甲基转移酶样3介导的小胶质细胞焦亡在败血症相关脑病中的机制。

IF 1.7 4区医学 Q4 NEUROSCIENCES

Neuroreport Pub Date : 2025-11-05 Epub Date: 2025-09-05 DOI:10.1097/WNR.0000000000002215

Dandan Chi, Feng Li, Zhimin Wang

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引用次数: 0

摘要

目的：脓毒症相关脑病（SAE）是脓毒症常见且严重的神经系统并发症。本研究旨在探讨甲基转移酶样3 （METTL3）在SAE诱导的小胶质细胞焦亡中的作用机制，并寻找SAE治疗的新靶点。方法：采用脂多糖（LPS）处理的BV-2细胞建立SAE细胞模型。检测白细胞介素-1β、白细胞介素-18、cleaved caspase-1、gasdermin D (GSDMD)-N、nod样受体蛋白3 （NLRP3）、转化生长因子受体3 （TGFBR3）、METTL3的表达。在lps处理的BV-2细胞中，METTL3被沉默，以验证METTL3在小胶质细胞焦亡中的作用。测定总n6 -甲基腺苷（m6A）含量。采用甲基化RNA免疫沉淀- qpcr分析初级miRNA (pri-miR)-101-3p与DGCR8的结合以及pri-miR-101-3p的m6A水平。逆转录定量PCR检测miR-101-3p和miR-101-3p的表达。通过数据库预测miR-101-3p的下游靶点，验证miR-101-3p与TGFBR3的结合关系。通过救援实验验证METTL3/miR-101-3p/TGFBR3轴在小胶质细胞焦亡中的作用。结果：LPS处理降低细胞活力，促进白细胞介素-1β、白细胞介素-18、METTL3、cleaved caspase-1、GSDMD-N和NLRP3的表达。沉默METTL3抑制小胶质细胞焦亡。机制上，METTL3通过m6A修饰促进pri-miR-101-3p与DGCR8结合，增加成熟miR-101-3p的表达。miR-101-3p靶向TGFBR3并抑制TGFBR3的表达。miR-101-3p过表达或TGFBR3下调部分逆转了沉默METTL3对lps诱导的小胶质细胞焦亡的抑制作用。结论：METTL3在SAE中表达上调，通过m6A修饰增强miR-101-3p表达，抑制TGFBR3表达，最终导致SAE小胶质细胞焦亡。

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Mechanisms of methyltransferase-like 3-mediated microglial pyroptosis in sepsis-associated encephalopathy.

Objective: Sepsis-associated encephalopathy (SAE) is a common and serious neurological complication of sepsis. This study aimed to investigate the mechanism of methyltransferase-like 3 (METTL3) in SAE-induced microglial pyroptosis and to identify new therapeutic targets for SAE treatment.

Methods: A SAE cell model was established using lipopolysaccharide (LPS)-treated BV-2 cells. The expression of interleukin-1β, interleukin-18, cleaved caspase-1, gasdermin D (GSDMD)-N, NOD-like receptor protein 3 (NLRP3), transforming growth factor beta receptor 3 (TGFBR3), and METTL3 was detected by. METTL3 was silenced in LPS-treated BV-2 cells to validate the role of METTL3 in microglial pyroptosis. Total N6-methyladenosine (m6A) content was measured. The binding of primary miRNA (pri-miR)-101-3p to DGCR8 and the m6A level of pri-miR-101-3p were analyzed by methylated RNA immunoprecipitation-qPCR. The expression of pri-miR-101-3p and miR-101-3p was measured by reverse transcription quantitative PCR. The downstream targets of miR-101-3p were predicted by databases, and the binding relationship between miR-101-3p and TGFBR3 was verified. Rescue experiments were performed to verify the role of METTL3/miR-101-3p/TGFBR3 axis in microglial pyroptosis.

Results: LPS treatment decreased cell viability and promoted interleukin-1β, interleukin-18, METTL3, cleaved caspase-1, GSDMD-N, and NLRP3. Silencing METTL3 inhibited microglial pyroptosis. Mechanistically, METTL3 promoted the binding of pri-miR-101-3p to DGCR8 through m6A modification and increased mature miR-101-3p expression. miR-101-3p targeted TGFBR3 and inhibited TGFBR3 expression. miR-101-3p overexpression or TGFBR3 downregulation partially reversed the inhibitory effect of silencing METTL3 on LPS-induced microglial pyroptosis.

Conclusion: METTL3 is upregulated in SAE, enhances miR-101-3p expression through m6A modification, and inhibits TGFBR3 expression, finally leading to microglial pyroptosis in SAE.

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来源期刊

Neuroreport 医学-神经科学

CiteScore

3.20

自引率

0.00%

发文量

150

审稿时长

1 months

期刊介绍： NeuroReport is a channel for rapid communication of new findings in neuroscience. It is a forum for the publication of short but complete reports of important studies that require very fast publication. Papers are accepted on the basis of the novelty of their finding, on their significance for neuroscience and on a clear need for rapid publication. Preliminary communications are not suitable for the Journal. Submitted articles undergo a preliminary review by the editor. Some articles may be returned to authors without further consideration. Those being considered for publication will undergo further assessment and peer-review by the editors and those invited to do so from a reviewer pool. The core interest of the Journal is on studies that cast light on how the brain (and the whole of the nervous system) works. We aim to give authors a decision on their submission within 2-5 weeks, and all accepted articles appear in the next issue to press.