Related Experiment Video
Updated: Oct 30, 2025
Operational and Intervention Effects of Targeted Tuina in Lumbar Intervertebral Disc Degeneration Model Rabbits
Published on: July 21, 2023
Promoting Nrf2/Sirt3-Dependent Mitophagy Suppresses Apoptosis in Nucleus Pulposus Cells and Protects against
Sunli Hu1,2, Chenxi Zhang1, Tianchen Qian2
1Department of Orthopaedics, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
Abstract:
One of the causes of intervertebral disc degeneration (IVDD) is nucleus pulposus cell (NPC) death, possibly apoptosis. In this study, we explored the role of the Nrf2/Sirt3 pathway and tert-butylhydroquinone (t-BHQ) in IVDD and elucidated the potential working mechanism. Reactive oxygen species (ROS) assay kits and malondialdehyde (MDA) assay kits were used to assess oxidative stress. Western blot and TUNEL staining were used to examine apoptosis. After siRNA against Nrf2 or lentivirus against Sirt3 was transfected into NPCs, the mechanism of the effect of the Nrf2/Sirt3 pathway on NPCs was assessed. The interaction between t-BHQ and its potential interacting protein NRF2 was further investigated through protein docking analysis. ChIP examined the binding affinity between Nrf2 and Sirt3 promoter. In vivo experiments, X-ray, hematoxylin-eosin (HE) staining, Safranin O staining, and immunohistochemistry were used to evaluate IVDD grades. The results demonstrated that activation of the Nrf2/Sirt3 pathway inhibited tert-butyl hydroperoxide- (TBHP-) induced apoptosis and mitochondrial dysfunction in vitro. In addition to apoptosis, upregulation of the Nrf2/Sirt3 pathway induced by t-BHQ restored TBHP-induced autophagic flux disturbances. However, its protective effect was reversed by chloroquine and Si-ATG5. Furthermore, t-BHQ ameliorated IVDD development in a rat model. In conclusion, our findings indicate that the Nrf2/Sirt3 pathway and its agonist represent a potential candidate for treating IVDD.
Insights
Activating the Nrf2/Sirt3 pathway protects nucleus pulposus cells (NPCs) from degeneration by reducing apoptosis and oxidative stress. This pathway, boosted by tert-butylhydroquinone (t-BHQ), shows promise for treating intervertebral disc degeneration (IVDD).
Area of Science:
- Biochemistry
- Cell Biology
- Regenerative Medicine
Background:
- Intervertebral disc degeneration (IVDD) is linked to nucleus pulposus cell (NPC) death, potentially via apoptosis.
- Understanding the molecular mechanisms underlying NPC survival is crucial for developing IVDD treatments.
Purpose of the Study:
- To investigate the role of the Nrf2/Sirt3 pathway in IVDD.
- To explore the therapeutic potential of tert-butylhydroquinone (t-BHQ) in mitigating IVDD.
Main Methods:
- Assessed oxidative stress using ROS and MDA assays.
- Examined apoptosis via Western blot and TUNEL staining.
- Investigated Nrf2/Sirt3 pathway modulation using siRNA and lentivirus transfection, protein docking, and ChIP assays.
- Evaluated IVDD in a rat model using X-ray, HE, Safranin O staining, and immunohistochemistry.
Main Results:
- Activation of the Nrf2/Sirt3 pathway inhibited tert-butyl hydroperoxide (TBHP)-induced apoptosis and mitochondrial dysfunction in NPCs.
- t-BHQ-induced upregulation of Nrf2/Sirt3 restored TBHP-induced autophagic flux disturbances.
- t-BHQ treatment ameliorated IVDD progression in a rat model.
Conclusions:
- The Nrf2/Sirt3 pathway plays a protective role against IVDD by inhibiting NPC apoptosis and mitochondrial dysfunction.
- t-BHQ, an agonist of the Nrf2/Sirt3 pathway, demonstrates therapeutic potential for treating IVDD.
