Identification of plasma exosomal microRNAs and bioinformatics analysis of the microRNA-messenger RNA regulatory pathways in mice with status epilepticus.
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Abstract
Status epilepticus (SE) is a serious neurological emergency that brings significant risks to health and life. microRNAs (miRNAs) and their targets show involvement in the pathophysiology of SE. We identified plasma exosomal miRNAs and analyzed the miRNA-messenger RNA (mRNA) regulatory pathways in SE mice. Mice were subjected to SE induction by kainic acid injection, and plasma exosome (Exo) extraction. Exo morphology, particle size distribution, and Exo-positive marker proteins were evaluated. Differentially-expressed miRNAs in Exos of SE mice were analyzed and verified by sequencing and RT-qPCR. Functional enrichment analysis on target genes and protein-protein interaction (PPI) network were performed. Hippocampal neuron cells HT-22 were cultured in vitro, and the targeted binding association between Exos-derived miR-205-5p and target genes was invalidated. There were 64 differentially-expressed miRNAs in plasma Exos of SE mice from healthy mice (32 up-regulated, 32 down-regulated). Among the top 10 differentially-expressed miRNAs, 5 were up-regulated, and 5 were down-regulated. The PPI network of collective target genes was developed, including 11 edges and 9 nodes. The genes related to nerve injury were phosphatase and tensin homolog (Pten), glycogen synthase kinase 3 beta (Gsk3b), and leucine-rich repeat kinase 2 (Lrrk2). SE mouse plasma Exos targeted Gsk3b, Lrrk2 and Pten in neuronal cells and reduced cell viability. Plasma exosomal miRNAs of SE mice were differentially expressed, and their target genes participated in the regulation of multiple pathways, mainly related to nervous system development. miR-205-5p could target Gsk3b, Lrrk2 and Pten, and suppress neuronal viability.
Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00708-8.
期刊介绍：
The scope of the Journal includes:
1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products.
2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools.
3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research.
4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy.
5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.
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