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Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy
Published on: August 5, 2014
Molecular expression and functional analysis of genes in children with temporal lobe epilepsy
Xiaojuan Wu1, Yajie Wang1, Zhenrong Sun2
1Department of Pediatrics, Beijing Tiantan Hospital, Capital Medical University, Beijing, 100050, China.
Insights
MicroRNA-135a-5p is elevated in pediatric temporal lobe epilepsy, promoting apoptosis and reducing cell survival by inhibiting caspase activity and apoptosis inhibitor 1. This finding offers potential therapeutic targets for this common childhood epilepsy.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Temporal lobe epilepsy (TLE) is the most prevalent epilepsy syndrome.
- Antiseizure medications are often ineffective in treating pediatric TLE.
- MicroRNAs (miRNAs) play a role in TLE development, suggesting therapeutic potential.
Purpose of the Study:
- To investigate the expression and function of miR-135a-5p in pediatric TLE.
- To determine the relationship between miR-135a-5p, caspase activity, and apoptosis inhibitor 1 in TLE.
- To elucidate the role of miR-135a-5p in hippocampal cell apoptosis and survival in TLE.
Main Methods:
- Analysis of miR-135a-5p expression in hippocampal tissues from pediatric TLE patients and controls.
- In vitro culture of primary hippocampal neurons from rats using a magnesium-free medium to model TLE.
- Assessment of apoptosis and cell proliferation using flow cytometry and MTT assays, respectively.
- Evaluation of miR-135a-5p inhibitor transfection effects on cellular processes.
Main Results:
- miR-135a-5p levels were significantly increased in pediatric TLE patients and the in vitro epilepsy model.
- Overexpression of miR-135a-5p led to downregulation of caspase activity and apoptosis inhibitor 1.
- miR-135a-5p promoted apoptosis and reduced cell survival in hippocampal cells under TLE conditions.
Conclusions:
- miR-135a-5p acts as a pro-apoptotic factor in pediatric TLE by suppressing caspase activity and apoptosis inhibitor 1.
- Elevated miR-135a-5p contributes to reduced cell survival in pediatric TLE.
- Targeting miR-135a-5p may represent a novel therapeutic strategy for pediatric TLE.
Abstract:
Temporal lobe epilepsy is the most common form of epilepsy. However, for this type of condition, antiseizure medication is not effective for children. As miRNAs are involved in the development of temporal lobe epilepsy in children, they may provide potential therapeutic approaches for treatment. The primary aim of this study was to explore the expression and function of miR-135a-5p in children with temporal lobe epilepsy. Hippocampal slices from either normal (control) children or children with temporal lobe epilepsy were used to detect the expression of miR-135a-5p and its target gene caspase activity and apoptosis inhibitor 1. To further explore the role of miR-135a-5p in the development of temporal lobe epilepsy in children, primary hippocampal neurons from newborn rats were cultured in vitro in a magnesium-free medium to mimic the temporal lobe epilepsy condition in children. The effect of transfection of miR-135a-5p inhibitor into cells was also assessed. Apoptosis and proliferation of hippocampus cells was respectively assessed by flow cytometry or 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The level of miR-135a-5p was significantly increased in both children with temporal lobe epilepsy and the epileptiform discharge model that employed newborn rat hippocampal neurons; whereas, the expression of caspase activity and apoptosis inhibitor 1 was downregulated by overexpression of miR-135a-5p. Moreover, miR-135a-5p mediated the pro-apoptotic effect of temporal lobe epilepsy via repressing caspase activity and apoptosis inhibitor 1 expression. Additionally, miR-135a-5p reduced cell survival in the temporal lobe epilepsy condition. Overexpression of miR-135a-5p induced cell apoptosis through inhibition of caspase activity and apoptosis inhibitor 1 expression and suppressed cell survival in children with temporal lobe epilepsy.
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