RRC ID 58651
著者 Nakanishi J, Suzuki S, Yoshida K, Hirata-Tsuchiya S, Haruyama N, Yamada S, Shiba H.
タイトル Dentin phosphoprotein inhibits lipopolysaccharide-induced macrophage activation independent of its serine/aspartic acid-rich repeats.
ジャーナル Arch Oral Biol
Abstract OBJECTIVE:The objective of this study was to investigate the effects of dentin phosphoprotein (DPP) on lipopolysaccharide-induced inflammatory responses of macrophages in vitro.
DESIGN:Wildtype and mutant recombinant dentin phosphoprotein (rDPP) proteins were generated using a mammalian expression system. Macrophages, phorbol 12-myristate 13-acetate-differentiated THP-1 cells, were stimulated with lipopolysaccharide in the absence or presence of rDPP proteins. After the 24-hr incubation, the inflammatory gene expression levels were examined by quantitative reverse-transcription polymerase chain reaction and the amount of secreted TNF-α protein was evaluated by enzyme-linked immunosorbent assay. Furthermore, the subcellular localization of exogenously added rDPP was examined by immunocytochemistry, and the direct binding of rDPP to lipopolysaccharide was quantified by solid-phase binding assay.
RESULTS:rDPP dose-dependently reduced the expression of lipopolysaccharide-induced inflammatory genes, such as TNFα, IL-1β, and IL-8, and TNF-α protein secretion from the macrophages. Furthermore, mutant rDPP having a shortened serine/aspartic acid-rich repeats (SDrr) was also able to inhibit lipopolysaccharide-induced inflammatory responses of macrophages. rDPP was localized adjacent to the cellular membrane rather than in the cytoplasm, and rDPP was able to bind to lipopolysaccharide. These results suggested that rDPP inhibited lipopolysaccharide-induced inflammatory responses by binding to lipopolysaccharide.
CONCLUSIONS:In addition to the well-known functions of DPP for dentin mineralization that depend on the SDrr, we demonstrated that DPP possesses anti-inflammatory effects on lipopolysaccharide-stimulated macrophages that are independent of the SDrr.
巻・号 110
ページ 104634
公開日 2020-2-1
DOI 10.1016/j.archoralbio.2019.104634
PII S0003-9969(19)30367-X
PMID 31855746
MeSH Animals Aspartic Acid Dentin* / immunology Extracellular Matrix Proteins* / pharmacology Inflammation Lipopolysaccharides Macrophage Activation* Phosphoproteins* / pharmacology Serine Sialoglycoproteins* / pharmacology Tumor Necrosis Factor-alpha
IF 1.931
引用数 0
リソース情報
ヒト・動物細胞 THP-1