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实验性牙周炎大鼠牙龈组织及外周血中Th17/Treg细胞及相关因子的检测

Detection of Th17/Treg cells and related factors in gingival tissues and peripheral blood of rats with experimental periodontitis.

作者信息

Gao Li, Zhao Yajing, Wang Panpan, Zhang Liping, Zhang Chi, Chen Qianying, Zhao Chuanjiang

机构信息

Department of Periodontology, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat -sen University, Guangzhou 510055, China.

Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China.

出版信息

Iran J Basic Med Sci. 2017 Mar;20(3):294-300. doi: 10.22038/ijbms.2017.8359.

Abstract

OBJECTIVES

This study aimed to investigate the role and the possible mechanisms involved in the immunoregulation of experimental periodontitis by Th17/Treg.

MATERIALS AND METHODS

Experimental periodontitis was established by silk thread ligation with daubing in the bilateral maxillary second molar of Male Sprague-Dawley (SD) rats. Alveolar bones were scanned by Micro-CT. Histological examination was stained with H&E. The proportions of Th17 and Treg cells in peripheral blood were detected by flow cytometry. RT-PCR was used to measure the expression of RORγt, Foxp3 mRNA in the gingival tissues. The concentrations of IL-17, IL-10, and TGF-β in peripheral blood and gingival crevicular fluid were measured by ELISA.

RESULTS

Experimental rats showed profound bone resorption and inflammatory cell infiltration. The percentages of Th17 significantly increased in the peripheral blood, which was consistent with gingival tissues study that Th17 cells related transcription factor RORγt mRNA and IL-17 increased in the course of periodontitis. The percentages of CD25Foxp3 Treg significantly increased in the peripheral blood, which was consistent with gingival tissues study that Treg cells related transcription factor Foxp3 mRNA and cytokines IL-10 and TGF-β increased in the course of periodontitis. The ratio of Th17/Treg cells was significantly increased in the peripheral circulation, however, the Th17/Treg balance is in wave motion in inflamed gingival tissues in the different stages of periodontitis.

CONCLUSION

Th17/Treg balance may be associated with the progression of periodontitis and pathological tissue destruction. Moreover, local inflammation would result in the up-regulation ratio of Th17/Treg in peripheral blood, which may influence some periodontally involved systemic diseases.

摘要

目的

本研究旨在探讨Th17/Treg在实验性牙周炎免疫调节中的作用及可能机制。

材料与方法

采用丝线结扎并涂抹于雄性Sprague-Dawley(SD)大鼠双侧上颌第二磨牙的方法建立实验性牙周炎。通过Micro-CT扫描牙槽骨。组织学检查采用苏木精-伊红(H&E)染色。采用流式细胞术检测外周血中Th17和Treg细胞的比例。运用逆转录聚合酶链反应(RT-PCR)检测牙龈组织中RORγt、Foxp3 mRNA的表达。采用酶联免疫吸附测定(ELISA)法检测外周血和龈沟液中白细胞介素-17(IL-17)、白细胞介素-10(IL-10)和转化生长因子-β(TGF-β)的浓度。

结果

实验大鼠出现明显的骨吸收和炎性细胞浸润。外周血中Th17的百分比显著增加,这与牙龈组织研究结果一致,即牙周炎过程中Th17细胞相关转录因子RORγt mRNA和IL-17增加。外周血中CD25Foxp3 Treg的百分比显著增加,这与牙龈组织研究结果一致,即牙周炎过程中Treg细胞相关转录因子Foxp3 mRNA以及细胞因子IL-10和TGF-β增加。外周循环中Th17/Treg细胞的比例显著增加,然而,在牙周炎不同阶段的炎症牙龈组织中,Th17/Treg平衡呈波动状态。

结论

Th17/Treg平衡可能与牙周炎的进展及病理组织破坏有关。此外,局部炎症会导致外周血中Th17/Treg比例上调,这可能会影响一些与牙周炎相关的全身性疾病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b765/5378967/91994d469944/IJBMS-20-294-g001.jpg

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