Protein Kinase C and Matrix Metalloproteinases Expression Using Phorbol Myristate Acetate in Degenerative Intervertebral Disc Cells.
Humans
Intervertebral Disc Degeneration
/ metabolism
Tetradecanoylphorbol Acetate
/ pharmacology
Protein Kinase C
/ metabolism
Nucleus Pulposus
/ metabolism
Adult
Middle Aged
Female
Male
Wnt Signaling Pathway
/ drug effects
Cells, Cultured
beta Catenin
/ metabolism
Tissue Inhibitor of Metalloproteinase-1
/ metabolism
TRPV Cation Channels
/ metabolism
Matrix Metalloproteinases
/ metabolism
Interleukin-6
/ metabolism
ADAMTS5 Protein
/ metabolism
Matrix Metalloproteinase 13
/ metabolism
Matrix Metalloproteinase 1
/ metabolism
Disc degeneration
Matrix metalloproteinase
Nucleus pulposus
Phorbol myristate acetate
Protein kinase C-delta
Journal
Clinics in orthopedic surgery
ISSN: 2005-4408
Titre abrégé: Clin Orthop Surg
Pays: Korea (South)
ID NLM: 101505087
Informations de publication
Date de publication:
Oct 2024
Oct 2024
Historique:
received:
14
11
2023
revised:
30
03
2024
accepted:
15
05
2024
medline:
4
10
2024
pubmed:
4
10
2024
entrez:
4
10
2024
Statut:
ppublish
Résumé
Degeneration of nucleus pulposus (NP) cells involves multiple factors. The relationship between the canonical Wnt/β-catenin signaling pathway and matrix metalloproteinases (MMPs) is important in cellular senescence. Protein kinase C (PKC), an intermediate of the non-canonical Wnt pathway stimulated by phorbol myristate acetate (PMA), possibly prevents NP cell senescence, although not yet demonstrated in human-based studies. This study aimed to investigate the effect of PMA stimulation on the non-canonical and canonical Wnt pathways and MMP expression in human NP cells to ascertain its inhibitory effects on the senescence of NP cells. Human disc tissues of Pfirrmann grades 1 and 2 were collected from patients during spinal surgery and subsequently cultured. Protein and ribonucleic acid (RNA) were isolated from NP cells treated with PMA (400 nM) for 24 hours. Expression of MMP1, MMP13, tissue inhibitor of matrix metalloproteinase 1 (TIMP1), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), transient receptor potential vanilloid 4 (TRPV4), interleukin-6 (IL-6), and β-catenin were detected using western blot analysis. Messenger RNA (mRNA) expression of type II collagen and glycosaminoglycan (GAG) were analyzed using reverse transcription polymerase chain reaction. IL-6 and prostaglandin E2 (PGE Expression of PKC-δ (intermediate of the non-canonical Wnt pathway) and β-catenin (intermediate of the canonical Wnt pathway) was increased by PMA treatment. The mRNA levels of type II collagen and GAG increased; however, their protein levels were not altered. PMA treatment increased the expression of MMP1, TIMP1, ADAMTS5, IL-6, PGE PMA activated PKC-δ, affecting the non-canonical Wnt pathway; however, its effect on β-catenin in the canonical Wnt pathway was limited. β-catenin activation through the TRPV4 channel led to increased expression of MMP1 and ADAMTS5 and that of IL-6 and PGE
Sections du résumé
Background
UNASSIGNED
Degeneration of nucleus pulposus (NP) cells involves multiple factors. The relationship between the canonical Wnt/β-catenin signaling pathway and matrix metalloproteinases (MMPs) is important in cellular senescence. Protein kinase C (PKC), an intermediate of the non-canonical Wnt pathway stimulated by phorbol myristate acetate (PMA), possibly prevents NP cell senescence, although not yet demonstrated in human-based studies. This study aimed to investigate the effect of PMA stimulation on the non-canonical and canonical Wnt pathways and MMP expression in human NP cells to ascertain its inhibitory effects on the senescence of NP cells.
Methods
UNASSIGNED
Human disc tissues of Pfirrmann grades 1 and 2 were collected from patients during spinal surgery and subsequently cultured. Protein and ribonucleic acid (RNA) were isolated from NP cells treated with PMA (400 nM) for 24 hours. Expression of MMP1, MMP13, tissue inhibitor of matrix metalloproteinase 1 (TIMP1), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), transient receptor potential vanilloid 4 (TRPV4), interleukin-6 (IL-6), and β-catenin were detected using western blot analysis. Messenger RNA (mRNA) expression of type II collagen and glycosaminoglycan (GAG) were analyzed using reverse transcription polymerase chain reaction. IL-6 and prostaglandin E2 (PGE
Results
UNASSIGNED
Expression of PKC-δ (intermediate of the non-canonical Wnt pathway) and β-catenin (intermediate of the canonical Wnt pathway) was increased by PMA treatment. The mRNA levels of type II collagen and GAG increased; however, their protein levels were not altered. PMA treatment increased the expression of MMP1, TIMP1, ADAMTS5, IL-6, PGE
Conclusions
UNASSIGNED
PMA activated PKC-δ, affecting the non-canonical Wnt pathway; however, its effect on β-catenin in the canonical Wnt pathway was limited. β-catenin activation through the TRPV4 channel led to increased expression of MMP1 and ADAMTS5 and that of IL-6 and PGE
Identifiants
pubmed: 39364116
doi: 10.4055/cios23365
pmc: PMC11444957
doi:
Substances chimiques
Tetradecanoylphorbol Acetate
NI40JAQ945
Protein Kinase C
EC 2.7.11.13
beta Catenin
0
Tissue Inhibitor of Metalloproteinase-1
0
TRPV Cation Channels
0
Matrix Metalloproteinases
EC 3.4.24.-
Interleukin-6
0
ADAMTS5 Protein
EC 3.4.24.-
Matrix Metalloproteinase 13
EC 3.4.24.-
Matrix Metalloproteinase 1
EC 3.4.24.7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
827-835Informations de copyright
Copyright © 2024 by The Korean Orthopaedic Association.
Déclaration de conflit d'intérêts
CONFLICT OF INTEREST: No potential conflict of interest relevant to this article was reported.