【作者】 石铁； 林晓萍；

【机构】 中国医科大学盛京医院；

【摘要】 目的 :以牙周炎病理模型为分析对象,通过动物实验及细胞实验,采用ELISA、Flow cytometry、实时荧光定量PCR等多种免疫研究技术分析并说明牙周炎对B10细胞的影响。方法 :我们首先建立了牙周炎动物模型。提取模型动物与健康动物的牙龈及外周血中的淋巴细胞,分析不同状态下不同组织内IL-10+B细胞的变化趋势及特点。另取健康的脾源性B细胞,在常规条件下以及在Pg-LPS诱导下培养一周后分别采用流式细胞术、ELISA以及实时荧光定量PCR从基因及蛋白水平评价各培养条件下B细胞表达分泌IL-10的水平。拟揭示牙周炎宿主B10细胞的变化特点并分析牙周致病菌对该细胞分化及功能的影响。结果 :无论是健康个体还是牙周炎宿主,均可在其牙龈及外周血液中检测到B10细胞的存在。在牙周炎时,局部组织内B10细胞比例显著升高,但在外周血中却并未发现上述变化。在牙周炎进程中,B10细胞的迅速升高更多的体现在疾病早期,在病程晚期上此变化趋于平稳。将初始B细胞与牙周致病菌共培养,其内的B10细胞分化水平显著升高,细胞内IL-10 mRNA水平以及培养基中IL-10浓度也同时升高。结论 :牙周致病菌促进宿主B10细胞分化及功能。更多还原

【Abstract】 Objective: In the present study, we set experimental periodontitis models. How periodontopathogen affects B10 cells was established and how B10 cells regulate Th17/Treg during periodontitis were analyzed by ELISA, Flow Cytometry, qPCR, Also, we performed cell culture to furtherly clarify the potential mechanisms of B10 cells on regulating Th17/Treg. Methods: We set an experimental periodontitis model for the present research. The changing characteristics of IL-10 + B cells in different tissues were analyzed. In addition, healthy splenic B cells were cultured for one week independently or with Pg-LPS. The expression of IL-10 was evaluated by flow cytometry, ELISA, and qPCR. Results: B10 cells are detected in gingiva and peripheral blood of both healthy individuals and periodontitis ones. In periodontitis, the proportion of B10 cells in local lesions was significantly increased, but not in peripheral blood. B10 cells presented a rapid increase in the early stage of periodontitis, but a smoothed change in the later stage. When co-cultured with periodontopathogens, the proportion of B10 cells was significantly increased, and the IL-10 mRNA level and the concentration of IL-10 in the medium were also increased. Conclusions: Periodontal pathogens promote the differentiation and function of B10 cells.更多还原