Phosphorylation of cAMP-Activated Exchange Protein-1 Participates in Neuroprotection and Ferroptosis Regulation Following Intracerebral Hemorrhage in Rats

IF 4.8 1区医学 Q1 NEUROSCIENCES

CNS Neuroscience & Therapeutics Pub Date : 2025-04-09 DOI:10.1111/cns.70373

Guannan Jiang, Jialei Zhou, Yan Zhuang, Siyuan Yang, Gang Chen, Wanchun You, Xiang Li

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Abstract

Background

Intracerebral hemorrhage (ICH) is a severe condition characterized by elevated mortality and disability rates. The cAMP-activated exchange protein-1 (EPAC-1) is implicated in various cytoprotective mechanisms; however, its specific role in ICH remains unclear.

Methods

A rat model of ICH was established by injecting autologous blood, while the in vitro primary neuronal model was stimulated using oxyhemoglobin (OxyHb). The construction of EPAC-1 overexpression wild-type (WT) and phosphorylated mutant plasmids (S108A or S108E), as well as lentiviruses, was performed for in vitro and in vivo studies.

Results

The cAMP signaling pathway was found to be significantly enriched following ICH by high-throughput sequencing analysis. Our findings showed that while EPAC-1 protein levels remained relatively unchanged after ICH, RabGEF activity was conspicuously upregulated. This was accompanied by a marked decrease in EPAC-1 phosphorylation levels. Mutations that activate EPAC-1 phosphorylation led to significant improvements in neuronal survival and behavioral outcomes after ICH. Bioinformatics analysis revealed that ferroptosis was significantly enriched after ICH and showed a positive correlation with EPAC-1 serine phosphorylation. EPAC-1 phosphorylation activating mutations inhibit neuronal ferroptosis, whereas inactivating mutations exacerbate it.

Conclusion

The phosphorylation of EPAC-1 is essential for maintaining neuronal survival, which may be related to ferroptosis inhibition after ICH.

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cAMP 激活的交换蛋白-1 磷酸化参与大鼠脑出血后的神经保护和铁蛋白沉积调节

脑出血（ICH）是一种严重的疾病，其特点是死亡率和致残率升高。camp活化交换蛋白-1 （EPAC-1）参与多种细胞保护机制；然而，其在非ICH中的具体作用尚不清楚。方法采用自体血注射法建立脑出血大鼠模型，并用氧合血红蛋白（OxyHb）刺激体外原代神经元模型。构建EPAC-1过表达野生型（WT）和磷酸化突变质粒（S108A或S108E）以及慢病毒，进行体外和体内研究。结果经高通量测序分析，发现ICH后cAMP信号通路显著富集。我们的研究结果表明，ICH后EPAC-1蛋白水平保持相对不变，而RabGEF活性明显上调。这伴随着EPAC-1磷酸化水平的显著下降。激活EPAC-1磷酸化的突变导致脑出血后神经元存活和行为结果的显著改善。生物信息学分析显示，ICH后铁下垂显著富集，且与EPAC-1丝氨酸磷酸化呈正相关。EPAC-1磷酸化激活突变抑制神经元铁下垂，而失活突变加剧。结论EPAC-1的磷酸化对维持脑出血后神经元存活至关重要，可能与脑出血后铁下垂抑制有关。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

CNS Neuroscience & Therapeutics 医学-神经科学

CiteScore

7.30

自引率

12.70%

发文量

240

审稿时长

2 months

期刊介绍： CNS Neuroscience & Therapeutics provides a medium for rapid publication of original clinical, experimental, and translational research papers, timely reviews and reports of novel findings of therapeutic relevance to the central nervous system, as well as papers related to clinical pharmacology, drug development and novel methodologies for drug evaluation. The journal focuses on neurological and psychiatric diseases such as stroke, Parkinson’s disease, Alzheimer’s disease, depression, schizophrenia, epilepsy, and drug abuse.