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

IF 2.6 3区医学 Q3 NEUROSCIENCES

Molecular and Cellular Neuroscience Pub Date : 2025-03-04 DOI:10.1016/j.mcn.2025.103996

Yuehong Wan, Qin Kang, Ji Zhang, Canru Yu, Susu Fang, Dongqin Zou, Wen Chai

{"title":"RBM15 relies on m6A modification to inhibit UBE2C, alleviating hippocampal neuronal injury by limiting microglial inflammation","authors":"Yuehong Wan, Qin Kang, Ji Zhang, Canru Yu, Susu Fang, Dongqin Zou, Wen Chai","doi":"10.1016/j.mcn.2025.103996","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><div>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.</div></div><div><h3>Methods</h3><div>A mouse model of pilocarpine-induced status epilepticus was constructed, and an in vitro system of HT22 hippocampal neurons was induced with Mg<sup>2+</sup>-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.</div></div><div><h3>Results</h3><div>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 Mg<sup>2+</sup>-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.</div></div><div><h3>Conclusion</h3><div>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.</div></div>","PeriodicalId":18739,"journal":{"name":"Molecular and Cellular Neuroscience","volume":"133 ","pages":"Article 103996"},"PeriodicalIF":2.6000,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular and Cellular Neuroscience","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1044743125000065","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"NEUROSCIENCES","Score":null,"Total":0}

引用次数: 0

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.

查看原文本刊更多论文

RBM15依靠m6A修饰抑制UBE2C，通过限制小胶质细胞炎症减轻海马神经元损伤。

背景：小胶质细胞与癫痫的发生和发展密切相关，但其对病理的影响尚不清楚。本研究旨在探讨泛素偶联酶2C (UBE2C) m6A甲基化对癫痫小胶质细胞活化和神经元损伤的影响。方法：建立匹罗卡品诱导的小鼠癫痫持续状态模型，用无Mg2+培养基诱导HT22海马神经元体外系统，并与BV2细胞共培养。采用qRT-PCR和ELISA检测BV2细胞分泌TNF-α、IL-6和iNOS的水平。CCK-8和流式细胞术检测细胞活力和凋亡情况。进行RNA降解、RIP和Me-RIP测定。结果：癫痫小鼠海马区RBM15水平降低，UBE2C水平升高。沉默UBE2C或过表达RBM15可抑制无Mg2+ BV2细胞中炎症因子（TNF-α和IL-6）和M1小胶质细胞激活标志物iNOS的释放，从而限制对海马损伤神经元的损伤。机制上，RBM15结合UBE2C mRNA并通过m6A甲基化降低其稳定性。此外，RBM15通过抑制UBE2C抑制BV2的炎症激活和HT22神经元损伤。结论：RBM15上调引起的UBE2C m6A修饰增加通过抑制小胶质细胞炎症抑制海马神经元损伤，为癫痫新疗法提供重要线索和潜在靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Molecular and Cellular Neuroscience 医学-神经科学

CiteScore

5.60

自引率

0.00%

发文量

审稿时长

37 days

期刊介绍： Molecular and Cellular Neuroscience publishes original research of high significance covering all aspects of neurosciences indicated by the broadest interpretation of the journal''s title. In particular, the journal focuses on synaptic maintenance, de- and re-organization, neuron-glia communication, and de-/regenerative neurobiology. In addition, studies using animal models of disease with translational prospects and experimental approaches with backward validation of disease signatures from human patients are welcome.