CRISPR/dCas9-Mediated Parkin Inhibition Impairs Mitophagy and Aggravates Apoptosis of Rat Nucleus Pulposus Cells Under Oxidative Stress
| Autor: | Bin Yan, Tao Lan, Ning-dao Li, Xiao-sheng Chen, Zhe Shen, Yu-chen Zheng |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
TUNEL assay
Chemistry IDD Autophagy apoptosis medicine.disease_cause Biochemistry Genetics and Molecular Biology (miscellaneous) Biochemistry Parkin Cell biology nervous system diseases Intervertebral disk mitophagy Terminal deoxynucleotidyl transferase lcsh:Biology (General) Apoptosis Mitophagy medicine CRISPR/dCas9 Molecular Biosciences lcsh:QH301-705.5 Molecular Biology Oxidative stress Original Research |
| Zdroj: | Frontiers in Molecular Biosciences Frontiers in Molecular Biosciences, Vol 8 (2021) |
| ISSN: | 2296-889X |
| DOI: | 10.3389/fmolb.2021.674632 |
| Popis: | ObjectiveThe aim of this study is to explore the role of Parkin in intervertebral disk degeneration (IDD) and its mitophagy regulation mechanism.Study design and methodsRat nucleus pulposus (NP) cells were stimulated with hydrogen peroxide (H2O2) to a mimic pathological condition. Apoptosis and mitophagy were assessed by Western blot, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and immunofluorescence staining. The CRISPR–dCas9–KRAB system was used to silence the expression of Parkin.ResultIn this study, we found that Parkin was downregulated in rat NP cells under oxidative stress. In addition, treatment with H2O2 resulted in mitochondrial dysfunction, autophagy inhibition, and a significant increase in the rate of apoptosis of NP cells. Meanwhile, mitophagy inhibition enhanced H2O2-induced apoptosis. Furthermore, repression of Parkin significantly attenuated mitophagy and exacerbated apoptosis.ConclusionThese results suggested that Parkin may play a protective role in alleviating the apoptosis of NP cells via mitophagy, and that targeting Parkin may provide a promising therapeutic strategy for the prevention of IDD. |
| Databáze: | OpenAIRE |
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