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LINC00665/miR-132-5p Reduces Inflammation in Epileptic Cells by Targeting MAPK3
Qi-Ming Pang1, Cui Wang1, Bang-Tao Li1
1Internal Medicine-Neurology, Hainan Women and Children's Medical Center, Haikou, Hainan Province, People's Republic of China.
Abstract:
To investigate the role of miR-132-5p in the inflammatory response in epilepsy. Peripheral blood was collected from epileptic and healthy children, and the expression of LINC00665 and miR-132-5p was detected by q-PCR. Epilepsy cell models were constructed with microglia, transfected with miR-132-5p inhibitor and NC, and the expression of LINC00665 and miR-132-5p was detected by q-PCR, the expression of TNF-α, IL-1β, and IL-6 in the cell supernatant was detected by ELISA, and the protein levels of NLRP3 and MAPK3 were detected by WB. Finally, the targeting relationship between LINC00665 and miR-132-5p was verified by dual luciferase assay. The expression levels of both LINC00665 and miR-132-5p in the peripheral blood of children with epilepsy were significantly higher than those of healthy children. After transfection of epileptic cells with miR-132-5p inhibitor, the expression levels of LINC00665 and miR-132-5p were increased, and the expression levels of TNF-α, IL-1β, IL-6, NLRP3, and MAPK3 were decreased. Dual luciferase assay showed targeted binding of LINC00665 and miR-132-5p. LINC00665/miR-132-5p attenuates inflammatory responses in epileptic cells by targeting MAPK3.
Insights
The long non-coding RNA LINC00665 and microRNA miR-132-5p are elevated in pediatric epilepsy. Their interaction suppresses neuroinflammation by targeting MAPK3, offering potential therapeutic targets.
Area of Science:
- Neuroscience
- Molecular Biology
- Immunology
Background:
- Epilepsy is associated with neuroinflammation.
- MicroRNAs play crucial roles in regulating cellular processes, including inflammation.
Purpose of the Study:
- To investigate the role of miR-132-5p in the inflammatory response in epilepsy.
- To explore the relationship between LINC00665 and miR-132-5p in epilepsy.
Main Methods:
- Quantitative PCR (q-PCR) to measure gene expression.
- Enzyme-linked immunosorbent assay (ELISA) for cytokine detection.
- Western blot (WB) for protein analysis.
- Dual luciferase assay to confirm targeting interactions.
Main Results:
- LINC00665 and miR-132-5p expression were significantly higher in children with epilepsy.
- Inhibition of miR-132-5p in epilepsy cell models increased LINC00665 and miR-132-5p levels while decreasing inflammatory markers (TNF-α, IL-1β, IL-6) and NLRP3/MAPK3 proteins.
- Dual luciferase assay confirmed LINC00665 targets miR-132-5p.
Conclusions:
- LINC00665 and miR-132-5p are upregulated in pediatric epilepsy.
- The LINC00665/miR-132-5p axis attenuates inflammatory responses in epileptic cells, potentially by targeting MAPK3.
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