Activation of vitamin D in the gingival epithelium and its role in gingival inflammation and alveolar bone loss
Activation of vitamin D in the gingival epithelium and its role in gingival inflammation and alveolar bone loss
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DOI：
10.1111/jre.12646
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发表时间：
2019-08-01
影响因子：
3.5
通讯作者：
Diamond, Gill
中科院分区：
文献类型：
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作者：
Menzel, Lorenzo P.;Ruddick, Willam;Diamond, Gill
背景和目的 慢性和侵袭性牙周病都与维生素 D 缺乏有关。维生素 D 的活性形式 1,25(OH)(2)D-3 可诱导培养的人牙龈上皮细胞 (GEC) 中抗菌肽 LL-37 和先天免疫介质的表达。本研究的目的是进一步阐明维生素 D 增强对牙周病 (PD) 发展的先天防御的机制。材料和方法 通过饮食限制使野生型 C57Bl/6 小鼠缺乏维生素 D。用 1,25(OH)(2)D-3 刺激培养的原代和永生化 GEC，然后用牙龈卟啉单胞菌感染，并对活的细胞内细菌进行定量。通过 ELISA 对维生素 D-3 向 25(OH)D-3 和 1,25(OH)(2)D-3 的转化进行定量。通过对野生型小鼠牙龈局部给药，然后进行 qRT-PCR 测定维生素 D 对小鼠基础 IL-1 α 表达的影响。结果在小鼠模型中，限制维生素 D 的饮食会导致牙槽骨流失和牙龈炎症增加。在原代人 GEC 和已建立的人细胞系中，用 1,25(OH)(2)D-3 处理 GEC 可抑制牙龈卟啉单胞菌的细胞内生长。培养的 GEC 表达两种 25-羟化酶（CYP27A1 和 CYP2R1）以及 1-α 羟化酶，能够将维生素 D 转化为 25(OH)D-3 和 1,25(OH)(2)D-3。在小鼠牙龈上局部施用维生素 D-3 和 1,25(OH)(2)D-3 会导致 IL-1 α 表达的快速抑制，IL-1 α 是一种与炎症相关的重要促炎细胞因子，经 RNA-seq 测定，在 1,25(OH)(2)D-3 治疗后，OKF6/TERT1 细胞中 IL-1 α 的表达也比基础水平降低了 2 倍以上。结论 小鼠体内维生素 D 缺乏会导致帕金森病，这与人类的相关性相似，并为研究帕金森病的发展提供了一个独特的模型。维生素 D 可增加 GEC 的活性，抵抗牙周病原体的侵袭，并在体外和体内抑制炎症反应。 GEC 可以将非活性维生素 D 原位转化为活性形式，支持维生素 D 可以直接应用于牙龈以预防或治疗牙周病的假设。
Background and Objective Both chronic and aggressive periodontal disease are associated with vitamin D deficiency. The active form of vitamin D, 1,25(OH)(2)D-3, induces the expression of the antimicrobial peptide LL-37 and innate immune mediators in cultured human gingival epithelial cells (GECs). The aim of this study was to further delineate the mechanism by which vitamin D enhances the innate defense against the development of periodontal disease (PD). Materials and Methods Wild-type C57Bl/6 mice were made deficient in vitamin D by dietary restriction. Cultured primary and immortalized GEC were stimulated with 1,25(OH)(2)D-3, followed by infection with Porphyromonas gingivalis, and viable intracellular bacteria were quantified. Conversion of vitamin D-3 to 25(OH)D-3 and 1,25(OH)(2)D-3 was quantified by ELISA. Effect of vitamin D on basal IL-1 alpha expression in mice was determined by topical administration to the gingiva of wild-type mice, followed by qRT-PCR. Results Dietary restriction of vitamin D led to alveolar bone loss and increased inflammation in the gingiva in the mouse model. In primary human GEC and established human cell lines, treatment of GEC with 1,25(OH)(2)D-3 inhibited the intracellular growth of P. gingivalis. Cultured GEC expressed two 25-hydroxylases (CYP27A1 and CYP2R1), as well as 1-alpha hydroxylase, enabling conversion of vitamin D to both 25(OH)D-3 and 1,25(OH)(2)D-3. Topical application of both vitamin D-3 and 1,25(OH)(2)D-3 to the gingiva of mice led to rapid inhibition of IL-1 alpha expression, a prominent pro-inflammatory cytokine associated with inflammation, which also exhibited more than a 2-fold decrease from basal levels in OKF6/TERT1 cells upon 1,25(OH)(2)D-3 treatment, as determined by RNA-seq. Conclusion Vitamin D deficiency in mice contributes to PD, recapitulating the association seen in humans, and provides a unique model to study the development of PD. Vitamin D increases the activity of GEC against the invasion of periodontal pathogens and inhibits the inflammatory response, both in vitro and in vivo. GEC can convert inactive vitamin D to the active form in situ, supporting the hypothesis that vitamin D can be applied directly to the gingiva to prevent or treat periodontal disease.