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Updated: Oct 16, 2025
Software-Assisted Quantitative Measurement of Osteoarthritic Subchondral Bone Thickness
Published on: March 18, 2022
MiR-4303 relieves chondrocyte inflammation by targeting ASPN in osteoarthritis
Chunyu Wang1, Li Wang2, Xingfa Guan3
1Department of Orthopedics, Chifeng Municipal Hospital, Chifeng, 024000, Inner Mongolia Autonomous Region, China.
Background:
Osteoarthritis (OA) is a severe articular cartilage disease whose pathogenesis involves the inflammation of chondrocytes. MicroRNAs (miRNAs) are considered to be effective inflammation regulators. However, the regulatory mechanism of miRNAs in osteoarthritis needs to be further elucidated. In this paper, we aim to investigate the underlying mechanisms by which miR-4303 regulates osteoarthritis.
Methods:
RT-qPCR is performed to detect the mRNA expression levels of miR-4303, ASPN, PDIA3, PIK3CA, and TRAF3. CCK-8 assay and EdU assay are carried to assess chondrocyte viability. The protein expression levels of ASPN, PCNA, Ki-67, CyclinA1, CyclinB1, CyclinD2, p27, Bax, Bcl-2, cleaved caspase-3, and Cleaved caspase-9 were measured by western blot. FACs is performed to detect the cell cycle and apoptosis of chondrocyte. ELISA is conducted to assess the levels of TNF-β, IL-1β and IL-6 in the supernatant of chondrocytes. The potential binding sites of miR-4303 and ASPN are predicted by the miRDB database and confirmed by the dual-luciferase reporter gene assay.
Results:
Our findings illustrated that miR-4303 was down-regulated in arthritic tissues and LPS-induced chondrocytes; miR-4303 overexpression rescued the decrease in cell viability, cell cycle arrest and apoptosis induced by LPS. Furthermore, miR-4303 overexpression inhibited the release of inflammatory factors in LPS-induced chondrocytes, miR-4303 relieved chondrocyte inflammation via targeting ASPN.
Conclusion:
MiR-4303 serves as a prognostic biomarker and relieves chondrocyte inflammation via targeting ASPN. Our findings provide novel prognostic biomarkers in predicting the progression and prognosis of osteoarthritis.
Insights
MicroRNA-4303 (miR-4303) is downregulated in osteoarthritis and protects chondrocytes from inflammation and apoptosis by targeting ASPN. This finding offers new prognostic biomarkers for osteoarthritis.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Osteoarthritis (OA) is a degenerative joint disease characterized by articular cartilage breakdown and chondrocyte inflammation.
- MicroRNAs (miRNAs) play crucial roles in regulating cellular inflammation, but their specific mechanisms in OA pathogenesis remain unclear.
- Investigating the role of specific miRNAs, such as miR-4303, is essential for understanding OA progression.
Purpose of the Study:
- To elucidate the regulatory mechanisms of miR-4303 in osteoarthritis.
- To determine if miR-4303 influences chondrocyte viability, cell cycle, apoptosis, and inflammation.
- To identify the downstream targets of miR-4303 involved in OA pathogenesis.
Main Methods:
- Quantitative real-time PCR (RT-qPCR) to measure miRNA and mRNA expression.
- Cell viability assays (CCK-8, EdU) and western blotting for protein analysis.
- Flow cytometry (FACs) for cell cycle and apoptosis analysis, and ELISA for inflammatory cytokine levels.
- Bioinformatic prediction (miRDB) and dual-luciferase reporter assay to confirm miRNA-target interaction.
Main Results:
- miR-4303 expression was significantly downregulated in osteoarthritis tissues and lipopolysaccharide (LPS)-induced chondrocytes.
- Overexpression of miR-4303 reversed LPS-induced decreases in chondrocyte viability, cell cycle arrest, and apoptosis.
- miR-4303 overexpression suppressed the release of pro-inflammatory cytokines (TNF-β, IL-1β, IL-6) and targeted ASPN to alleviate chondrocyte inflammation.
Conclusions:
- miR-4303 acts as a protective factor in osteoarthritis by inhibiting chondrocyte inflammation.
- ASPN is identified as a direct target of miR-4303, mediating its anti-inflammatory effects.
- miR-4303 demonstrates potential as a novel prognostic biomarker for osteoarthritis progression and prognosis.
