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Hsa_circ_0001946 Ameliorates Mechanical Stress-induced Intervertebral Disk Degeneration Via Targeting miR-432-5p and
Qian Xiang1, Juntan Wang, Zhangrong Cheng
1Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
Study Design:
Experimental analysis of circular RNA in intervertebral disk degeneration (IDD).
Objective:
This study aimed to explore the roles of hsa_circ_0001946 (circ-CDR1as) in mechanical stress-induced nucleus pulposus cell injury in IDD.
Summary Of Background Data:
Mechanical stress is an important pathogenic factor for IDD. Excessive compression stress leads to nucleus pulposus (NP) cell apoptosis and extracellular matrix (ECM) degradation and accelerated IDD. Circ-CDR1as is associated with various degenerative conditions, but its role in IDD is not clear. Herein, we explored the roles and mechanisms of circ-CDR1as in IDD in vitro.
Materials And Methods:
An in vitro model of IDD was constructed by treating NP cells with 1.0 MPa compression stress. Quantitative real-time polymerase chain reaction assay was used for detecting the expression of circ-CDR1as and miR-432-5p. Immunofluorescent analysis was performed for MMP13 detection. Western blot assay was performed for detecting apoptosis and ECM-related protein expression. Flow cytometry analysis was used for cell apoptosis analysis. The dual-luciferase reporter was used to analyze the interaction between miR-432-5p and circ-CDR1as or SOX9. Differences in means between groups were evaluated using the Student t test or one-way analysis of variance.
Results:
In compression-treated human NP cells, we found that circ-CDR1as was significantly downregulated. Functional experiments showed that circ-CDR1as overexpression reduced the compression-induced apoptosis and ECM degradation in NP cells. Further research indicated that circ-CDR1as could act as a molecular sponge for miR-432-5p, a miRNA that enhanced compression-induced damage of NP cells by inhibiting the expression of SOX9. The luciferase reporter experiments also showed that the mutual dialogue between circ-CDR1as and miR-432-5p regulated the expression of SOX9.
Conclusions:
Circ-CDR1as binds to miR-432-5p and plays a protective role in mitigating compression-induced NP cell apoptosis and ECM degradation by targeting SOX9. Circ-CDR1as may provide a novel therapeutic target for the clinical management of IDD in the future.
Insights
Circular RNA circ-CDR1as protects against intervertebral disc degeneration (IDD) by reducing nucleus pulposus cell apoptosis and extracellular matrix degradation. This finding offers a potential therapeutic target for IDD.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Intervertebral disc degeneration (IDD) is a degenerative condition exacerbated by mechanical stress.
- Compression stress induces nucleus pulposus (NP) cell apoptosis and extracellular matrix (ECM) degradation, key factors in IDD.
- The role of circular RNA (circ-CDR1as) in IDD pathogenesis remains unclear.
Purpose of the Study:
- To investigate the role and mechanism of hsa_circ_0001946 (circ-CDR1as) in mechanical stress-induced nucleus pulposus cell injury in IDD.
- To explore the potential of circ-CDR1as as a therapeutic target for IDD.
Main Methods:
- An in vitro IDD model was established using NP cells subjected to compression stress.
- Quantitative real-time PCR, immunofluorescence, Western blot, and flow cytometry were employed to assess molecular and cellular changes.
- Dual-luciferase reporter assays were used to elucidate the interaction between circ-CDR1as, miR-432-5p, and SOX9.
Main Results:
- Circ-CDR1as expression was significantly downregulated in NP cells under compression stress.
- Overexpression of circ-CDR1as attenuated compression-induced apoptosis and ECM degradation in NP cells.
- Circ-CDR1as acted as a molecular sponge for miR-432-5p, inhibiting its suppressive effect on SOX9 expression and thereby mitigating cellular damage.
Conclusions:
- Circ-CDR1as plays a protective role in IDD by inhibiting NP cell apoptosis and ECM degradation via the circ-CDR1as/miR-432-5p/SOX9 axis.
- Circ-CDR1as represents a promising therapeutic target for managing intervertebral disc degeneration.
