RBM15 relies on m6A modification to inhibit UBE2C, alleviating hippocampal neuronal injury by limiting microglial inflammation

Abstract

Background

Microglia are strongly implicated in the development and progression of epilepsy, yet their impact on pathology remains unclear. This study aimed to explore the effects of ubiquitin-conjugating enzyme 2C (UBE2C) m6A methylation on microglial activation and neuronal injury in epilepsy.

Methods

A mouse model of pilocarpine-induced status epilepticus was constructed, and an in vitro system of HT22 hippocampal neurons was induced with Mg²⁺-free medium and cocultured with BV2 cells. The secretion of TNF-α, IL-6 and iNOS from BV2 cells was measured via qRT–PCR and ELISA. CCK-8 and flow cytometry were performed to verify cell viability and apoptosis. RNA degradation, RIP and Me-RIP assays were performed.

Results

RBM15 levels were decreased, whereas UBE2C levels were increased in the hippocampi of epileptic mice. Silencing UBE2C or overexpressing RBM15 suppressed the release of inflammatory cytokines (TNF-α and IL-6) and the M1 microglia activation marker iNOS in Mg²⁺-free BV2 cells, thereby limiting damage to hippocampal injured neurons. Mechanistically, RBM15 bound to UBE2C mRNA and decreased its stability via m6A methylation. Additionally, RBM15 inhibited the inflammatory activation of BV2 and HT22 neuron injury by inhibiting UBE2C.

Conclusion

The increase in UBE2C m6A modification induced by RBM15 upregulation inhibits hippocampal neuron damage through the suppression of microglial inflammation, providing important clues and potential targets for novel therapeutics for epilepsy.