Hypoxia stimulates collagen hydroxylation in gingival fibroblasts and periodontal ligament cells
Autor: | Kazuma Kawakami, Chiaki Morimoto, Shinya Murakami, Keigo Sawada, Masahide Takedachi, Tomoaki Iwayama, Kohsuke Kawasaki, Junpei Shimomura, Mari Murata, Asae Hirai |
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Rok vydání: | 2021 |
Předmět: |
0301 basic medicine
Periodontal Ligament Angiogenesis Hydroxylation Extracellular matrix 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Gene expression medicine Humans Hypoxia Fibroblast Cells Cultured Tissue homeostasis Chemistry 030206 dentistry Transfection Fibroblasts Hypoxia-Inducible Factor 1 alpha Subunit Molecular biology Cell Hypoxia 030104 developmental biology medicine.anatomical_structure Periodontics Collagen Type I collagen |
Zdroj: | Journal of Periodontology. 92:1635-1645 |
ISSN: | 1943-3670 0022-3492 |
Popis: | Background Cellular responses to hypoxia regulate various biological events, including angiogenesis and extracellular matrix metabolism. Collagen is a major component of the extracellular matrix in periodontal tissues and its coordinated production is essential for tissue homeostasis. In this study, we investigated the effects of hypoxia on collagen production in human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLs). Methods HGFs and HPDLs were cultured under either normoxic (20% O2 ) or hypoxic (1% O2 ) conditions. Nuclear expression of hypoxia-inducible factor-1α (HIF-1α) was determined by western blotting. Peri-cellular expression of type I collagen was examined by immunocytochemistry analysis. Synthesis of type I collagen was evaluated by measuring the concentration of procollagen type I C-peptide (PIP) in culture supernatant using enzyme-linked immunosorbent assay. Expression of collagen hydroxylase enzymes prolyl 4-hydroxylase alpha polypeptide 1 (P4HA1) and 2-oxoglutarate 5-dioxygenase 2 (PLOD2) was determined by RT-qPCR and western blotting. The roles of these enzymes were analyzed using siRNA transfection. Results Cultivation under hypoxic conditions stimulated type I collagen production via HIF-1α in both cell types. Interestingly, hypoxic conditions did not affect collagen 1a1 or 1a2 gene expression but upregulated that of P4HA1 and PLOD2. Additionally, suppressing P4HA1 significantly decreased the levels of hypoxia-induced procollagen type I C-peptide, a product of stable triple helical collagen, in the supernatant. In contrast, PLOD2 suppression decreased cross-linked collagen expression in the pericellular region. Conclusion Our results suggest that hypoxia activates collagen synthesis in HGFs and HPDLs by upregulating hydroxylases P4HA1 and PLOD2 in an HIF-1α-dependent manner. |
Databáze: | OpenAIRE |
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