实验性牙周炎中调节性T细胞的检测及免疫状态

Detection and the immune status of regulatory T cells in experimental periodontitis

目的:通过口腔涂抹牙周致病菌加腹腔注射抗原的方法建立一种模拟人类牙周病发病的动物模型,分析调节性T细胞在牙周炎小鼠中的表达情况。方法:采用实体显微镜评价牙周骨吸收的程度,HE染色法观察牙周组织病变的炎细胞浸润情况;FACS检测调节性T细胞在牙周炎慢性炎症期间的表达;实时定量PCR检测牙龈组织中TGF-β1、IL-10mRNA的表达。采用SPSS 17.0软件包对数据进行统计学分析。结果:第4周后,牙周炎组与正常对照组相比,釉牙骨质界至牙槽嵴顶的距离显著增加(P<0.05);牙周组织中大量的炎性细胞浸润,牙周袋加深;TGF-β1、IL-10mRNA的表达水平显著降低(P<0.01);脾脏中调节性T细胞的比率及数量显著少于正常对照组(P<0.05)。结论:牙周炎小鼠中调节性T细胞的数量及功能存在异常,可能与其分泌的免疫抑制性细胞因子表达降低有关。

PURPOSE： To establish animal model in mice with orally infected periodontitis with Porphyromonas gingivalis and intraperitoneal injection of P.g antigen, simulate periodontal pathogens of human periodontal disease, and analyze the expression of regulatory T cells in this model. METHODS： The extent of periodontal bone resorption was measured by entity microscope, and inflammatory cell infiltration of the periodontal tissue lesions was observed by HE staining. Flow cytometric analysis was used to detect the expression of regulatory T cells in periodontal disease during the chronic inflammation. The expression levels of TGF-β1, IL-10mRNA in periodontal tissue were detected by real-time quantitative PCR. SPSS 17.0 software package was used for statistical analysis. RESULTS： After 4 weeks, compared with the normal control group, the distance between cemento-enamel junction and the alveolar crest was significantly increased in periodontal disease group （P〈0.05）; Massive inflammatory cells were infiltrated and periodontal pockets deepened in periodontal tissue. The expression levels of TGF-IS1 and IL-10mRNA were significantly decreased （P〈0.01）. The ratio and numbers of Treg cells were significantly reduced in the spleen （P〈0.05）. CONCLUSIONS： In experimental periodontitis, the number and the function of regulatory T cells are abnormal. It may be relevant to the reduction of immune suppression cytokines. Supported by Higher Institute School Special Research Fund for Doctorate Program （20112104110013） and Science and Technology Project of Liaoninz Province （2012225015）.