口腔医学研究

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (9): 962-965.DOI: 10.13701/j.cnki.kqyxyj.2017.09.013

• 基础研究论著 • 上一篇    下一篇

具核梭杆菌对牙龈卟啉单胞菌或伴放线聚集杆菌诱导多形核白细胞活性氧生成的影响

李，杨春瑜，刘宗霞，张彦表，梁广智*   

  1. 潍坊医学院口腔内科学教研室 山东 潍坊 261053
  • 收稿日期:2017-04-10 出版日期:2017-09-20 发布日期:2017-09-27
  • 通讯作者: 梁广智,E-mail: kqnk@wfmc.edu.cn
  • 作者简介:李(1978~ ),女,山东淄博人,博士,讲师,主要从事临床牙周病学的工作。

Effect of Co-infection with Fusobacteriumnucleatum on Production of Reactive Oxygen Species from Polymorphonuclear Cells Stimulated by Porphyromonas Gingivalis or Aggregatibacter Actinomycetemcomitans

LI Yan, YANG Chun-yu, LIU Zong-xia, ZHANG Yan-biao, LIANG Guang-zhi   

  1. Department of Oral Medicine, Weifang Medical University, Weifang 261053, China.
  • Received:2017-04-10 Online:2017-09-20 Published:2017-09-27

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摘要： 目的:探讨与具核梭杆菌(F. nucleatum,Fn)共同感染时,牙龈卟啉单胞菌(P. gingivalis,Pg)或伴放线聚集杆菌(A.actinomycetemcomitans,Aa)对多形核白细胞(PMNs)产生活性氧(ROS)的影响。方法:取健康人抗凝外周血,经Percoll分离后取PMNs,将分离的PMNs与2’,7’-二氯荧光素二乙酸酯(DCFH-DA)混合,于37 ℃的水浴箱中孵育15 min,然后分别向管中加入A组和B组细菌(细菌分组:A1, Pg;A2, Aa;A3,Fn;B1, Pg+Fn;B2, Aa+Fn;B3, Pg+Aa;B4, Pg+Aa+Fn;B5, Aa+Pg+Fn),充分混合后于37 ℃孵育1 h和4 h,流式细胞术 (Flow Cytometry, FCM)检测PMNs产生ROS的量。结果:不同牙周致病菌与分离的PMNs在37 ℃作用1 h,PMNs生成ROS的量分别是A1-16.52;A2-18.74;A3-19.59;B1-17.60;B2-20.09,其中B1>A1;B2>A2,差异显著(P<0.05)。不同微生物与分离的PMNs在37 ℃作用4 h,PMNs生成ROS的量分别是A1-176.05;A2-203.19;A3-230.36;B1-259.08;B2-274.89,其中B1>A1;B2>A2,差异显著(P<0.05)。结论:与Fn共聚后,Pg或Aa诱导PMNs产生ROS的能力被增强。

关键词: 多形核白细胞, 活性氧, 具核梭杆菌, 牙龈卟啉单胞菌, 伴放线聚集杆菌

Abstract: Objective: To investigate the production of reactive oxygen species (ROS) from polymorphonuclear cells (PMNs) stimulated by porphyromonas gingivalis (P. gingivalis, Pg) or aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans, Aa) when co-infected with fusobacterium nucleatum (F. nucleatum, Fn) . Methods: Venous blood of healthy volunteers was obtained with heparin anticoagulation, the PMNs isolated with Percoll was mixed separated with 2’,7’-Dichlorodihydrofluorescindiacetate (DCFH-DA) and then incubated for 15 min in water bath at 37℃. Bacteria of group A and B (groups: A1, Pg; A2, Aa; A3, Fn;B1, Pg + Fn; B2, Aa + Fn; B3, Pg + Aa; B4, Pg + Aa + Fn; B5, Aa + Pg + Fn)were added into the tube for sufficient mixing, and then was incubated (37℃) for 1h and 4h. The amount of ROS produced by PMNs was measured by flow cytometry. Results: After the reaction of periodontopathic bacteria and isolated PMNs at 37℃ for 1h and 4h, the ROS production amounts were respectively A1-16.52; A2-18.74; A3-19.59; B1-17.60; B2-20.09. Among them, B1>A1, and B2>A2 (P<0.05). At the time of 4h, the amounts of ROS production were respectively A1-176.05; A2-203.19; A3-230.36; B1-259.08; B2-274.89. Among them, B1>A1 and B2>A2 (P<0.05). Conclusion: These results suggested that the production of ROS from PMNs by Pg and Aa can be enhanced when co-infected with Fn.

Key words: Polymorphonuclear cells , Reactive oxygen species, Fusobacteriumnucleatum, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans

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