Porphyromonas gingivalis LPS stimulation downregulates DNMT1, DNMT3a, and JMJD3 gene expression levels in human HaCaT keratinocytes
Porphyromonas gingivalis LPS stimulation downregulates DNMT1, DNMT3a, and JMJD3 gene expression levels in human HaCaT keratinocytes
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DOI：
10.1007/s00784-012-0816-z
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发表时间：
2012-08
影响因子：
3.4
通讯作者：
Gláucia Camargo Pereira;G. N. Guimarães;A. C. Planello;M. Santamaria;A. P. Souza;S. Line;M. Marques
中科院分区：
文献类型：
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作者：
Gláucia Camargo Pereira;G. N. Guimarães;A. C. Planello;M. Santamaria;A. P. Souza;S. Line;M. Marques
目的表观遗传调控在牙周炎等炎症性疾病中的作用鲜为人知。本研究的目的是评估牙龈卟啉单胞菌是否能调节一些促进表观遗传事件的酶在培养的人角质形成细胞和牙龈成纤维细胞中的表达水平。此外，在健康和牙周炎患者的牙龈样本中也检测了相同的酶。材料和方法原代培养的牙龈成纤维细胞和角质形成细胞(HaCaT)用含有P的培养液处理。牙周炎是脂多糖酶或牙周炎。用四甲基偶氮唑蓝比色法检测细胞存活率，提取总RNA用qRT-PCR法检测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3(JMJD3)和四肽重复序列X染色体(UTX)的基因表达水平。从健康牙周组织和牙周炎患者牙周组织中提取总RNA，用QRT-PCR法检测DNMT1、DNAMT3a、JMJD3和UTX基因的表达。结果显示，脂多糖可下调HaCaT细胞DNMT1(p<； 0.0 5)、DNMT3a(p<； 0.0 5)和JMJD3(p<； 0.0 1)基因的表达，但对牙周成纤维细胞无明显影响。牙周炎脂多糖暴露于人HaCaT角质形成细胞，下调促进表观遗传事件的酶的基因表达。临床相关性关于牙周病原体引起的表观遗传修饰的高级知识可能会导致新的牙周治疗方法的发展。
ObjectiveThe role of epigenetic regulation in inflammatory diseases such as periodontitis is poorly known. The aim of this study was to assess whetherPorphyromonas gingivalislipopolysaccharide (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.Materials and methodsPrimary gingival fibroblast and keratinocyte (HaCaT) cultures were treated with medium containingP. gingivalisLPS orP. gingivalisLPS 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.ResultsNo 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.ConclusionP. gingivalisLPS exposure to human HaCaT keratinocytes downregulates gene expression of the enzymes that promote epigenetic events.Clinical relevanceThe advance knowledge about epigenetic modifications caused by periodontopathogens may to possibly led to the development of new periodontal therapies.