Changes in DNA Methylation and Chromatin Structure of Pro-inflammatory Cytokines Stimulated by LPS in Broiler Peripheral Blood Mononuclear Cells.
Changes in DNA Methylation and Chromatin Structure of Pro-inflammatory Cytokines Stimulated by LPS in Broiler Peripheral Blood Mononuclear Cells.
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DOI：
10.3382/ps/pew086
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发表时间：
2016-07
期刊：
影响因子：
4.4
通讯作者：
Jing Shen;Yanli Liu;Xiaochun Ren;K. Gao;Yulong Li;Shizhao Li;Junhu Yao;Xiaojun Yang
中科院分区：
文献类型：
--
作者：
Jing Shen;Yanli Liu;Xiaochun Ren;K. Gao;Yulong Li;Shizhao Li;Junhu Yao;Xiaojun Yang
促炎细胞因子IL-1 β、IL-6和肿瘤坏死因子（TNF）-α介导炎症，这是身体的保护性反应，以确保去除有害刺激，以及修复受损组织的愈合过程。促炎细胞因子的过度产生可诱发自身免疫性疾病，并且可能是致命的。本研究旨在探讨脂多糖（LPS）刺激肉鸡外周血单个核细胞（PBMC）后促炎细胞因子表达调控的表观遗传学机制。比较未经处理和LPS处理的PBMC中促炎细胞因子的基因表达、启动子DNA甲基化和染色质可及性。在LPS刺激后测量表观遗传酶DNA甲基转移酶（DNMT）1、组蛋白脱乙酰酶（HDAC）和组蛋白乙酰化酶（HAT）的表达。结果表明，LPS刺激3h后，肉鸡PBMC中促炎细胞因子基因表达被激活。LPS处理后，IL-6基因-302、-264和TNF-α基因-371CpG位点发生去甲基化（P <0.05），而IL-1 β基因启动子区的甲基化模式未受影响。LPS刺激使IL-1 β和IL-6启动子区的染色质结构松弛（P <0.05）。LPS刺激后，DNMT 1和HDAC 2表达降低，p300-CBP相关因子和tat相互作用蛋白-60表达升高（P <0.05）。结果表明，LPS刺激3h后，IL-1 β和IL-6启动子被重塑为可接近的结构，IL-6和TNF-α启动子在特定位点发生去甲基化，这可能影响了促炎细胞因子mRNA的表达。
The pro-inflammatory cytokines IL-1β, IL-6, and tumor necrosis factor (TNF)-α mediate inflammation, which is a protective response by body to ensure removal of detrimental stimuli, as well as a healing process for repairing damaged tissue. The overproduction of pro-inflammatory cytokines can induce autoimmune diseases and can be fatal. The aim of this study was to investigate epigenetic mechanisms in the regulation of pro-inflammatory cytokines expression after lipopolysaccharide (LPS) stimulation of broiler peripheral blood mononuclear cells (PBMC). Gene expression, promoter DNA methylation, and chromatin accessibility of pro-inflammatory cytokines in untreated and LPS-treated PBMC were compared. The expression of epigenetic enzymes DNA methyltransferase (DNMT) 1, histone deacetylase (HDAC), and histone acetylase (HAT) were measured after LPS stimulation. The results showed the activated gene expression of pro-inflammatory cytokines in broiler PBMC stimulated 3 h by LPS. The demethylation of IL-6 gene - 302 and -264 cytosine-guanine (CpG) sites, as well as TNF-α gene -371 CpG site, occurred after LPS treatment (P < 0.05), whereas the methylaiton pattern in the IL-1β gene promoter region was not affected. Otherwise, LPS stimulation relaxed the chromatin structure at IL-1β and IL-6 promoter (P < 0.05). The lower expression of DNMT1 and HDAC2, and higher expression of p300-CBP-associated factor and tat-interaction protein-60, were detected in response to LPS (P < 0.05). Our data indicated that after LPS stimulation for 3 h, IL-1β and IL-6 promoter are remodeled into an accessible structure, and the IL-6 and TNF-α promoter are demethylated at special sites, which possible impact the mRNA expression of pro-inflammatory cytokines.