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
{"title":"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。","authors":"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","doi":"10.1111/iej.14255","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Aim</h3>\n \n <p>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 <i>toll</i>-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.</p>\n </section>\n \n <section>\n \n <h3> Methodology</h3>\n \n <p>Cross-sectional study. Otherwise healthy individuals with AP (<i>n</i> = 25) and controls (<i>n</i> = 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).</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>TLR-2 expression was significantly upregulated in peripheral blood monocytes from individuals with AP compared to controls (<i>p</i> = 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 (<i>p</i> < 0.05). The methylated positions at −40 and −20 in TLR-2 were associated with TLR-2 transcriptional upregulation (<i>p</i> < 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.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>AP diagnosis activates peripheral blood monocytes via −40 CpG single-site methylation, independently promoting TLR-2 expression.</p>\n </section>\n </div>","PeriodicalId":13724,"journal":{"name":"International endodontic journal","volume":"58 8","pages":"1172-1183"},"PeriodicalIF":7.1000,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"TLR-2 gene methylation in peripheral blood monocytes from apical periodontitis individuals: A cross-sectional study\",\"authors\":\"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\",\"doi\":\"10.1111/iej.14255\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Aim</h3>\\n \\n <p>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 <i>toll</i>-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.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methodology</h3>\\n \\n <p>Cross-sectional study. Otherwise healthy individuals with AP (<i>n</i> = 25) and controls (<i>n</i> = 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).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>TLR-2 expression was significantly upregulated in peripheral blood monocytes from individuals with AP compared to controls (<i>p</i> = 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 (<i>p</i> < 0.05). The methylated positions at −40 and −20 in TLR-2 were associated with TLR-2 transcriptional upregulation (<i>p</i> < 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. 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TLR-2 gene methylation in peripheral blood monocytes from apical periodontitis individuals: A cross-sectional study
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.
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.
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