Targeted inhibition of microglial C5aR1 by PMX205 mitigates post-ischemic stroke neuroinflammation and promotes functional recovery

IF 3.7 3区医学 Q2 NEUROSCIENCES

Brain Research Bulletin Pub Date : 2026-04-06 DOI:10.1016/j.brainresbull.2026.111873

Jie Cao , Saisai Tian , Zilong Deng , Jin Xu , Yangyang Liu , Xuan Shi , Lei Bai , Xiang Li Jr , Haiying Li

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引用次数: 0

Abstract

Background

Ischemic stroke constitutes the leading cause of death and disability worldwide. Post-stroke neuroinflammation, a major driver of secondary neurodegeneration, has emerged as a priority therapeutic target. Microglia serve as critical initiators of this neuroinflammatory cascade in ischemic stroke. This study employed single-cell RNA sequencing and functional experiments to identify key regulatory factors in microglia following distinct ischemic stroke subtypes, with the goal of translating these findings into therapeutic targets for clinical application.

Methods

To identify key regulators in ischemic stroke, we mined and analyzed public single-cell RNA sequencing datasets. Two etiologically distinct stroke models were subsequently established: permanent focal ischemia via distal middle cerebral artery occlusion (dMCAO) and transient ischemia-reperfusion injury using middle cerebral artery occlusion/reperfusion (MCAO/R). C5aR1 spatiotemporal expression was quantified through immunofluorescence (cellular localization) and quantitative immunoblotting (temporal dynamics), followed by validation of PMX205-C5aR1 binding affinity via rigid-receptor molecular docking. Therapeutic assessment included acute-phase measurements (3d post-stroke): pro-inflammatory cytokines (ELISA), cerebral infarction volume (TTC staining), and co-quantification of neuronal apoptosis/viability (TUNEL/Nissl); alongside chronic functional recovery tracking (14d): motor coordination (rotarod), sensorimotor integration (adhesive removal test), anxiety-like behavior (open field exploration), and spatial working memory (Y-maze spontaneous alternation).

Results

Bioinformatics analysis identified significant upregulation of C5aR1 in activated microglia following ischemic stroke. This finding was corroborated by spatially resolved immunofluorescence and quantitative immunoblotting; molecular docking confirmed stable PMX205-C5aR1 binding via specific hydrophobic interactions, and subsequent therapeutic intervention with PMX205 profoundly suppressed neuroinflammation (IL-1β/IL-6/TNF-α), reduced cerebral infarction, attenuated neuronal apoptosis, and reversed long-term neurological deficits.

Conclusion

Targeted inhibition of C5aR1 by PMX205 represents a therapeutically viable strategy to attenuate neuroinflammatory cascades and improve long-term functional recovery after ischemic stroke.

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PMX205靶向抑制小胶质细胞C5aR1减轻缺血性脑卒中后神经炎症并促进功能恢复。

背景：缺血性中风是世界范围内死亡和残疾的主要原因。脑卒中后神经炎症是继发性神经变性的主要驱动因素，已成为优先治疗的目标。小胶质细胞是缺血性卒中中神经炎症级联反应的关键启动因子。本研究采用单细胞RNA测序和功能实验来鉴定不同缺血性卒中亚型小胶质细胞的关键调控因子，目的是将这些发现转化为临床应用的治疗靶点。方法：为了确定缺血性卒中的关键调控因子，我们挖掘和分析了公开的单细胞RNA测序数据集。随后建立了两种病因不同的脑卒中模型：通过大脑中动脉远端闭塞（dMCAO）建立永久性局灶性缺血模型和通过大脑中动脉闭塞/再灌注（MCAO/R）建立短暂性缺血-再灌注损伤模型。通过免疫荧光（细胞定位）和定量免疫印迹（时间动力学）定量C5aR1时空表达，然后通过刚性受体分子对接验证PMX205-C5aR1结合亲和力。治疗评估包括急性期测量（卒中后3d）：促炎细胞因子（ELISA）、脑梗死体积（TTC染色）和神经元凋亡/活力联合量化（TUNEL/Nissl）；同时进行慢性功能恢复跟踪（14d）：运动协调（rottarod）、感觉运动整合（黏附去除测试）、类焦虑行为（开放领域探索）和空间工作记忆（y迷宫自发交替）。结果：生物信息学分析发现，缺血性卒中后激活的小胶质细胞中C5aR1显著上调。空间分辨免疫荧光和定量免疫印迹证实了这一发现；分子对接证实PMX205- c5ar1通过特异性的恐水相互作用稳定结合，随后PMX205治疗干预可深度抑制神经炎症（IL-1β/IL-6/TNF-α），减少脑梗死，减轻神经元凋亡，逆转长期神经功能缺损。结论：PMX205靶向抑制C5aR1是一种治疗上可行的策略，可以减轻缺血性脑卒中后的神经炎症级联反应，改善长期功能恢复。

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

Brain Research Bulletin 医学-神经科学

CiteScore

6.90

自引率

2.60%

发文量

253

审稿时长

67 days

期刊介绍： The Brain Research Bulletin (BRB) aims to publish novel work that advances our knowledge of molecular and cellular mechanisms that underlie neural network properties associated with behavior, cognition and other brain functions during neurodevelopment and in the adult. Although clinical research is out of the Journal''s scope, the BRB also aims to publish translation research that provides insight into biological mechanisms and processes associated with neurodegeneration mechanisms, neurological diseases and neuropsychiatric disorders. The Journal is especially interested in research using novel methodologies, such as optogenetics, multielectrode array recordings and life imaging in wild-type and genetically-modified animal models, with the goal to advance our understanding of how neurons, glia and networks function in vivo.