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牙周组织细胞中维生素 D 通路的初步研究。

Preliminary investigation of the vitamin D pathway in periodontal connective tissue cells.

机构信息

Department of Periodontology, Peking University School and Hospital of Stomatology, Beijing, China.

Department of Stomatology, Peking University Third Hospital, Beijing, China.

出版信息

J Periodontol. 2018 Mar;89(3):294-302. doi: 10.1002/JPER.17-0530. Epub 2018 Mar 12.

Abstract

BACKGROUND

The vitamin D pathway, from toll-like receptor activation to human cationic antimicrobial protein (hCAP-18/LL-37) generation, has been identified in monocytes and keratinocytes. This study aimed to investigate the vitamin D pathway in human gingival fibroblasts (hGFs) and human periodontal ligament cells (hPDLCs) and to provide preliminary evidence of its role in periodontal immune defense.

METHODS

Primary cultures of hGFs and hPDLCs were stimulated with 1,25-dihydroxy vitamin D and 25-hydroxy vitamin D , with or without Porphyromonas gingivalis lipopolysaccharide. CYP27B1 RNA interference and vitamin D receptor (VDR) antagonism were also used for reverse proof. The mRNA expression of hCAP-18/LL-37, VDR, interleukin (IL)-6, IL-8, and monocyte chemotactic protein-1 were detected using real-time polymerase chain reaction. The LL-37 concentrations were measured using enzyme-linked immunosorbent assay.

RESULTS

In hGFs and hPDLCs, 25-hydroxy vitamin D and 1,25-dihydroxy vitamin D induced hCAP-18/LL-37 expression, which was further increased by Porphyromonas gingivalis lipopolysaccharide. If the function of CYP27B1 or VDR was blocked, the induction was significantly weakened. IL-8 and monocyte chemotactic protein-1 mRNA expression could be suppressed by the vitamin D pathway.

CONCLUSION

These findings suggest that the vitamin D pathway exists in hGFs and hPDLCs and plays an important role in immune defense in periodontal soft tissues.

摘要

背景

从 Toll 样受体激活到人类阳离子抗菌蛋白(hCAP-18/LL-37)的生成,维生素 D 途径已在单核细胞和角质形成细胞中被鉴定出来。本研究旨在探讨人牙龈成纤维细胞(hGFs)和人牙周韧带细胞(hPDLCs)中的维生素 D 途径,并为其在牙周免疫防御中的作用提供初步证据。

方法

用 1,25-二羟维生素 D 和 25-羟维生素 D 对原代培养的 hGFs 和 hPDLCs 进行刺激,同时或不加入牙龈卟啉单胞菌脂多糖。还使用 CYP27B1 RNA 干扰和维生素 D 受体(VDR)拮抗剂进行反向验证。采用实时聚合酶链反应检测 hCAP-18/LL-37、VDR、白细胞介素(IL)-6、IL-8 和单核细胞趋化蛋白-1 的 mRNA 表达。采用酶联免疫吸附试验检测 LL-37 浓度。

结果

在 hGFs 和 hPDLCs 中,25-羟维生素 D 和 1,25-二羟维生素 D 诱导 hCAP-18/LL-37 表达,牙龈卟啉单胞菌脂多糖进一步增加了其表达。如果阻断 CYP27B1 或 VDR 的功能,诱导作用明显减弱。维生素 D 途径可抑制 IL-8 和单核细胞趋化蛋白-1 mRNA 的表达。

结论

这些发现表明,维生素 D 途径存在于 hGFs 和 hPDLCs 中,在牙周软组织的免疫防御中发挥重要作用。

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