Anti-HMGB1 Neutralizing Antibody Attenuates Periodontal Inflammation and Bone Resorption in a Murine Periodontitis Model
Anti-HMGB1 Neutralizing Antibody Attenuates Periodontal Inflammation and Bone Resorption in a Murine Periodontitis Model
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DOI：
10.1128/iai.00111-18
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发表时间：
2018-05-01
影响因子：
3.1
通讯作者：
Takashiba, Shogo
中科院分区：
文献类型：
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作者：
Yoshihara-Hirata, Chiaki;Yamashiro, Keisuke;Takashiba, Shogo
高迁移率族蛋白1（HMGB 1）是一种非组蛋白DNA结合蛋白，响应炎症刺激分泌到细胞外环境。分泌的HMGB 1介导各种炎症性疾病，包括牙周炎;然而，HMGB 1诱导牙周炎症的潜在机制尚未完全了解。在这里，我们研究了抗HMGB 1中和抗体是否抑制牙周进展，并在体外和体内研究了HMGB 1的分子病理学。体外分析表明，HMGB 1、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和白细胞介素-1 β（IL-1 β）在用佛波酯肉豆蔻酸酯处理的人牙龈上皮细胞（HGECs）和人单核细胞白血病细胞（THP-1）中响应肿瘤坏死因子-α（TNF-α）刺激而分泌。在体外TNF-α刺激的HGECs和THP-1的条件培养基中观察到GM-CSF和IL-1 β水平增加。TNF-α和抗HMGB 1抗体同时刺激显著降低TNF-α诱导的炎性细胞因子分泌。实验性牙周炎诱导小鼠使用牙龈卟啉单胞菌浸泡结扎。免疫荧光分析证实牙周炎模型小鼠牙龈上皮细胞发生了细胞外易位。全身给予抗HMGB 1中和抗体可显著抑制HMGB 1的易位。抗HMGB 1抗体抑制牙周炎症、IL-1 β和C-X-C基序趋化因子配体1（CXCL 1）的表达、中性粒细胞迁移和骨吸收，通过髓过氧化物酶活性的生物发光成像、定量逆转录-PCR（RT-PCR）和显微计算机断层扫描分析显示。这些发现表明，HMGB 1是响应于牙周感染引起的炎症刺激而分泌的，这对于牙周炎的发生至关重要，并且抗HMGB 1抗体减弱了一系列炎症细胞因子的分泌，从而抑制牙周炎的进展。
High mobility group box 1 (HMGB1) is a non-histone DNA-binding protein that is secreted into the extracellular milieu in response to inflammatory stimuli. The secreted HMGB1 mediates various inflammatory diseases, including periodontitis; however, the underlying mechanisms of HMGB1-induced periodontal inflammation are not completely understood. Here, we examined whether anti-HMGB1 neutralizing antibody inhibits periodontal progression and investigated the molecular pathology of HMGB1 in vitro and in vivo. In vitro analysis indicated that HMGB1, granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-1 beta (IL-1 beta) were secreted in response to tumor necrosis wfactor-alpha (TNF-alpha) stimuli in human gingival epithelial cells (HGECs) and human monocytic leukemia cells (THP-1) treated with phorbol myristate acetate. Increased levels of GM-CSF and IL-1 beta were observed in the conditioned media from TNF-alpha-stimulated HGECs and THP-1 in vitro. Simultaneous stimulation with TNF-alpha and anti-HMGB1 antibody significantly decreased TNF-alpha induced inflammatory cytokine secretion. Experimental periodontitis was induced in mice using Porphyromonas gingivalis-soaked ligatures. The extracellular translocation was confirmed in gingival epithelia in the periodontitis model mice by immunofluorescence analysis. Systemic administration of anti-HMGB1 neutralizing antibody significantly inhibited translocation of HMGB1. The anti-HMGB1 antibody inhibited periodontal inflammation, expression of IL-1 beta and C-X-C motif chemokine ligand 1 (CXCL1), migration of neutrophils, and bone resorption, shown by bioluminescence imaging of myeloperoxidase activity, quantitative reverse transcription-PCR (RT-PCR), and micro-computed tomography analysis. These findings indicate that HMGB1 is secreted in response to inflammatory stimuli caused by periodontal infection, which is crucial for the initiation of periodontitis, and the anti-HMGB1 antibody attenuates the secretion of a series of inflammatory cytokines, consequently suppressing the progression of periodontitis.