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MicroRNA-599 regulates the development of Parkinson's disease through mediating LRRK2 expression
1Department of Rehabilitation Medicine, Shanxi Provincial People's Hospital, Taiyuan, China. liyujun-wuqin@163.com.
Objective:
This study investigates whether microRNA-599 can inhibit the progression of Parkinson's disease (PD) by regulating the LRRK2 expression. We aim to search for a new therapeutic target for PD.
Materials And Methods:
A mouse model of PD was first established. A relative amount of TH+ neurons in the mouse brain was quantified by immunohistochemistry. The expression levels of microRNA-599 and LRRK2 in mouse brain tissues were determined by the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Cell model of PD was constructed by MPP+ treatment in SH-SY5Y cells. The expression levels of microRNA-599 and LRRK2 in MPP+-induced SH-SY5Y cells were examined as well. We verified the binding condition between microRNA-599 and LRRK2 through dual-luciferase reporter gene assay. The viability and apoptosis in MPP+-induced SH-SY5Y cells overexpressing microRNA-599 were determined by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively.
Results:
Compared with normal mice, TH+ neurons were fewer in the brain tissue of PD mice. MicroRNA-599 expression was lower, while LRRK2 expression was higher in brain tissues of PD mice relative to controls. Meanwhile, in vitro expression of microRNA-599 was downregulated and LRRK2 expression was upregulated in MPP+-induced SH-SY5Y cells. Dual-luciferase reporter gene assay verified the binding condition between microRNA-599 and LRRK2. The microRNA-599 overexpression downregulated the LRRK2 expression in SH-SY5Y cells, and conversely, the microRNA-599 knockdown upregulated the LRRK2 expression. Of note, the microRNA-599 overexpression protected MPP+-induced viability decrease and apoptosis acceleration in SH-SY5Y cells.
Conclusions:
MicroRNA-599 is lowly expressed in both in vivo and in vitro PD model. MicroRNA-599 inhibits the development of PD through regulating the LRRK2 expression.
Insights
MicroRNA-599, a key molecule, is found at low levels in Parkinson's disease (PD) models. Increasing microRNA-599 can inhibit PD progression by regulating LRRK2 expression, offering a potential therapeutic strategy.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Parkinson's disease (PD) is a progressive neurodegenerative disorder.
- The precise molecular mechanisms underlying PD pathogenesis remain incompletely understood.
- Identifying novel therapeutic targets is crucial for effective PD treatment.
Purpose of the Study:
- To investigate the role of microRNA-599 in Parkinson's disease progression.
- To determine if microRNA-599 regulates Leucine-Rich Repeat Kinase 2 (LRRK2) expression in PD.
- To explore microRNA-599 as a potential therapeutic target for PD.
Main Methods:
- Establishment of a mouse model of Parkinson's disease.
- Quantification of dopaminergic neurons (TH+ neurons) using immunohistochemistry.
- Measurement of microRNA-599 and LRRK2 expression via qRT-PCR and Western blot.
- Construction of an in vitro PD cell model (MPP+ treatment in SH-SY5Y cells).
- Verification of microRNA-599 and LRRK2 interaction using dual-luciferase reporter gene assay.
- Assessment of cell viability (CCK-8 assay) and apoptosis (flow cytometry) in response to microRNA-599 manipulation.
Main Results:
- PD mouse models exhibited reduced TH+ neurons, decreased microRNA-599, and increased LRRK2 expression compared to controls.
- In vitro PD models showed downregulated microRNA-599 and upregulated LRRK2.
- Dual-luciferase assay confirmed direct binding between microRNA-599 and LRRK2.
- Overexpression of microRNA-599 reduced LRRK2 levels and protected SH-SY5Y cells from MPP+-induced viability loss and apoptosis.
Conclusions:
- MicroRNA-599 expression is significantly downregulated in both in vivo and in vitro models of Parkinson's disease.
- MicroRNA-599 exerts an inhibitory effect on PD development.
- This inhibition is mediated through the regulation of Leucine-Rich Repeat Kinase 2 (LRRK2) expression, highlighting a novel therapeutic avenue.