B淋巴细胞对牙周致病菌免疫反应及骨细收因子RANKL表达的影响  

The effect of B lymphocytes on immune response of periodontal bacteria and osteoclast differentiated reactor-RANKL expression

作　　者：林晓萍[1,2] 韩晓哲 魏巍[1] Martin A. Taubman 

机构地区：[1]中国医科大学附属盛京医院口腔科 [2]Department of Immunology,The Forsyth Institute,Boston,MA 02115,USA

出　　处：《上海口腔医学》2009年第4期392-396,共5页Shanghai Journal of Stomatology

基　　金：辽宁省自然科学基金(20042062);辽宁省教育厅科研基金(202013124)~~

摘　　要：目的:通过评价B淋巴细胞与牙周致病菌——伴放线放线杆菌(Aa)发生免疫反应过程中核因子-КB受体活化配体(RANKL)的表达,探讨B淋巴细胞是否参与Aa引起的牙周骨吸收。方法:采用RT-PCR方法测定大鼠脾细胞中第1和7d细胞因子的mRNA转录水平,采用流式细胞技术测定大鼠脾细胞中B细胞表达RANKL IgG阳性细胞的百分数,应用TRAP法评价B淋巴细胞对破骨细胞分化的诱导潜力。实验结果采用SPSS 10.0软件包进行分析。结果:在无Aa抗原刺激的条件下,大鼠脾细胞培养1d和7d的TNF-α及IL-4表达水平均升高,IL-10、RANKL的表达水平无明显变化;加入Aa组培养1d时,TNF-α的表达显著增加,7d后,IL-4、IL-10及RANKL的mRNA转录水平显著增加;B细胞的百分数以及表达RANKL的IgG阳性细胞百分数与不加Aa组相比显著增加;Aa免疫组加入Aa培养后,表达RANKL的IgG阳性细胞百分数增加显著高于非免疫组。Aa免疫组的B淋巴细胞与RAW 264.7细胞共同培养后,TRAP染色阳性细胞显著增加(P<0.01);加入人OPG-Fc培养,可显著抑制TRAP阳性细胞的形成(P<0.05)。结论:B淋巴细胞经特异性抗原Aa活化后,通过上调RANKL的表达,增加对破骨细胞分化的诱导潜力,参与牙周骨吸收。PURPOSE：To study whether B lymphocytes involve in bone resorption in periodontal disease by evaluating RANKL expression of B lymphocytes in immune response to aggregatibacter actinomycetemcomitans（Aa）.METHODS：The expression of mRNA transcripts involved in day 1 and day 7 responses of cytokines in cultured rat splenocytes was determined by RT-PCR;The percentage of RANKL-expressing IgG positive cells by B cells in cultured rat splenocytes was measured by flow cytometry;and B lymphocytes capacity for induction of osteoclast differentiation was evaluated by TRAP staining.SPSS 10.0 software package was used for statistical analysis.RESULTS：The expression of IL-4 and IL-10 in cultured cells was not changed in the presence of Aa on day 1,after culture for 7 days;The percentage of B lymphocytes,all RANKL mRNA transcripts and percentage of RANKL-expressing IgG positive cells were significantly increased in the presence,compared to the absence of Aa.These increases were considerably greater in cells isolated from immunized than non-immunized rats.B cells from Aa immunized rats significantly increased induction of osteoclast differentiation（P〈0.01）,and addition of human OPG-Fc into the culture significantly inhibited its induction（P〈0.05）,and was RANKL-independent.CONCLUSION：B lymphocytes contribute to bone resorption in periodontal disease throughinvolving in the immune response to Aa,up-regulation of RANKL expression and increasing induction of osteoclast differentiation.

关 键 词：牙周病 动物模型 核因子-КB受体活化配体 B淋巴细胞 免疫反应 伴放线放线杆菌 

分 类 号：R781.4[医药卫生—口腔医学]