College of Dentistry, Department of Periodontology, University of Tennessee Health Science Center, Memphis, TN, USA.
Department of Bioscience Research, University of Tennessee Health Science Center, Memphis, TN, USA.
J Periodontal Res. 2017 Oct;52(5):832-841. doi: 10.1111/jre.12452. Epub 2017 Mar 27.
Vitamin D-1,25(OH) D or 1,25D3-maintains healthy osseous tissue, stimulates the production of the antimicrobial peptide cathelicidin and has anti-inflammatory effects, but it can cause hypercalcemia. Evidence links diminished serum levels of 1,25D3 with increased gingival inflammation. Periodontitis progression is associated with increased local production of inflammatory mediators by immune cells and gingival fibroblasts. These include interleukin (IL)-6, a regulator of osteoclastic bone resorption, and the neutrophil chemoattractant IL-8, both regulated by signaling pathways, including NF-κB and MAPK/AP-1. The objectives were to determine the effects of 1,25D3 or a non-calcemic analog, 20-hydroxyvitamin D -20(OH)D or 20D3-on IL-1β-stimulated IL-6 and IL-8 production, and NF-κB and MAPK/AP-1 activation, by human gingival fibroblasts.
Human gingival fibroblasts were incubated ± IL-1β, with or without exposure to 1,25D3 or 20D3. IL-6 and IL-8 in culture supernatants were measured by enzyme-linked immunosorbent assay. NF-κB (p65) and AP-1 (phospho-cJun) and were measured in nuclear extracts via binding to specific oligonucleotides. Data were analyzed using ANOVA and Scheffe's F procedure for post hoc comparisons.
IL-1β-stimulated IL-6 and IL-8 levels were both significantly inhibited (40%-60%) (P<.045) by 1,25D3, but not 20D3 (0%-15% inhibition, not statistically significant). Both 1,25D3 and 20D3 significantly and similarly inhibited IL-1β-stimulated nuclear levels of p65 and phospho-cJun (P<.02).
Reduction of the activation of NF-κB and AP-1 alone is not able to inhibit strongly the IL-1β stimulated IL-6 and IL-8 gene expression. 1,25D3 but not 20D3 may affect some of the many other factors/processes/pathways that in turn regulate the expression of these genes. However, the results suggest that topical application of ligands of the vitamin D receptor may be useful in the local treatment of periodontitis while reducing adverse systemic effects.
维生素 D-1,25(OH)D 或 1,25D3 可维持健康的骨骼组织，刺激抗菌肽 cathelicidin 的产生，并具有抗炎作用，但可能导致高钙血症。有证据表明，血清 1,25D3 水平降低与牙龈炎症增加有关。牙周炎的进展与免疫细胞和牙龈成纤维细胞产生的局部炎症介质增加有关。这些炎症介质包括白细胞介素 (IL)-6，一种破骨细胞骨吸收的调节剂，以及中性粒细胞趋化因子 IL-8，两者均受信号通路调节，包括 NF-κB 和 MAPK/AP-1。目的是确定 1,25D3 或非钙类似物 20-羟基维生素 D-20(OH)D 或 20D3 对人牙龈成纤维细胞 IL-1β 刺激的 IL-6 和 IL-8 产生以及 NF-κB 和 MAPK/AP-1 激活的影响。
将人牙龈成纤维细胞孵育 ±IL-1β，同时或不暴露于 1,25D3 或 20D3。通过酶联免疫吸附试验测量培养上清液中的 IL-6 和 IL-8。通过与特定寡核苷酸结合，测量核提取物中的 NF-κB（p65）和 AP-1（磷酸化 cJun）。使用方差分析和 Scheffe 的 F 程序进行事后比较来分析数据。
1,25D3 显著抑制 (40%-60%) (P<.045) IL-1β 刺激的 IL-6 和 IL-8 水平，但 20D3 则没有 (0%-15%抑制，无统计学意义)。1,25D3 和 20D3 均显著且相似地抑制了 IL-1β 刺激的核 p65 和磷酸化 cJun 的水平 (P<.02)。
NF-κB 和 AP-1 的激活减少单独不足以强烈抑制 IL-1β 刺激的 IL-6 和 IL-8 基因表达。1,25D3 但不是 20D3 可能会影响许多其他因素/过程/途径，这些因素/过程/途径反过来又调节这些基因的表达。然而，结果表明，维生素 D 受体配体的局部应用可能有助于局部治疗牙周炎，同时减少不良的全身影响。
