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MiR-20a-5p Regulates MPP+-Induced Oxidative Stress and Neuroinflammation in HT22 Cells by Targeting IRF9/NF-κB Axis
Qiang Wang1, Yuan Wang1, Feng Zhou2
1College of Acu-Moxibustion and Massage, Shaanxi University of Chinese Medicine, Xianyang 712046, Shaanxi, China.
Abstract:
Substantial evidence indicates that microRNAs (miRNAs) can be used as biological markers of Parkinson's disease (PD) and contribute to the risk assessment, early diagnosis, and treatment. We aimed to explore the role and potential mechanism of miR-20a-5p on inflammation and oxidative stress in 1-methyl-4-phenyl pyridine ion- (MPP+-) induced HT22 cells. HT22 cells were pretreated with miR-20a-5p mimic and/or pcDNA-IRF9 for 24 h and then treated with MPP+ (0.5 mM) for 24 h. The cell viability and apoptosis were determined using the Cell Counting Kit-8 (CCK-8) and Annexin V FITC/PI staining flow cytometry assay, respectively. The expression and secretion of inflammatory factors and oxidative stress-related factors were detected by enzyme-linked immunosorbent assay (ELISA). The protein expression levels were detected using Western blot analysis. Here, we discovered that MPP+ led to mitochondrial dysfunction, inflammation, and cell damage of HT22 cells, which were alleviated by miR-20a-5p overexpression. We further clarified that interferon regulatory factor 9 (IRF9) was a target gene of miR-20a-5p. IRF9 contributed to MPP+-induced mitochondrial disruption, inflammation, and cell apoptosis. Moreover, IRF9 hindered the improvement of miR-20a-5p overexpression on MPP+-induced neurotoxicity. Furthermore, the decrease of p-P65 level induced by miR-20a-5p mimic was significantly reversed by IRF9 overexpression. These findings demonstrate that miR-20a-5p contributes to MPP+-induced mitochondrial disruption and cell damage, and miR-20a-5p might be a novel therapeutic target for PD.
Insights
MicroRNAs (miRNAs) show promise for Parkinson's disease (PD). This study found miR-20a-5p protects against neurotoxicity by targeting IRF9, suggesting a new therapeutic avenue for PD.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) are increasingly recognized as crucial biomarkers for Parkinson's disease (PD), influencing risk assessment, early diagnosis, and treatment strategies.
- Oxidative stress and inflammation are key pathological features implicated in the neurodegenerative processes observed in PD.
- Understanding the specific roles of individual miRNAs in modulating these cellular responses is vital for developing targeted therapies.
Purpose of the Study:
- To investigate the role of miR-20a-5p in regulating inflammation and oxidative stress within a cellular model of Parkinson's disease.
- To elucidate the underlying molecular mechanism by which miR-20a-5p exerts its effects, focusing on potential target genes.
- To evaluate the therapeutic potential of modulating miR-20a-5p levels in mitigating neurotoxicity.
Main Methods:
- Utilized the 1-methyl-4-phenyl pyridine ion (MPP+)-induced HT22 cell line as a model for Parkinson's disease neurotoxicity.
- Employed miR-20a-5p mimics and pcDNA-IRF9 to modulate gene expression, followed by MPP+ treatment.
- Assessed cell viability (CCK-8), apoptosis (flow cytometry), inflammatory and oxidative stress factors (ELISA), and protein expression (Western blot).
Main Results:
- MPP+ exposure induced mitochondrial dysfunction, inflammation, and cell damage in HT22 cells, effects which were attenuated by miR-20a-5p overexpression.
- Interferon regulatory factor 9 (IRF9) was identified as a direct target gene of miR-20a-5p.
- IRF9 overexpression exacerbated MPP+-induced neurotoxicity, mitochondrial disruption, inflammation, and apoptosis, while counteracting the protective effects of miR-20a-5p.
Conclusions:
- miR-20a-5p plays a protective role against MPP+-induced mitochondrial dysfunction and neurotoxicity in HT22 cells.
- The mechanism involves the direct targeting of IRF9, with IRF9 mediating and exacerbating the neurotoxic effects.
- miR-20a-5p represents a potential novel therapeutic target for managing Parkinson's disease.
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