国际口腔医学杂志

国际口腔医学杂志 ›› 2019, Vol. 46 ›› Issue (5): 532-539.doi: 10.7518/gjkq.2019052

• 论著 • 上一篇    下一篇

16S核糖体DNA高通量测序研究种植体龈沟液微生物的变化

青薇1,黄丽娟1,郑佳俊1,任静1,李成龙2,庹嫱2,任小华3,牟雁东1,3()   

  1. 1. 西南医科大学口腔医学院 泸州 646099
    2. 电子科技大学医学院 成都 610054
    3. 四川省人民医院口腔科 成都 610072
  • 收稿日期:2018-09-05 修回日期:2019-04-25 出版日期:2019-09-01 发布日期:2019-09-10
  • 通讯作者: 牟雁东
  • 作者简介:青薇,医师,硕士,Email： 513866210@qq.com
  • 基金资助:
    四川省科学技术厅项目(2016TD0008);中央高校基金(A03017023801295)

Shift of microbial composition of peri-implant gingival crevicular fluid as revealed by 16S ribosomal DNA high-throughput sequencing

Qing Wei1,Huang Lijuan1,Zheng Jiajun1,Ren Jing1,Li Chenglong2,Tuo Qiang2,Ren Xiaohua3,Mu Yandong1,3()   

  1. 1. School of Stomatology, Southwest Medical University, Luzhou 646099, China
    2. School of Medicine, University of Electronic Science and Technology, Chengdu 610054, China
    3. Dept. of Stomatology, Sichuan Provincial People’s Hospital, Chengdu 610072, China
  • Received:2018-09-05 Revised:2019-04-25 Online:2019-09-01 Published:2019-09-10
  • Contact: Yandong Mu
  • Supported by:
    This study was supported by Sichuan Science and Technology Department Project(2016TD0008);Central University Fund(A03017023801295)

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摘要：

目的 应用16S核糖体DNA（rDNA）高通量测序技术研究健康天然牙、健康种植体以及种植体周围炎的植体龈沟液内微生物菌群的差异,为理解种植体周围组织从健康至疾病状态的病因提供依据。方法 通过Illumina HiSeq 2500测序技术,分析30个龈沟液细菌的菌群结构及多样性。结果 种植体周围炎组Shannon指数低于健康牙及健康种植体组（P<0.05）,细菌群落主要属于厚壁菌门（Firmicutes）、变形菌门（Proteobacteria）、梭杆菌门（Fusobacteria）、拟杆菌门（Bacteroidetes）、放线菌门（Actinobacteria）等17个门,普氏菌属（Prevotella）、密螺旋体属（Treponema）、纤毛菌属（Leptotrichia）、放线菌属（Actinomyces）、链球菌属（Streptococcus）和丁酸弧菌属（Butyrivibrio）等497个属。优势菌门以拟杆菌门在种植体周围炎组相对较高（P<0.05）,属水平上丁酸弧菌为种植体周围炎的优势菌属之一,代谢产物异丁酸的增多可能是导致疾病状态的一个因素。结论 健康种植体与健康天然牙一致,呈现出丰富的微生物多态性,而当周围炎形成后,表现为相关致病菌丰度增高,而其多样性下降。

关键词: 高通量测序, 种植体周围炎, 微生物多样性

Abstract:

Objective Microbes in gingival crevicular fluid are the key components of the subgingival microecosystem around implants and are closely related to implant health. Methods The microbial structure and diversity of 30 gingival crevicular bacteria were analysed by Illumina HiSeq 2500 sequencing technology. Results The Shannon indices of the peri-implantitis group were lower than those of healthy natural teeth and healthy implants (P<0.05). The bacterial communities mainly belong to Firmicutes, Proteobacteria, Fusobacteria, Bacteroidetes, Actinobacteria and 12 other phyla, and Prevotella, Treponema, Leptotrichia, Actinomyces, Streptococcus, Butyrivibrio and 491 other genera. At the phylum level, the Bacteroides was relatively high in the periimplan-titis group (P<0.05). Butyrivibrio is one of the dominant bacteria in peri-implant inflammation. The increase in isobutyric acid may be one of the factors leading to disease state. Conclusion Healthy implants exhibit abundant microbial polymorphisms, which are consistent with healthy natural teeth. When the tissues around the implants are exposed to inflammation caused by pathogenic bacteria, the abundance of related pathogens is increased, and its diversity declines.

Key words: high-throughput sequencing, peri-implantitis, microbial diversity

中图分类号: 

  • Q257

表 1

患者基本特征及临床检查指标"

项目 健康天然牙 健康种植体 种植体周围炎
牙位 上颌前牙 2 2 2
上颌后牙 4 4 5
下颌前牙 1 1 0
下颌后牙 3 3 3
行使功能时间/d 5.7±0.7 4.2±3.8
PD/mm 2.4±1.9 2.9±1.7 6.0±3.5
BOP 1 2 10

图 1

原始数据长度分布图"

图 2

样本稀释曲线图2.3 各样品中细菌多样性及相关性分析"

表 2

3组样本的Alpha多样性指数"

组别 Chao 1 ACE Simpson Shannon Coverage
H 8 198.80±284.10 12 020.69±531.10 0.06±0.02 5.40±0.19 0.91±0.01
HI 8 985.73±1 494.03 13 496.45±497.50 0.06±0.02 5.47±0.21 0.91±0.01
PI 8 181.59±428.92 13 189.68±497.50 0.07±0.02 5.24±0.13 0.92±0.01
H-HI P值 0.281 0.161 0.672 0.466 0.622
H-PI P值 0.281 0.139 0.457 0.041* 0.645
HI-PI P值 0.942 0.795 0.245 0.009* 0.424

图 3

3组的Shannon指数差异2.4 主成分分析"

图 4

主成分分析2.5 门水平菌群组成分析"

图 5

样品在门分类水平上细菌类群比较 注：各组例数n=10。*与PI组相比,P<0.05。"

图 6

优势菌门的比较*与PI组相比,P<0.05。"

图 7

属水平的热图比较"

图 8

样品在属分类水平上细菌类群比较*PI组与HI组对比,P<0.01;#PI组与H组对比,P<0.01。"

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