EZH2, a Potential Regulator of Dental Pulp Inflammation and Regeneration
EZH2, a Potential Regulator of Dental Pulp Inflammation and Regeneration
复制标题
EZH2，牙髓炎症和再生的潜在调节剂。
DOI：
10.1016/j.joen.2014.01.031
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发表时间：
2014-08-01
影响因子：
4.2
通讯作者：
Ye, Ling
中科院分区：
文献类型：
--
作者：
Hui, Tianqian;Peng, A.;Ye, Ling
简介：牙髓的再生能力有限，这发生在牙髓炎的早期阶段。存在一种模糊的关系;炎症可能损害或支持牙髓再生。表观遗传学参与细胞增殖和炎症反应，可调控人牙髓细胞（HDPCs）再生。本研究的目的是确定表观遗传标记，增强子zeste同源物2（EZH 2），在炎症，增殖和牙髓再生的作用。我们使用三甲基化组蛋白赖氨酸27（H3 K27 me 3）及其赖氨酸脱甲基酶6 B（KDM 6 B）来监测改变的EZH 2水平的功能性影响。方法：采用免疫组化和免疫荧光法检测牙髓组织中EZH 2、H3 K27 me 3和KDM 6 B等表观遗传标记。通过定量聚合酶链反应和Western blot分析炎症反应或分化过程中HDPC中EZH 2的水平。定量聚合酶链反应用于评估EZH 2抑制对肿瘤坏死因子α刺激后HDPC中白细胞介素的影响。采用细胞计数试剂盒-8、细胞周期和凋亡分析检测细胞增殖。通过定量聚合酶链反应、碱性磷酸酶活性和油红O染色研究HDPC的分化。结果：感染牙髓组织和细胞中EZH 2和H3 K27 me 3表达减少，KDM 6 B表达增加，与HDPC分化相似。EZH 2抑制抑制炎症刺激后HDPC中的IL-1b、IL-6和IL-8信使RNA（mRNA），并通过减少细胞数量、阻滞细胞周期和增加凋亡来阻止HDPC增殖。抑制EZH 2在脂肪形成诱导中损害脂肪形成、过氧化物酶体增殖物激活受体r（PPAR-r）和CCAAT增强子结合蛋白a（CEBP/a）mRNA，而在HDPC矿化诱导中增强碱性磷酸酶活性、Osx和骨唾液酸蛋白（BSP）mRNA。结论：EZH 2抑制HDPC成骨分化，同时增强炎症反应和增殖，表明其在牙髓炎症、增殖和再生中的作用。
Introduction: Dental pulp has limited capability to regenerate, which happens in the early stage of pulpitis. An ambiguous relationship exists; inflammation may impair or support pulp regeneration. Epigenetics, which is involved in cell proliferation and inflammation, could regulate human dental pulp cell (HDPCs) regeneration. The aim of this study was to determine the role of the epigenetic mark, enhancer of zeste homolog 2 (EZH2), in the inflammation, proliferation, and regeneration of dental pulp. We used trimethylated histone lysine 27(H3K27me3) and its lysine demethylase 6B (KDM6B) to monitor functional,effects of altered EZH2 levels. Methods: We detected epigenetic marks (EZH2, H3K27me3, and KDM6B) in pulp tissue by immunohistochemistry and immunofluorescence. EZH2 levels in HDPCs in inflammatory responses or differentiation were analyzed by quantitative polymerase chain reaction and Western blot. Quantitative polymerase chain reaction was used to assess the effects of EZH2 inhibition on interleukins in HDPCs upon tumor necrosis factor alpha stimulation. Cell proliferation was tested by cell counting kit-8, cell cycle, and apoptosis analysis. HDPC differentiation was investigated by quantitative polymerase chain reaction, alkaline phosphatase :activity, and oil red O staining. Results: EZH2 and H3K27me3 were decreased, whereas KDM6B was increased in infected pulp tissue and cells, which were similar to HDPC differentiation. EZH2 inhibition suppressed IL-1b, IL-6, and IL-8 messenger RNA (mRNA) in HDPCs upon inflammatory stimuli and impeded HDPC proliferation by decreasing cell number, arresting cell cycle, and increasing apoptosis. Suppressed EZH2 impaired adipogenesis, peroxisome proliferator-activated receptor r (PPAR-r), and CCAAT-enhancer binding protein a (CEBP/a) mRNA in adipogenic induction while enhancing alkaline phosphatase activity, Osx, and bone sialoprotein (BSP) mRNA in mineralization induction of HDPCs. Conclusions: EZH2 inhibited HDPC osteogenic differentiation while enhancing inflammatory response and proliferation, suggesting its role in pulp inflammation, proliferation, and regeneration.