国际口腔医学杂志

国际口腔医学杂志 ›› 2024, Vol. 51 ›› Issue (3): 288-295.doi: 10.7518/gjkq.2024044

• 论著 • 上一篇    下一篇

血浆无细胞线粒体外线粒体DNA与牙周炎临床指标的相关性研究

杨再目1(),曹沛1,刘振华2,栾庆先1()   

  1. 1.北京大学口腔医学院·口腔医院牙周科;国家口腔医学中心 国家口腔疾病临床医学研究中心;口腔数字化医疗技术和材料国家工程实验室 口腔数字医学北京市重点实验室 北京 100081
    2.北京市中关村医院口腔科 北京 100190
  • 收稿日期:2023-08-16 修回日期:2024-02-01 出版日期:2024-05-01 发布日期:2024-05-06
  • 通讯作者: 栾庆先
  • 作者简介:杨再目，医师，博士，Email：yangzaimu@pku.org.cn

Correlation study of plasma cell-free extra-mitochondrial mitochondria DNA and periodontitis clinical parameters

Zaimu Yang1(),Pei Cao1,Zhenhua Liu2,Qingxian Luan1()   

  1. 1.Dept. of Periodontology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
    2.Dept. of Stomatology, Beijing Zhongguancun Hospital, Beijing 100190, China
  • Received:2023-08-16 Revised:2024-02-01 Online:2024-05-01 Published:2024-05-06
  • Contact: Qingxian Luan

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摘要：

目的 血浆无细胞线粒体外线粒体DNA（cf-exmtDNA）具有促炎潜能，本文探讨血浆cf-exmtDNA与牙周炎临床指标的相关性。 方法 纳入18~45岁受试者78人，其中牙周健康者11人，牙龈炎患者11人，牙周炎患者56人。检查并记录基线牙周指标、年龄、性别、体质指数（BMI）和空腹血糖（FBG）。取4 mL抗凝静脉血，采用二次离心法提取其中cf-exmtDNA，使用实时荧光定量聚合酶链式反应检测cf-exmtDNA拷贝数。比较不同牙周炎症状态组血浆cf-exmtDNA水平，并对血浆cf-exmtDNA与平均探诊深度（mPD）、平均附着水平（mCAL）、平均出血指数（mBI）、平均菌斑指数（mPLI）、年龄、FBG、BMI等指标进行相关性分析以及多重线性回归分析。 结果 牙周炎组血浆cf-exmtDNA水平显著高于牙周健康组（P=0.042）；样本整体血浆cf-exmtDNA水平与年龄（P=0.023）、mPD（P<0.001）、mCAL（P=0.006）、mBI（P=0.026）呈正相关关系；多重回归分析中，血浆cf-exmtDNA水平主要取决于mPD。 结论 在18~45岁人群中，牙周炎患者血浆cf-exmtDNA水平较牙周健康者显著升高，血浆cf-exmtDNA水平与年龄、mPD、mCAL、mBI呈正相关关系。

关键词: 牙周炎, 无细胞DNA, 线粒体DNA, 横断面研究, 牙周临床指标

Abstract:

Objective Plasma cell-free extra-cellular mitochondrial DNA (cf-exmtDNA) shows pro-inflammatory potential. This study aims to investigate the correlation between cf-exmtDNA and baseline periodontitis clinical parameters in patients in general health. Methods A total of 78 participants aged 18-45 years were enrolled: 11 periodontal healthy volunteers, 11 patients with gingivitis, and 56 patients with periodontitis. Baseline periodontal index, age, gender, body mass index (BMI), and baseline fasting blood glucose (FBG) were examined and recorded. Anticoagulated venous blood (4 mL) was collected, and cf-mtDNA was extracted by secondary centrifugation. cf-exmtDNA concentration was measured through real-time quantitative polymerase chain reaction. Different states of periodontal inflammation were compared in terms of plasma cf-exmtDNA copy number, and the relationship among plasma cf-exmtDNA, mean probing depth (mPD), mean clinical attachment level (mCAL), mean bleeding index (mBI), mean plaque index, age, FBG, and BMI were examined through correlation and multiple linear regression analyses. Results Patients with periodontitis had significantly higher plasma cf-exmtDNA levels than the healthy volunteers (P=0.042). The overall plasma cf-exmtDNA was significantly positively correlated with age (P=0.023), mPD (P<0.001), mCAL (P=0.006), and mBI (P=0.026). Multiple regression analysis showed that plasma cf-exmtDNA level was significantly dependent on mPD. Conclusion In the general population aged 18–45, patients with periodontitis had significantly higher plasma cf-exmt-DNA level than the healthy volunteers, and plasma cf-exmtDNA concentration was significantly positively correlated with age, mPD, mCAL, and mBI.

Key words: periodontitis, cell-free DNA, mitochondrial DNA, cross-sectional study, periodontal clinical parameters

中图分类号: 

  • R781.4

表 1

qPCR的程序设定"

程序温度/℃时长
保持期502 min
预变性9510 min
PCR期（40 个循环）变性9510 s
退火6015 s
延伸a7230 s
熔解曲线期

表 2

引物序列"

基因方向序列
ND1正向5’-CACACTAGCAGAGACCAACCGAAC-3’
反向5’-CGGCTATGAAGAATAGGGCGAAGG-3’

表 3

牙周健康、牙龈炎、牙周炎组基线特征"

纳入指标组别
牙周健康组（n=11）牙周炎组（n=56）
男/女4/78/330/26
年龄/岁25（23，29）26（24，30）32（26，35）
空腹血糖/（mmol/L）4.80±0.295.05±0.064.96±0.17
体质指数/（kg/m2）20.89±4.7422.22±6.8221.27±4.73
mPD/mm2.16（2.11，2.39）2.35（2.10，2.40）2.97（2.73，3.67）
mCAL/mm0.03（0.00，0.03）0.03（0.00，0.05）3.07（2.77，3.80）
mBI1.32±0.191.66±0.292.53±0.34
mPLI1.15±0.241.40±0.621.55±0.25

图1

血浆cf-exmtDNA与牙周炎症状态和临床指标的关系A：血浆cf-exmtDNA拷贝数；B、C、D、E分别为血浆cf-exmtDNA与mPD、mCAL、mBI、年龄的Pearson相关性分析；*：P<0.05；ns：P>0.05。"

表 4

各临床指标与血浆cf-exmtDNA的相关性"

测量指标与血浆cf-exmtDNA的相关系数
r值P值
牙周指标mPD/mm0.409<0.001
mCAL/mm0.3100.006
mBI0.2520.026
mPLI0.0090.936
全身指标年龄/岁0.2570.023
BMI /（kg/m2）0.2060.071
FBG/（mmol/L）-0.0290.802

表 5

血浆cf-exmtDNA和牙周临床指标的多重线性回归分析"

多重线性回归模型模型参数B95% CIβt值P值
血浆cf-exmtDNA常量-1 257.091（-2 090.559，-423.622）-3.0040.004
mPD528.636（258.892，798.380）0.4093.903<0.001
R2adj=0.156（n=78，P<0.001）
mPD常量1.117（0.774，1.459）6.491<0.001
血浆cf-exmtDNA1.73×10-4（0.68×10-4，2.77×10-4）0.2233.2850.002
mBI0.811（0.662，0.960）0.73710.841<0.001
R2adj=0.667（n=78，P<0.001）
mBI常量-0.223（-0.626，0.180）-1.1030.273
mPD0.672（0.551，0.793）0.73911.056<0.001
mPLI0.310（0.136，0.484）0.2373.5530.001
R2adj=0.674（n=78，P<0.001）

图2

多变量气泡图（左）与mPD的多重线性回归模型（右）"

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