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PHF23 negatively regulates the autophagy of chondrocytes in osteoarthritis
- Source :
- Life sciences. 253
- Publication Year :
- 2020
-
Abstract
- Aim Osteoarthritis (OA) is the main cause of disability and joint replacement surgery in the elderly. As a crucial cell survival mechanism, autophagy has been reported to decrease in OA. PHF23 is a new autophagy inhibitor which was first reported by us previously. This study aimed to explore the anti-autophagic mechanism of PHF23 to make it a possible therapeutic target of OA. Main method Lentiviral vectors specific to PHF23 were used on chondrocytes (C28/I2) to establish PHF23 overexpressed or knockdown stable cell strains. Interleukin (IL)-1β (10 ng/mL) and chloroquine (CQ, 25 uM) were used as an inducer of OA and inhibitor of lysosome, respectively. Autophagy was evaluated by autophagosome formation using transmission electron microscopy (TEM) and western blot analysis of P62 and LC3B on different groups of cells. Effects of PHF23 on OA were evaluated by collagen II immunofluorescent staining and western blot analysis of OA-associated proteins MMP13 and ADAMTS5. Effects of PHF23 on AMPK and mTOR/S6K pathways and mitophagy were determined by western blot analysis. Key findings Knockdown of PHF23 enhanced IL-1β-induced autophagy, while overexpression of PHF23 exerted the opposite effect. Knockdown of PHF23 protected chondrocytes against IL-1β-induced OA by decreasing the levels of OA-associated proteins and increasing expression of Collagen II. Knockdown of PHF23 also increased mitophagy level and altered the phosphorylation levels of AMPK, mTOR, and S6K. Significance PHF23 downregulates autophagy, mitophagy in IL-1β-induced OA-like chondrocytes and alters the activities of AMPK and mTOR/S6K, which suggests that PHF23 may be a possible therapeutic target for OA.
- Subjects :
- 0301 basic medicine
Cell Survival
Interleukin-1beta
P70-S6 Kinase 1
AMP-Activated Protein Kinases
030226 pharmacology & pharmacy
General Biochemistry, Genetics and Molecular Biology
03 medical and health sciences
0302 clinical medicine
Chondrocytes
Western blot
Lysosome
Mitophagy
Osteoarthritis
medicine
Autophagy
Humans
General Pharmacology, Toxicology and Pharmaceutics
Collagen Type II
PI3K/AKT/mTOR pathway
Cells, Cultured
Homeodomain Proteins
Gene knockdown
medicine.diagnostic_test
Chemistry
Ribosomal Protein S6 Kinases
TOR Serine-Threonine Kinases
AMPK
General Medicine
Cell biology
030104 developmental biology
medicine.anatomical_structure
Gene Knockdown Techniques
Lysosomes
Subjects
Details
- ISSN :
- 18790631
- Volume :
- 253
- Database :
- OpenAIRE
- Journal :
- Life sciences
- Accession number :
- edsair.doi.dedup.....ace37770a335f7750a9b762f5711fe49