Wang Li, He Yanyan, Ning Wanchen
Department of Periodontology, The First Affiliated Hospital of Harbin Medical University, School of Stomatology, Harbin Medical University, Harbin, China.
Department of Stomatology, The First Affiliated Hospital of Zhejiang University, Hangzhou, China.
J Tissue Eng Regen Med. 2021 Jun;15(6):534-545. doi: 10.1002/term.3187. Epub 2021 May 8.
The enhancer of zeste homolog 2 (EZH2) represents a potential target for periodontitis treatment; however, its role in the development of periodontitis remains unclear. The current study aimed to elucidate the role of EZH2 in osteoclasts (OCs) growth as well as the mechanism underpinning the related process. The potential interaction among EZH2, microRNA-101 (miR-101), and vascular cell adhesion molecule 1 (VCAM-1) was evaluated using chromatin immunoprecipitation and dual-luciferase reporter gene assay. The expressions of EZH2 and miR-101 in OCs were examined by Western blot analysis and reverse transcription squantitative polymerase chain reaction. Loss- and gain-function assays were then performed to determine the role of EZH2/miR-101/VCAM-1 in periodontitis and OCs proliferation, followed by OC growth and proliferation detected using tartrate resistant acid phosphatase (TRAP) and 5-ethynyl-2'-deoxyuridine staining. Enzyme-linked immunoassay was conducted to determine the expression of interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α). A periodontitis rat model was established to investigate the effect of EZH2 and VCAM-1 in vivo. EZH2 was overexpressed, while miR-101 was downregulated in the OCs of periodontitis. Silencing of EZH2, VCAM-1 repression, or miR-101 elevation suppressed the growth and proliferation of OC while acting to encumber the release of IL-1β and TNF-α. EZH2 negatively targeted miR-101, while miR-101 negatively targeted VCAM-1. Moreover, silencing of EZH2 or VCAM-1 was observed to attenuate periodontitis which was evidenced by an increase in BMD, BV/TV, and BS/BV as well as reduction in TRAP and cathepsin K in vivo. Taken together, the key findings of the current study demonstrate that EZH2 knockdown inhibited OC formation by elevating the expression of miR-101 via suppression of VCAM-1, ultimately attenuating periodontitis.
zeste同源物2增强子（EZH2）是牙周炎治疗的一个潜在靶点；然而，其在牙周炎发展中的作用仍不清楚。当前研究旨在阐明EZH2在破骨细胞（OC）生长中的作用以及相关过程的潜在机制。使用染色质免疫沉淀和双荧光素酶报告基因检测评估EZH2、微小RNA-101（miR-101）和血管细胞黏附分子1（VCAM-1）之间的潜在相互作用。通过蛋白质免疫印迹分析和逆转录定量聚合酶链反应检测OC中EZH2和miR-101的表达。然后进行功能缺失和功能获得实验，以确定EZH2/miR-101/VCAM-1在牙周炎和OC增殖中的作用，随后使用抗酒石酸酸性磷酸酶（TRAP）和5-乙炔基-2'-脱氧尿苷染色检测OC生长和增殖。进行酶联免疫吸附测定以确定白细胞介素1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的表达。建立牙周炎大鼠模型以研究EZH2和VCAM-1在体内的作用。在牙周炎的OC中，EZH2过表达，而miR-101下调。沉默EZH2、抑制VCAM-1或上调miR-101可抑制OC的生长和增殖，同时抑制IL-1β和TNF-α的释放。EZH2负向靶向miR-101，而miR-101负向靶向VCAM-1。此外，在体内观察到沉默EZH2或VCAM-1可减轻牙周炎，表现为骨密度、骨体积分数和骨表面积/骨体积增加，以及TRAP和组织蛋白酶K减少。综上所述，当前研究的主要发现表明，EZH2基因敲低通过抑制VCAM-1提高miR-101的表达，从而抑制OC形成，最终减轻牙周炎。
