牙龈卟啉单胞菌脂多糖对树突状细胞成熟及功能影响的体外研究  

Effects of Porphyromonas gingivalis-lipopoly saccharide on the maturation and functions of dendritic cells

作　　者：苏寒[1] 毛钊[1] 陈伟[1] 郭婷[1] 闫翔[2] 

机构地区：[1]南京军区南京总医院口腔科,南京210002 [2]南京大学医学院附属口腔医院正畸科,南京210008

出　　处：《东南国防医药》2017年第5期465-472,共8页Military Medical Journal of Southeast China

基　　金：南京市医学科技发展重点项目(zkx15035);南京市第七批科技发展计划项目(201507043)

摘　　要：目的研究牙龈卟啉单胞菌脂多糖(P.gingivalis-LPS)的刺激对大鼠树突状细胞(DCs)成熟及功能的影响,为探索DCs在牙周炎的发生发展中的作用机制提供实验依据。方法采用流式细胞学方法检测P.gingivalis-LPS和大肠杆菌脂多糖(E.coli-LPS)刺激下,CD11c^+MHCⅡ^+、CD11c^+CD80^+、CD11c^+CD86^+和CD11c^+CD40^+DCs的比率;采用ELISA法检测DCs分泌白介素-12(IL-12)、干扰素-γ(IFN-γ)、白介素-10(IL-10)和白介素-13(IL-13)的量。采用CCK8法检测与上述DCs共培养的CD4+T细胞的增殖;采用ELISA法检测T细胞分泌IL-2、IFN-γ、IL-10和IL-13的量。在上述的培养系统中加入Toll样受体4(TLR4)抑制剂(polymyxin B,PmB)或TLR2/TLR4抑制剂(oxidation of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine,OxPAPC),观察TLR抑制剂对上述DCs成熟及功能的影响。结果 P.gingivalis-LPS与E.coli-LPS均能刺激DCs成熟。TLR4抑制剂明显抑制E.coli-LPS组DCs成熟和抗原提呈功能,对P.gingivalis-LPS组DCs成熟和抗原提呈功能没有显著抑制。TLR2/TLR4抑制剂对P.gingivalis-LPS组DCs成熟和抗原提呈功能显著抑制。P.gingivalis-LPS组DCs分泌IL-12和IFN-γ的量低于E.coli-LPS组(P<0.05);P.gingivalis-LPS组DCs分泌IL-10和IL-13的量高于E.coli-LPS组(P<0.05)。与P.gingivalis-LPS和E.coli-LPS共培养的DCs均能促进CD4+T细胞增殖。与P.gingivalis-LPS组DCs共培养的T细胞分泌IL-2和IFN-γ的量低于E.coli-LPS组(P<0.05);其分泌IL-10的量高于E.coli-LPS组(P<0.05)。结论 P.gingivalis-LPS能促进DCs的成熟和抗原提呈功能。P.gingivalis-LPS刺激下的DCs促进Th2型免疫应答;E.coli-LPS刺激下的DCs促进Th1型免疫应答。P.gingivalis-LPS通过TLR2通路刺激DCs成熟;E.coli-LPS通过TLR4通路刺激DCs成熟。Objective dy the roles of P.gingivalis-LPS on the maturation and functions of DCs to provide experimental evidences to explore the possible mechanism of DCs in periodontitis. Methods Flow cytometry was used to detect CD11c,MHCⅡ,CD80,CD86 and CD40 expression on DCs which were stimulated by P.gingivalis-LPS or E. coli-LPS and ELISA was used to detect IL-12,IFN-γ,IL-10 and IL-13 secreted by DCs. CCK8 was used to assay CD4-＋T cells proliferation after co-cultured with DCs stimulated by P.gingivalis-LPS or E.coli-LPS and ELISA was used to detect IL-2,IFN-γ,IL-10 and IL-13 secreted by T cells. TLR4 inhibitor（ polymyxin B） or TLR2 and TLR4 inhibitor（ OxPAPC） was added to P.gingivalis-LPS group and E.coli-LPS group to observe the effects of these two TLR inhibitors on the maturation and antigenpresenting functions of DCs. Results The capacity of P.gingivalisLPS to stimulate DCs maturation was similar to that of E.coliLPS. When TLR4 inhibitor was added to E. coli-LPS group,maturation and antigen-presenting functions of DCs were significantly inhibited. When TLR4 inhibitor was added to P. gingivalis-LPS group,maturation and antigen-presenting functions of DCs were not significantly inhibited. When TLR2 and TLR4 inhibitor was added to P.gingivalis-LPS group,maturation and antigen-presenting functions of DCs were significantly inhibited.The level of IL-12 and IFN-γ secreted by DCs in P. gingivalis-LPS group was significantly lower than that of E. coli-LPS group（ P〈0.05）,meanwhile,IL-10 and IL-13 secreted by DCs in P.gingivalis-LPS group was significantly higher than that of E.coliLPS group（ P〈0.05）. DCs stimulated by both P.gingivalis-LPS and E.coli-LPS could promote the proliferation of CD4-＋T cells. The amount of IL-2 and IFN-γ secreted by T cells stimulated by DCs in P.gingivalis-LPS group was significantly lower than that of E.coliLPS group（ P〈0.05）,meanwhile,IL-10 secreted by T cells stimulated by DCs in P.gingivalis-LPS group was significantly higher than that of E.coli-LPS group（

关 键 词：树突状细胞 牙龈卟啉单胞菌脂多糖 大肠杆菌脂多糖 细胞表型 抗原提呈 

分 类 号：R780.2[医药卫生—口腔医学]