International endodontic journal · 2025
TLR-2 gene methylation in peripheral blood monocytes from apical periodontitis individuals: A cross-sectional study.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
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Bordagaray María José, Fernández Alejandra, Pellegrini Elizabeth, Garrido Mauricio, Hernández Patricia, González-Quintanilla David, Navia Gabriel, Rehbein Daniela, Baeza Mauricio, Gebicke-Haerter Peter, Hernández Marcela
Аннотация
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.
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.