M2 Microglia-Derived Exosomes Protect Against Glutamate-Induced HT22 Cell Injury via Exosomal miR-124-3p

• DOI: 10.1007/s12035-024-04075-x
• Publication Date: 2024-03-04T04:10:11Z
• Authors: Lan Zhu; Limei Ma; Xin Du; Yuhao Jiang; Jiake Gao; Zihao Fan; Hengheng Zheng; Jianjun Zhu; Gaofeng Zhang
• Keywords: Exosome; Viability assay

As one of the most serious complications sepsis, sepsis-associated encephalopathy has not been effectively treated or prevented. Exosomes, as a new therapeutic method, play protective role in neurodegenerative diseases, stroke and traumatic brain injury recent years. The purpose this study was to investigate exosomes glutamate (Glu)-induced neuronal injury, explore its mechanism, providing ideas for treatment encephalopathy. neuron damage model induced by Glu established, metabolomics analyzed identified. BV2 cells were differentiate into M1 M2 subtypes. After from both M1-BV2 M2-BV2 collected, exosome morphological identification performed transmission electron microscopy exosome-specific markers also detected. These then cocultured with HT22 cells. CCK-8 method LDH kit used detect cell viability toxicity. Cell apoptosis, mitochondrial membrane potential ROS content respectively detected flow cytometry, JC-1 assay DCFH-DA assay. MiR-124-3p expression level qRT-PCR Western blot. Bioinformatics analysis luciferase reporter predicted verified relationship between miR-124-3p ROCK1 ROCK2. Through metabolomics, 81 different metabolites found, including fructose, GABA, 2, 4-diaminobutyric acid, etc. enrichment differential showed that they mainly enriched glutathione metabolism, glycine serine urea cycle. microglia-derived could reduce decrease accumulation ROS, restore anti-oxidative stress ability Glu. It found effect mimic on neurons comparable M2-EXOs. Additionally, M2-EXOs might carry target ROCK2 neurons, affecting ROCK/PTEN/AKT/mTOR signaling pathway, reducing Glu-induced apoptosis. may protect against transferring cells, ROCK being gene miR-124-3p.