Department of Periodontology, Peking University School and Hospital of Stomatology, Beijing, China.
National Clinical Research Center for Oral Diseases, Beijing, China.
J Periodontol. 2021 Jul;92(7):44-53. doi: 10.1002/JPER.20-0225. Epub 2020 Nov 10.
In periodontal connective tissue cells, the vitamin D pathway has been elucidated, and vitamin D in the main storage form, 25-hydroxy vitamin D (25[OH]D ), and the functional form, 1,25-dihydroxy vitamin D (1,25[OH] D ), have been found to induce the expression of human cationic antimicrobial protein (hCAP-18)/LL-37. Moreover, synergistic effects between Toll-like receptor agonists and 25(OH)D have been reported. This research aimed at extending the vitamin D pathway to vitamin D and CYP27A1 in human periodontal ligament cells (hPDLCs) to further explore its function in periodontal inflammatory reaction.
Vitamin D was used to stimulate hPDLCs in the presence or absence of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS). Conversely, CYP27A1 RNA interference was performed to further validate the findings. The mRNA expression of hCAP-18 was determined with real-time polymerase chain reaction. Monocyte chemotactic protein-1 (MCP-1) and interleukin-8 (IL-8) were also detected. The cell supernatant levels of LL-37 were detected with enzyme-linked immunosorbent assay.
Vitamin D significantly enhanced the generation of hCAP-18/LL-37. A combination of Pg-LPS and vitamin D significantly promoted hCAP-18/LL-37 expression. When the expression of CYP27A1 was knocked down with RNA interference, the induction of hCAP-18/LL-37 expression was significantly inhibited. Therefore, the mRNA levels of MCP-1 and IL-8 in hPDLCs were significantly decreased through the vitamin D pathway.
The vitamin D pathway from vitamin D to hCAP-18/LL-37 exists in hPDLCs, and CYP27A1 might be involved in periodontal immune defense.
在牙周结缔组织细胞中,维生素 D 通路已被阐明,维生素 D 的主要储存形式 25-羟维生素 D(25[OH]D)和功能形式 1,25-二羟维生素 D(1,25[OH]D)被发现可诱导人阳离子抗菌蛋白(hCAP-18)/LL-37 的表达。此外,已报道 Toll 样受体激动剂与 25(OH)D 之间存在协同作用。本研究旨在将维生素 D 通路扩展到人牙周韧带细胞(hPDLC)中的维生素 D 和 CYP27A1,以进一步探讨其在牙周炎症反应中的作用。
用维生素 D 刺激存在或不存在牙龈卟啉单胞菌脂多糖(Pg-LPS)的 hPDLCs。相反,进行 CYP27A1 RNA 干扰以进一步验证发现。用实时聚合酶链反应测定 hCAP-18 的 mRNA 表达。还检测单核细胞趋化蛋白-1(MCP-1)和白细胞介素-8(IL-8)。用酶联免疫吸附试验检测 LL-37 的细胞上清液水平。
维生素 D 显著增强 hCAP-18/LL-37 的产生。Pg-LPS 和维生素 D 的组合显著促进 hCAP-18/LL-37 的表达。用 RNA 干扰敲低 CYP27A1 的表达时,hCAP-18/LL-37 的诱导表达显著受到抑制。因此,hPDLCs 中 MCP-1 和 IL-8 的 mRNA 水平通过维生素 D 通路显著降低。
hPDLCs 中存在从维生素 D 到 hCAP-18/LL-37 的维生素 D 通路,CYP27A1 可能参与牙周免疫防御。