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miR-128-3p inhibits the inflammation by targeting MAPK6 in penicillin-induced astrocytes
Yuejiu Pang1, Dingzhen Luo1, Shuhua Wang2
1Department of Senile Neurology.
Objective:
Epilepsy causes physical and mental damage to patients. As well known, microRNAs (miRNAs) provide therapeutic target potentials for patients with epilepsy. miR-128-3p was previously reported to be downregulated in temporal lobe epilepsy (TLE) patients, however, its detailed function in epilepsy is unknown.
Methods:
Astrocytes function in epilepsy, penicillin-induced astrocytes can be used as a model for seizures in vitro. Currently, the expression levels of mitogen-activated protein kinase 6 (MAPK6), interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) were determined by western blot and reverse transcription-quantitative PCR analyses (RT-qPCR). The expression level of miR-128-3p was evaluated by RT-qPCR. TargetScan 7.1 and dual luciferase reporter assay were used for prediction and verification of interaction between miR-128-3p and MAPK6 3' untranslated region (UTR). Cell viability was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay.
Results:
We found that penicillin-induced decrease in cell viability, and increase of TNF-α/IL-1β in primary astrocytes. There were lower miR-128-3p and higher MAPK6 in penicillin-treated primary astrocytes. miR-128-3p overexpression rescued penicillin-induced reduction of cell viability, and upregulation of TNF-α/IL-1β, which was partially abolished by MAPK6 overexpression.
Conclusion:
Altogether, miR-128-3p attenuates penicillin-induced cell injury and inflammation in astrocytes by targeting MAPK6, thus providing a protective role in epilepsy.
Insights
MicroRNA-128-3p protects against epilepsy-related astrocyte damage by targeting MAPK6. This finding offers a potential therapeutic strategy for epilepsy by modulating inflammation and cell injury.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Epilepsy poses significant physical and mental health challenges.
- MicroRNAs (miRNAs) are emerging as potential therapeutic targets for epilepsy.
- miR-128-3p is downregulated in temporal lobe epilepsy (TLE), but its specific role remains unclear.
Purpose of the Study:
- To investigate the function of miR-128-3p in epilepsy.
- To explore the regulatory relationship between miR-128-3p and MAPK6 in astrocytes.
- To assess the therapeutic potential of miR-128-3p in an epilepsy model.
Main Methods:
- Primary astrocytes were treated with penicillin to model seizures in vitro.
- Expression levels of miR-128-3p, MAPK6, IL-1β, and TNF-α were quantified using RT-qPCR and Western blot.
- Cell viability was assessed using MTT assays, and miR-128-3p/MAPK6 interactions were validated via dual-luciferase reporter assays.
Main Results:
- Penicillin-induced seizures decreased astrocyte viability and increased TNF-α/IL-1β levels.
- miR-128-3p expression was reduced, while MAPK6 expression was elevated in penicillin-treated astrocytes.
- Overexpression of miR-128-3p restored cell viability and reduced inflammation, an effect partially reversed by MAPK6 overexpression.
Conclusions:
- miR-128-3p mitigates penicillin-induced astrocyte injury and inflammation by targeting MAPK6.
- miR-128-3p demonstrates a protective role in epilepsy.
- Targeting miR-128-3p may offer a novel therapeutic approach for epilepsy treatment.
