TLR-2 gene methylation in peripheral blood monocytes from apical periodontitis individuals: A cross-sectional study

IF 7.1 1区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

International endodontic journal Pub Date : 2025-05-23 DOI:10.1111/iej.14255

María José Bordagaray, Alejandra Fernández, Elizabeth Pellegrini, Mauricio Garrido, Patricia Hernández, David González-Quintanilla, Gabriel Navia, Daniela Rehbein, Mauricio Baeza, Peter Gebicke-Haerter, Marcela Hernández

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引用次数: 0

Abstract

Aim

Apical periodontitis (AP) is a chronic inflammatory disease arising from the contamination of the root canal system. Epigenetic regulation plays a pivotal role in controlling monocyte/macrophage-mediated local and systemic responses to bacterial challenges via toll-like receptors (TLRs). We aimed to explore the relationship between the methylation and expression patterns of TLR-2 in peripheral blood monocytes of individuals with AP and controls.

Methodology

Cross-sectional study. Otherwise healthy individuals with AP (n = 25) and controls (n = 29) were recruited. Peripheral blood monocytes were isolated from blood samples by Ficoll gradient and negative immunoselection. DNA and RNA were extracted from monocytes. DNA was bisulfite-treated, amplified and sequenced to evaluate the global and cytosine-phosphate-guanine (CpG) single-site methylation pattern of the TLR-2 gene promoter. mRNA relative levels of TLR-2 were assessed by qPCR. The potential associations between AP, TLR-2 DNA methylation and TLR-2 gene expression were explored using generalized structural equation models (GSEM).

Results

TLR-2 expression was significantly upregulated in peripheral blood monocytes from individuals with AP compared to controls (p = 0.005). Though no differences were found in the global methylation pattern, CpG single sites from the TLR-2 gene were differentially methylated at positions −40 and +24 (p < 0.05). The methylated positions at −40 and −20 in TLR-2 were associated with TLR-2 transcriptional upregulation (p < 0.05). Further evaluation with GSEM analysis showed that AP promoted the methylation of the −40 CpG single site on the TLR-2 gene, which, in turn, upregulated TLR-2. Conversely, the methylation of the −20 CpG single site did not act as a mediator of TLR-2 transcription in AP.

Conclusions

AP diagnosis activates peripheral blood monocytes via −40 CpG single-site methylation, independently promoting TLR-2 expression.

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根尖牙周炎患者外周血单核细胞TLR-2基因甲基化：一项横断面研究。

目的：根尖牙周炎（AP）是由根管系统污染引起的慢性炎症性疾病。表观遗传调控在通过toll样受体（TLRs）控制单核细胞/巨噬细胞介导的局部和全身对细菌攻击的反应中起关键作用。我们旨在探讨AP患者和对照组外周血单核细胞中TLR-2的甲基化和表达模式之间的关系。方法：横断面研究。此外，还招募了健康的AP患者（n = 25）和对照组（n = 29）。采用Ficoll梯度法和阴性免疫选择法分离外周血单核细胞。从单核细胞中提取DNA和RNA。对DNA进行亚硫酸处理、扩增和测序，以评估TLR-2基因启动子的全局和胞嘧啶-磷酸-鸟嘌呤（CpG）单位点甲基化模式。采用qPCR检测TLR-2 mRNA相对水平。利用广义结构方程模型（GSEM）探讨了AP、TLR-2 DNA甲基化和TLR-2基因表达之间的潜在关联。结果：与对照组相比，AP患者外周血单核细胞中TLR-2的表达显著上调（p = 0.005）。尽管在整体甲基化模式上没有发现差异，但来自TLR-2基因的CpG单位点在-40和+24位点的甲基化存在差异(p)结论：AP诊断通过-40 CpG单位点甲基化激活外周血单核细胞，独立促进TLR-2的表达。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

International endodontic journal 医学-牙科与口腔外科

CiteScore

10.20

自引率

28.00%

发文量

195

审稿时长

4-8 weeks

期刊介绍： The International Endodontic Journal is published monthly and strives to publish original articles of the highest quality to disseminate scientific and clinical knowledge; all manuscripts are subjected to peer review. Original scientific articles are published in the areas of biomedical science, applied materials science, bioengineering, epidemiology and social science relevant to endodontic disease and its management, and to the restoration of root-treated teeth. In addition, review articles, reports of clinical cases, book reviews, summaries and abstracts of scientific meetings and news items are accepted. The International Endodontic Journal is essential reading for general dental practitioners, specialist endodontists, research, scientists and dental teachers.