慢性牙周炎患者唾液诱导巨噬细胞分化和活化的研究
Saliva of periodontitis patients promotes macrophage differentiation and activation
作者：李小英,李倩,崔博淼,王利伟,樊亚平,陈娇,冯云,张平
Author：Xiaoying Li,Qian Li,Bomiao Cui,Liwei Wang,Yaping Fan,Jiao Chen,Yun Feng,Ping Zhang
收稿日期：2018-03-08 年卷（期）页码：2018,36(5):508-508-513
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：唾液,慢性牙周炎,巨噬细胞,
Key words：saliva,chronic periodontitis,macrophage,
基金项目：国家自然科学基金(81372892);口腔疾病研究国家重点实验室自主研究课题(SKLOD201601)
中文摘要
目的 探讨慢性牙周炎（CPD）唾液对巨噬细胞分化、活化、吞噬功能和分泌破骨细胞成熟因子的影响。方法 收集20名正常人和20名CPD患者的唾液样品。15%唾液刺激外周血单个核细胞（PBMCs）和人单核细胞系（THP-1）5 d,用流式细胞仪检测淋巴细胞亚群变化,显微镜观察巨噬细胞分化和吞噬活性;酶联免疫吸附测定（ELISA）检测巨噬细胞分泌破骨细胞成熟因子情况。结果 CPD唾液刺激PBMCs 5 d后,61.25%±11.33%的PBMCs分化为大颗粒细胞,其中86.78%±13.69%的大颗粒细胞为CD14 ++CD16 +巨噬细胞;CPD唾液可诱导THP-1细胞分化为巨噬细胞,表达高水平CD14,分泌高水平破骨细胞成熟因子;CPD唾液诱导后THP-1细胞的吞噬作用（66.35%±9.67%）明显高于正常唾液组（40.71%±3.52%）和阴性对照组（33.33%±7.52%）。结论 CPD唾液能够诱导巨噬细胞的分化和活化,促进巨噬细胞吞噬细菌,分泌破骨细胞成熟因子。
英文摘要
ObjectiveThe aim of this study was to investigate the effect of saliva of patients with chronic periodontitis (CPD) on the differentiation, activation, and secretion of osteoclast-maturing mediators of macrophages. Methods A total of 40 saliva samples were collected from healthy donors (n=20) and severe periodontitis patients (n=20). Peripheral blood mono-nuclear cells (PBMCs) and THP-1 monocyte line cells were challenged with 15% saliva for 5 days. The phenotype, surface marker, and phagocytosis of macrophages were analyzed by flow cytometry and microscopy. Osteoclast-maturing mediators were assayed by using enzyme-linked immunosorbent assay (ELISA) kits. Results When PBMCs were treated with CPD saliva for 5 days, 61.25%±11.33% of cells were transformed into large granular cells; 86.78%±13.69% of large granular cells were identified as CD14++CD16+macrophages. When THP-1 cells were treated with CPD saliva, most cells attached to thebottom of cell culture plates, thereby exhibiting macrophage morphology and releasing additional osteoclast-maturing mediators. Furthermore, the phagocytosis of THP-1 cells considerably increased in the presence of CPD saliva (66.35%±9.67%) compared with medium control (33.33%±7.52%), or healthy saliva (40.71%±3.52%).Conclusion Saliva from patients with CPD can induce macrophage differentiation, activate phagocytose microorganisms, and secrete osteoclast-maturing mediators.
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