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Published on: May 17, 2024
Role of rno-miR-124-3p in regulating MCT1 expression in rat brain after permanent focal cerebral ischemia
Shi-Ye Xu1, Xu-Li Jiang1, Qian Liu1
1Department of Anatomy, Chongqing Medical University 1 Yixueyuan Road, Yuzhong District, China.
Abstract:
This study aimed to assess the role of microRNAs (miRNAs) in regulating monocarboxylate transporter-1 (MCT1) expression in rat brain after permanent focal cerebral ischemia to identify a new target for early treatment of cerebral ischemia. Focal cerebral ischemia was induced by permanent middle cerebral artery occlusion (pMCAO) in rats. Morphology and protein expression levels of MCT1 were assessed by immunofluorescence and Western blotting. Using bioinformatics and double luciferase reporter assays, rno-miR-124-3p was selected as a direct target for rat MCT1. Expression of rno-miR-124-3p after pMCAO was detected. Then, rats were treated with rno-miR-124-3p agomir via lateral ventricle injection, and after 6 h or 24 h ischemia, rno-miR-124-3p expression and gene and protein expression of MCT-1 were detected by qRT-PCR and Western blotting. Brain infarction was identified by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. Results showed that pMCAO induced brain infarction and increased the expression of MCT1. The levels of rno-miR-124-3p after pMCAO were in contrast to those of MCT1 protein in ischemic region, while declined after 3, 6 and 12 h of pMCAO in ischemic penumbra. After administration of rno-miR-124-3p agomir, MCT1 mRNA and protein levels were increased after 6 h of pMCAO, while decreased after 24 h of pMCAO. Meanwhile, rno-miR-124-3p levels increased after both times. TTC staining showed treatment with rno-miR-124-3p agomir reduced brain infarction. The role of rno-miR-124-3p in regulating MCT1 was as a positive regulator after 6 h of pMCAO, while a negative regulator after 24 h of pMCAO, however, both activities had protective effects against cerebral ischemia.
Insights
MicroRNAs (miRNAs) regulate monocarboxylate transporter-1 (MCT1) in rat brain stroke. Targeting rno-miR-124-3p shows potential for early cerebral ischemia treatment by reducing brain infarction.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cerebral ischemia, often caused by middle cerebral artery occlusion (MCAO), leads to brain infarction.
- Monocarboxylate transporter-1 (MCT1) plays a role in brain energy metabolism and is implicated in ischemic conditions.
- MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and are involved in cellular responses to injury.
Purpose of the Study:
- To investigate the role of specific miRNAs in regulating MCT1 expression following permanent focal cerebral ischemia in rats.
- To identify potential miRNA-based therapeutic targets for early intervention in cerebral ischemia.
Main Methods:
- Permanent focal cerebral ischemia was induced in rats using permanent middle cerebral artery occlusion (pMCAO).
- MCT1 expression was analyzed using immunofluorescence and Western blotting.
- Bioinformatics and luciferase reporter assays identified rno-miR-124-3p as a direct target of rat MCT1.
- Rats were treated with rno-miR-124-3p agomir, and outcomes including miRNA/MCT1 expression and infarct volume (TTC staining) were assessed.
Main Results:
- pMCAO induced brain infarction and increased MCT1 expression, while rno-miR-124-3p levels decreased in the ischemic penumbra.
- Administration of rno-miR-124-3p agomir modulated MCT1 mRNA and protein levels at different time points post-ischemia (6h vs. 24h).
- Treatment with rno-miR-124-3p agomir significantly reduced brain infarction volume.
- rno-miR-124-3p acted as a positive regulator of MCT1 at 6h and a negative regulator at 24h, with both effects demonstrating protective benefits.
Conclusions:
- Rno-miR-124-3p directly targets and regulates MCT1 expression in the context of cerebral ischemia.
- The dual role of rno-miR-124-3p as a regulator of MCT1, acting as both a positive and negative regulator at different time points, offers a novel therapeutic strategy.
- Targeting rno-miR-124-3p presents a promising avenue for early treatment of cerebral ischemia.
