血中Treg、Th17细胞及CD4⁺、CD8⁺ T淋巴细胞,酶联免疫吸附（ELISA）法检测血清白介素17（IL-17）、IL-10、IL-4和干扰素γ（INF-γ）,免疫印迹法检测颌骨组织中维甲酸相关孤儿核受体（RORγt）、叉头状翅膀样转录因子3（FOXP3）和缝隙连接蛋白（Cx40）蛋白的表达变化。采用SPSS 19.0软件包对数据进行统计学分析。结果: 与对照组相比,牙周炎组大鼠ABL、外周血Th17比例、Th17/Treg比值、CD4⁺比例、CD4⁺/CD8⁺ T淋巴细胞比值、血清IL-17、IL-10及IL-4、颌骨组织中Foxp3及Cx40蛋白显著升高（P<0.05）,Treg比例、INF-γ、颌骨组织中RORγt蛋白显著降低（P<0.05）。结论: Treg/Th17、CD4⁺/CD8⁺ T淋巴细胞平衡失衡,造成炎症因子IL-17、IL-10及IL-4异常高表达,可能与实验性牙周炎发病关系密切,为有效治疗牙周炎提供了理论依据。

关键词: 实验性牙周炎, 辅助性T细胞17/调节性T细胞, CD4⁺/CD8⁺ T淋巴细胞

Abstract: PURPOSE: To explore the expression of helper T cell 17/regulatory T cell (Th17/Treg) and CD4⁺/CD8⁺ T lymphocyte in experimental periodontitis in rats, and analyze its clinical significance. METHODS: Twenty SPF male SD rats were randomly divided into experimental group (inoculate Porphyromonas gingivalis suspension into gingival sulcus) and control group, with 10 rats in each group. The experimental group was smeared with Porphyromonas gingivalis suspension every other day within 1 week after operation, and the two groups were caged for 8 weeks. After the rats were sacrifical under anesthesia, the jaw tissue of the left maxillary second molar was stained with methylene blue to observe and measure the loss of alveolar bone (ABL). Hematoxylin-eosin (H-E) staining was used to observe the histopathological changes of the jaw. Rat peripheral blood mononuclear cells (PBMC) and T cells were isolated and cultured, Treg, Th17 cells and CD4⁺, CD8⁺ T lymphocytes in peripheral blood were detected by flow cytometry. The levels of serum interleukin-17(IL-17), IL-10 and IL-4, INF-γ were detected by enzyme-linked immunosorbent assay (ELISA). Expression changes of retinoic acid related orphan nuclear receptor (RORγt), forkhead wing like transcription factor 3 (Foxp3) and gap junction protein(Cx40) in jaw tissue were detected by Western blot. The data were statistically analyzed by SPSS 19.0 software package. RESULTS: Compared with the control group, ABL, peripheral blood Th17 ratio, Th17/Treg ratio, CD4⁺ratio, CD4⁺/CD8⁺ T lymphocyte ratio, serum IL-17, IL-10 and IL-4 level, Foxp3 and Cx40 protein in jaw tissue were signifinantly increased in the experimental group(P<0.05), while Treg ratio, INF-γ, RORγt protein in jaw tissue significantly decreased(P<0.05) in the experimental group. CONCLUSIONS: Imbalance of Treg/Th17 and CD4⁺/CD8⁺ T lymphocytes leads to the abnormal high expression of inflammatory factors IL-17, IL-10 and IL-4, which may be closely related to the pathogenesis of experimental periodontitis.

Key words: Experimental periodontitis, Helper T cells 17/regulatory T cells, CD4⁺/CD8⁺ T lymphocytes

中图分类号:

R781.4

莫鹏, 李筱玥, 刘耀徽. Treg/Th17、CD4⁺/CD8⁺ T淋巴细胞在实验性牙周炎大鼠中的表达及意义[J]. 上海口腔医学, 2022, 31(4): 374-378.

MO Peng, LI Xiao-yue, LIU Yao-hui. Expression and significance of Treg/Th17 and CD4⁺/CD8⁺ T lymphocytes in experimental periodontitis in rats[J]. Shanghai Journal of Stomatology, 2022, 31(4): 374-378.

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Treg/Th17、CD4⁺/CD8⁺ T淋巴细胞在实验性牙周炎大鼠中的表达及意义

Expression and significance of Treg/Th17 and CD4⁺/CD8⁺ T lymphocytes in experimental periodontitis in rats

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Treg/Th17、CD4+/CD8+ T淋巴细胞在实验性牙周炎大鼠中的表达及意义

Expression and significance of Treg/Th17 and CD4+/CD8+ T lymphocytes in experimental periodontitis in rats

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Treg/Th17、CD4⁺/CD8⁺ T淋巴细胞在实验性牙周炎大鼠中的表达及意义

Expression and significance of Treg/Th17 and CD4⁺/CD8⁺ T lymphocytes in experimental periodontitis in rats