当前位置: X-MOL 学术Int. Endod. J › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Epigenetic regulation of TLR2-mediated periapical inflammation.
International Endodontic Journal ( IF 5 ) Pub Date : 2020-06-19 , DOI: 10.1111/iej.13329
A Fernández 1, 2 , P Veloso 1 , J Astorga 1 , C Rodríguez 2 , V A Torres 3 , M Valdés 4 , M Garrido 1 , P J Gebicke-Haerter 5, 6 , M Hernández 1, 7
Affiliation  

AIM To determine the methylation pattern of TLR2 gene promoter and its association with the transcriptional regulation of periapical inflammatory and angiogenic responses in symptomatic and asymptomatic forms of apical periodontitis. METHODOLOGY In this cross-sectional study, apical lesions were obtained from volunteers with asymptomatic apical periodontitis (AAP) (n = 17) and symptomatic apical periodontitis (SAP) (n = 17) scheduled for tooth extraction, and both total RNA and DNA were extracted. DNA was bisulfite-treated, a region of CpG island within the TLR2 gene was amplified by qPCR and the products were sequenced. Additionally, the mRNA expression of TLR2, TLR4, IL-6, IL-12, TNFalpha, IL-23, IL-10, TGFbeta, VEGFA and CDH5 was analysed by qPCR. The data were analysed with chi-square tests, Mann-Whitney or unpaired t-tests, and Spearman´s correlation; variable adjustments were performed using multiple linear regression (P < 0.05). RESULTS TLR2 depicted a hypomethylated DNA profile at the CpG island in SAP when compared with AAP, along with upregulated expression of TLR2, with pro-inflammatory cytokines IL-6 and IL-23, and the angiogenesis marker CDH5 (P < 0.05). TLR2 methylation percentage negatively correlated with mRNA levels of IL-23 and CDH5 in apical periodontitis. Lower methylation frequencies of single CpG dinucleotides -8 and -10 localized in close proximity to nuclear factor κB (NFκB) binding within the TLR2 promoter were identified in SAP versus AAP (P < 0.05). Finally, unmethylated -10 and -8 single sites demonstrated up-regulation of IL-23, IL-10 and CDH5 transcripts compared to their methylated counterparts (P < 0.05). CONCLUSIONS TLR2 gene promoter hypomethylation was linked to transcriptional activity of pro-inflammatory cytokines and angiogenic markers in exacerbated periapical inflammation. Moreover, unmethylated single sites in close proximity to NFκB binding were involved in active transcription of IL-23, IL-10 and CDH5.

中文翻译:

TLR2介导的根尖周炎症的表观遗传调控。

目的 确定 TLR2 基因启动子的甲基化模式及其与有症状和无症状形式的根尖周炎中根尖周炎症和血管生成反应的转录调控的关系。方法 在这项横断面研究中,根尖病变来自于计划拔牙的无症状根尖周炎 (AAP) (n = 17) 和有症状根尖周炎 (SAP) (n = 17) 的志愿者,总 RNA 和 DNA提取。DNA经过亚硫酸氢盐处理,通过qPCR扩增TLR2基因内的CpG岛区域并对产物进行测序。此外,通过 qPCR 分析 TLR2、TLR4、IL-6、IL-12、TNFα、IL-23、IL-10、TGFβ、VEGFA 和 CDH5 的 mRNA 表达。使用卡方检验、Mann-Whitney 或非配对 t 检验分析数据,和斯皮尔曼的相关性;使用多元线性回归进行变量调整(P <0.05)。结果 与 AAP 相比,TLR2 描绘了 SAP 中 CpG 岛的低甲基化 DNA 谱,同时 TLR2 表达上调,具有促炎细胞因子 IL-6 和 IL-23,以及血管生成标志物 CDH5(P < 0.05)。TLR2甲基化百分比与根尖周炎中IL-23和CDH5的mRNA水平呈负相关。在 SAP 与 AAP 中,确定了位于 TLR2 启动子内的核因子 κB (NFκB) 结合附近的单个 CpG 二核苷酸 -8 和 -10 的较低甲基化频率(P < 0.05)。最后,与甲基化对应物相比,未甲基化的 -10 和 -8 单位点显示出 IL-23、IL-10 和 CDH5 转录物的上调(P < 0.05)。结论 TLR2 基因启动子低甲基化与加剧根尖周炎症的促炎细胞因子和血管生成标志物的转录活性有关。此外,靠近 NFκB 结合的未甲基化单个位点参与了 IL-23、IL-10 和 CDH5 的主动转录。
更新日期:2020-05-19
down
wechat
bug