Department of Morphology Division of Histology, Piracicaba Dental School, State University of Campinas, Av. Limeira 901, Caixa Postal 052, CEP 13414-903, Piracicaba, São Paulo, Brazil.
Clin Oral Investig. 2013 May;17(4):1279-85. doi: 10.1007/s00784-012-0816-z. Epub 2012 Aug 9.
The role of epigenetic regulation in inflammatory diseases such as periodontitis is poorly known. The aim of this study was to assess whether Porphyromonas gingivalis lipopolysaccharide (LPS) can modulate gene expression levels of the some enzymes that promote epigenetic events in cultures of the human keratinocytes and gingival fibroblasts. In addition, the same enzymes were evaluated in gingival samples from healthy and periodontitis-affected individuals.
Primary gingival fibroblast and keratinocyte (HaCaT) cultures were treated with medium containing P. gingivalis LPS or P. gingivalis LPS vehicle for 24 h. After this period, cell viability was assessed by MTT test and total RNA extracted to evaluate gene expression levels of the following enzymes by qRT-PCR: DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a), histone demethylases Jumonji domain containing 3 (JMJD3) and ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX). To evaluate gene expression in healthy and periodontitis-affected individuals, total RNA was extracted from biopsies of gingival tissue from healthy and periodontitis sites, and gene expression of DNMT1, DNAMT3a, JMJD3, and UTX was evaluated by qRT-PCR.
No significant differences were found in the gene expression analysis between healthy and periodontitis-affected gingival samples. The results showed that LPS downregulated DNMT1 (p < 0.05), DNMT3a (p < 0.05), and JMJD3 (p < 0.01) gene expression in HaCaT cells, but no modulation was observed in gingival fibroblasts.
P. gingivalis LPS exposure to human HaCaT keratinocytes downregulates gene expression of the enzymes that promote epigenetic events.
The advance knowledge about epigenetic modifications caused by periodontopathogens may to possibly led to the development of new periodontal therapies.
表观遗传调控在牙周炎等炎症性疾病中的作用知之甚少。本研究旨在评估牙龈卟啉单胞菌脂多糖（LPS）是否可以调节促进人类角质形成细胞和牙龈成纤维细胞中表观遗传事件的一些酶的基因表达水平。此外，还评估了来自健康和牙周炎个体的牙龈样本中的相同酶。
用含有牙龈卟啉单胞菌 LPS 或牙龈卟啉单胞菌 LPS 载体的培养基处理原代牙龈成纤维细胞和角质形成细胞（HaCaT）培养物 24 小时。在此期间，通过 MTT 试验评估细胞活力，并提取总 RNA 以通过 qRT-PCR 评估以下酶的基因表达水平：DNA 甲基转移酶 1（DNMT1）、DNA 甲基转移酶 3a（DNMT3a）、组蛋白去甲基酶 Jumonji 结构域包含 3（JMJD3）和泛在转录四肽重复，X 染色体（UTX）。为了评估健康和牙周炎个体的基因表达，从健康和牙周炎部位的牙龈组织活检中提取总 RNA，并通过 qRT-PCR 评估 DNMT1、DNMT3a、JMJD3 和 UTX 的基因表达。
在健康和牙周炎受影响的牙龈样本的基因表达分析中未发现显着差异。结果表明，LPS 下调了 HaCaT 细胞中 DNMT1（p <0.05）、DNMT3a（p <0.05）和 JMJD3（p <0.01）的基因表达，但在牙龈成纤维细胞中未观察到调节。
牙龈卟啉单胞菌 LPS 暴露于人 HaCaT 角质形成细胞下调促进表观遗传事件的酶的基因表达。
关于牙周病病原体引起的表观遗传修饰的新知识可能导致开发新的牙周治疗方法。
