Cao G Q, Zhang X, Zhao Y, Zhao S Q, Dong G C, Gao Q X, Wang Z M, Lin J
Department of Stomatology, The Fourth Hospital of Harbin Medical University, Harbin 150001, China (is working on the Department of Stomatology, Jingzhou First People's Hospital, Jingzhou 434100, China).
Department of Stomatology, The Fourth Hospital of Harbin Medical University, Harbin 150001, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2019 Aug 9;54(8):553-560. doi: 10.3760/cma.j.issn.1002-0098.2019.08.010.
To study the immune regulation function of high expressing interleukin-10 (IL-10) in B cells on CD4(+)T-cells in periodontitis mouse model. Twenty-four 7-weeks-old female C57BL/6 mice were randomly and equally assigned into 4 groups: the healthy control group (HC group, 6), the ligation combined (Pg) infection group (P group, 6), the ligation combined Pg infection with non-stimulated B cell transfer group (P+NSB group, 6) and the ligation combined Pg infection with stimulated B cell transfer group (P+SB group, 6). Ligation combined Pg infection of the maxillary second molar was used to induce periodontitis of mice. The exogenous non-stimulated B cells or stimulated B cells were injected into the palate gingivalat the 5th day after ligation, and all mice were sacrificed at the 14th day. HE stain was used to detect the histological of periodontal tissues, toluidine blue stain was used to analysis the alveolar bone loss, immunofluorescence stain was used to detect the expression of CD4(+)T-cell and IL-10, immunohistochemical was used to detect the expression of receptor activator of NF-κB ligand (RANKL) and IL-1β. Results of HE staining showed that more hyperplasia of gingival epithelium and the alveolar bone resorption in P group, P+NSB group and P+SB group compared with HC group. Results of toluidine blue staining showed that the alveolar bone losses in P group [(0.668±0.041) mm(2)], P+NSB group [(0.750±0.039) mm(2)] and P+SB group [(0.517±0.038) mm(2)] were significantly increased compared with that in HC group [(0.336±0.029) mm(2)](146.051, 0.01), and the alveolar bone resorption was significantly increased in P+NSB group compared with that in P group (146.051, 0.01). However, compared with P+NSB group and P group, the alveolar bone loss in P+SB groupwas significantly decreased (146.051, 0.01). Results of immunofluorescence staining showed that CD4(+)T-cells expressed in P group [(287.5±37.9) cell/mm(2)], P+NSB group [(314.6±53.3) cell/mm(2)] and P+SB group [(185.4±42.9) cell/mm(2)] were higher than that in HC group [(12.5±13.7) cell/mm(2))(71.284, 0.01). Compared with P group and P+NSB group, CD4(+)T-cells expression in group P+SB was decreased (71.284, 0.01). IL-10 levels were increased in P group [(111.7±20.4) cell/mm(2)], P+NSB group [(126.7±15.1) cell/mm(2)] and P+SB group [(191.0±22.6) cell/mm(2)] compared with that in HC group [(22.7±4.3) cell/mm(2)] (98.516, 0.01), and the IL-10 expressed in P+SB group was significantly higher than those in P+NSB group and P group. Results of immunohistochemical tests showed that RANKL expressions in gingival tissues among P group [(674.0±71.5) cell/mm(2)], P+NSB group [(831.0±97.5) cell/mm(2)] and P+SB group [(420.1±40.8) cell/mm(2)] were significantly higher than that in HC group [(69.3±29.1) cell/mm(2)] (154.886, 0.01). However, it dramatically decreased in P+SB group compared with those in P group and P+NSB group.The IL-1βexpression in P group [(447.8±40.8) cell/mm(2)], P+NSB group [(512.5±38.2) cell/mm(2)] and P+SB group [(281.6±32.4) cell/mm(2)] were significantly higher than that in HC group [(50.8±20.9) cell/mm(2)], and it also higher in P+NSB group compared with in P group. However, it decreased in P+SB group compared with those in P group and P+NSB group (221.185, 0.01). High expression IL-10 in B cell smight inhibit alveolar bone loss, RANKL and IL-1β expressions and CD4(+)T-cell infiltration through IL-10.
研究牙周炎小鼠模型中B细胞高表达白细胞介素-10（IL-10）对CD4(+)T细胞的免疫调节功能。将24只7周龄雌性C57BL/6小鼠随机等分为4组：健康对照组（HC组，6只）、结扎联合（牙龈卟啉单胞菌）感染组（P组，6只）、结扎联合牙龈卟啉单胞菌感染未刺激B细胞转移组（P+NSB组，6只）和结扎联合牙龈卟啉单胞菌感染刺激B细胞转移组（P+SB组，6只）。采用结扎联合牙龈卟啉单胞菌感染上颌第二磨牙诱导小鼠牙周炎。在结扎后第5天，将外源性未刺激B细胞或刺激B细胞注射到腭部牙龈，所有小鼠在第14天处死。采用苏木精-伊红（HE）染色检测牙周组织的组织学变化，甲苯胺蓝染色分析牙槽骨吸收情况，免疫荧光染色检测CD4(+)T细胞和IL-10的表达，免疫组织化学检测核因子κB受体活化因子配体（RANKL）和IL-1β的表达。HE染色结果显示，与HC组相比，P组、P+NSB组和P+SB组牙龈上皮增生更明显，牙槽骨吸收更严重。甲苯胺蓝染色结果显示，P组[（0.668±0.041）mm²]、P+NSB组[（0.750±0.039）mm²]和P+SB组[（0.517±0.038）mm²]的牙槽骨吸收量明显高于HC组[（0.336±0.029）mm²]（146.051，P<0.01），且P+NSB组的牙槽骨吸收量明显高于P组（146.OS1，P<0.01）。然而,与P+NSB组和P组相比，P+SB组的牙槽骨吸收量明显减少（146.051，P<0.01）。免疫荧光染色结果显示，P组[（287.5±37.9）个细胞/mm²]、P+NSB组[（314.6±53.3）个细胞/mm²]和P+SB组[（185.4±42.9）个细胞/mm²]中CD4(+)T细胞的表达高于HC组[（12.5±13.7）个细胞/mm²]（71.284，P<0.01）。与P组和P+NSB组相比，P+SB组中CD4(+)T细胞的表达降低（71.284，P<0.01）。与HC组[（22.7±4.3）个细胞/mm²]相比，P组[（111.7±20.4）个细胞/mm²]、P+NSB组[（126.7±15.1）个细胞/mm²]和P+SB组[（191.0±22.6）个细胞/mm²]中IL-10水平升高（98.516，P<0.01），且P+SB组中IL-10的表达明显高于P+NSB组和P组。免疫组织化学检测结果显示，P组[（674.0±71.5）个细胞/mm²]、P+NSB组[（831.0±97.5）个细胞/mm²]和P+SB组[（420.1±40.8）个细胞/mm²]牙龈组织中RANKL的表达明显高于HC组[（69.3±29.1）个细胞/mm²]（154.886，P<0.01）。然而，与P组和P+NSB组相比，P+SB组中RANKL的表达明显降低。P组[（447.8±40.8）个细胞/mm²]、P+NSB组[（512.5±38.2）个细胞/mm²]和P+SB组[（281.6±32.4）个细胞/mm²]中IL-1β的表达明显高于HC组[（50.8±20.9）个细胞/mm²]，且P+NSB组中IL-1β的表达高于P组。然而，与P组和P+NSB组相比，P+SB组中IL-1β的表达降低（221.185，P<0.01）。B细胞中高表达的IL-10可能通过IL-10抑制牙槽骨吸收、RANKL和IL-1β的表达以及CD4(+)T细胞浸润。
