Cyclic stretch promotes the ossification of ligamentum flavum by modulating the Indian hedgehog signaling pathway.
Adult
Aged
Alkaline Phosphatase
/ drug effects
Cell Differentiation
Cells, Cultured
Core Binding Factor Alpha 1 Subunit
/ drug effects
Female
Hedgehog Proteins
/ antagonists & inhibitors
Humans
Ligamentum Flavum
/ metabolism
Male
Middle Aged
Nerve Tissue Proteins
/ metabolism
Nuclear Proteins
/ metabolism
Ossification, Heterotopic
/ genetics
Osteocalcin
/ drug effects
Signal Transduction
/ drug effects
Smoothened Receptor
/ genetics
Sp7 Transcription Factor
/ drug effects
Stress, Mechanical
Veratrum Alkaloids
/ pharmacology
Zinc Finger Protein GLI1
/ metabolism
Zinc Finger Protein Gli2
/ metabolism
Zinc Finger Protein Gli3
/ metabolism
igamentum flavum
cyclic stretch
ossification
osteogenic differentiation
pathway
Indian hedgehog
Journal
Molecular medicine reports
ISSN: 1791-3004
Titre abrégé: Mol Med Rep
Pays: Greece
ID NLM: 101475259
Informations de publication
Date de publication:
08 2020
08 2020
Historique:
received:
22
03
2019
accepted:
29
01
2020
pubmed:
7
7
2020
medline:
24
3
2021
entrez:
7
7
2020
Statut:
ppublish
Résumé
The Indian hedgehog (IHH) signaling pathway is an important pathway for bone growth and development. The aim of the present study was to examine the role of the IHH signaling pathway in the development of the ossification of ligamentum flavum (OLF) at the cellular and tissue levels. The expression levels and localization of the osteogenic genes Runt-related transcription factor 2 (RUNX2), Osterix, alkaline phosphatase (ALP), osteocalcin (OCN) and IHH were evaluated in OLF tissues by reverse transcription-quantitative PCR (RT-qPCR) and immunohistochemistry. Non-ossified ligamentum flavum (LF) sections were used as control samples. The tissue explant method was used to obtain cultured LF cells. In addition, OLF cells were subjected to cyclic stretch application for 0, 6, 12 or 24 h. The expression levels of osteogenic genes, and the IHH signaling pathway genes IHH, Smoothened (SMO), GLI family zinc finger 1 (GLI1), GLI2 and GLI3 were evaluated with RT-qPCR and western blotting. Osteogenic differentiation was further evaluated by assessing ALP activity and staining. Moreover, the effect of cyclopamine (Cpn), an IHH signaling inhibitor, on osteogenic differentiation was examined. The RT-qPCR and immunohistochemical results indicated that the mRNA and protein expression levels of RUNX2, Osterix, ALP, OCN and IHH were significantly higher in the OLF group compared with the LF group. Furthermore, application of cyclic stretch to OLF cells resulted in greater ALP activity, and significant increases in mRNA and protein expression levels of RUNX2, Osterix, ALP and OCN in a time-d00ependent manner. Cyclic stretch application also led to significant increases in IHH signaling pathway genes, including IHH, SMO, GLI1 and GLI2, while no significant effect was found on GLI3 expression level. In addition, it was found that Cpn significantly reversed the effect of cyclic stretch on the ALP activity, and the expression levels of RUNX2, Osterix, ALP, OCN, GLI1 and GLI2. Collectively, the present results suggested that the IHH signaling pathway may mediate the effect of cyclic stretch on the OLF cells.
Identifiants
pubmed: 32626952
doi: 10.3892/mmr.2020.11200
pmc: PMC7339599
doi:
Substances chimiques
BGLAP protein, human
0
Core Binding Factor Alpha 1 Subunit
0
GLI1 protein, human
0
GLI2 protein, human
0
GLI3 protein, human
0
Hedgehog Proteins
0
Nerve Tissue Proteins
0
Nuclear Proteins
0
RUNX2 protein, human
0
SMO protein, human
0
Smoothened Receptor
0
Sp7 Transcription Factor
0
SP7 protein, human
0
Veratrum Alkaloids
0
Zinc Finger Protein GLI1
0
Zinc Finger Protein Gli2
0
Zinc Finger Protein Gli3
0
Osteocalcin
104982-03-8
Alkaline Phosphatase
EC 3.1.3.1
cyclopamine
ZH658AJ192
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1119-1128Références
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