PDF(Pubmed)
Abstract：
OBJECTIVE: Postoperative cognitive dysfunction (POCD) is prevalent among the elderly, characterized primarily by cognitive decline after surgery. This study aims to explore how extracellular vesicles (EVs) derived from BV2 microglial cells, with and without the C-C chemokine receptor type 5 (CCR5), affect neuroinflammation, neuronal integrity, and cognitive function in a POCD mouse model.
METHODS: We collected EVs from LPS-stimulated BV2 cells expressing CCR5 (EVsM1) and from BV2 cells with CCR5 knockdown (EVsM1-CCR5). These were administered to POCD-induced mice. Protein interactions between CCR5, G-protein-coupled receptors (GPCRs), and Ras were analyzed using structure-based docking and co-immunoprecipitation (Co-IP). We assessed the phosphorylation of p38 and Erk, the expression of synaptic proteins PSD95 and MAP2, and conducted Morris Water Maze tests to evaluate cognitive function.
RESULTS: Structure-based docking and Co-IP confirmed interactions between CCR5, GPR, and Ras, suggesting a CCR5-GPCRs-Ras-MAPK pathway involvement in neuroinflammation. EVsM1 heightened neuroinflammation, reduced synaptic integrity, and impaired cognitive function in POCD mice. In contrast, EVsM1-CCR5 reduced neuroinflammatory markers, preserved synaptic proteins, enhanced dendritic spine structure, and improved cognitive outcomes.
CONCLUSIONS: EVsM1 induced neuroinflammation via the CCR5-GPCRs-Ras-MAPK pathway, with EVsM1-CCR5 showing protective effects on POCD progression, suggesting a new therapeutic strategy for POCD management via targeted modification of microglial EVs.
METHODS: We collected EVs from LPS-stimulated BV2 cells expressing CCR5 (EVsM1) and from BV2 cells with CCR5 knockdown (EVsM1-CCR5). These were administered to POCD-induced mice. Protein interactions between CCR5, G-protein-coupled receptors (GPCRs), and Ras were analyzed using structure-based docking and co-immunoprecipitation (Co-IP). We assessed the phosphorylation of p38 and Erk, the expression of synaptic proteins PSD95 and MAP2, and conducted Morris Water Maze tests to evaluate cognitive function.
RESULTS: Structure-based docking and Co-IP confirmed interactions between CCR5, GPR, and Ras, suggesting a CCR5-GPCRs-Ras-MAPK pathway involvement in neuroinflammation. EVsM1 heightened neuroinflammation, reduced synaptic integrity, and impaired cognitive function in POCD mice. In contrast, EVsM1-CCR5 reduced neuroinflammatory markers, preserved synaptic proteins, enhanced dendritic spine structure, and improved cognitive outcomes.
CONCLUSIONS: EVsM1 induced neuroinflammation via the CCR5-GPCRs-Ras-MAPK pathway, with EVsM1-CCR5 showing protective effects on POCD progression, suggesting a new therapeutic strategy for POCD management via targeted modification of microglial EVs.
摘要：
目的：术后认知功能障碍（POCD）在老年人中普遍存在，主要表现为手术后认知能力下降。本研究旨在探讨BV2小胶质细胞来源的细胞外囊泡(EV),有和没有C-C趋化因子受体5型（CCR5），影响神经炎症，神经元的完整性，和POCD小鼠模型中的认知功能。
方法：我们从表达CCR5的LPS刺激的BV2细胞（EVsM1）和CCR5敲低的BV2细胞（EVsM1-CCR5）收集EV。将这些施用于POCD诱导的小鼠。CCR5，G蛋白偶联受体（GPCRs）之间的蛋白质相互作用，和Ras使用基于结构的对接和免疫共沉淀(Co-IP)进行分析。我们评估了p38和Erk的磷酸化，突触蛋白PSD95和MAP2的表达,并进行Morris水迷宫测试以评估认知功能。
结果：基于结构的对接和Co-IP确认了CCR5，GPR，还有Ras,提示CCR5-GPCRs-Ras-MAPK通路参与神经炎症。EVsM1加剧了神经炎症，突触完整性降低，和POCD小鼠的认知功能受损。相比之下,EVsM1-CCR5降低了神经炎症标志物，保存的突触蛋白，增强的树突脊柱结构，和改善认知结果。
结论：EVsM1通过CCR5-GPCRs-Ras-MAPK通路诱导神经炎症，EVsM1-CCR5对POCD进展具有保护作用，提出了一种通过靶向修饰小胶质细胞电动汽车来管理POCD的新治疗策略。
方法：我们从表达CCR5的LPS刺激的BV2细胞（EVsM1）和CCR5敲低的BV2细胞（EVsM1-CCR5）收集EV。将这些施用于POCD诱导的小鼠。CCR5，G蛋白偶联受体（GPCRs）之间的蛋白质相互作用，和Ras使用基于结构的对接和免疫共沉淀(Co-IP)进行分析。我们评估了p38和Erk的磷酸化，突触蛋白PSD95和MAP2的表达,并进行Morris水迷宫测试以评估认知功能。
结果：基于结构的对接和Co-IP确认了CCR5，GPR，还有Ras,提示CCR5-GPCRs-Ras-MAPK通路参与神经炎症。EVsM1加剧了神经炎症，突触完整性降低，和POCD小鼠的认知功能受损。相比之下,EVsM1-CCR5降低了神经炎症标志物，保存的突触蛋白，增强的树突脊柱结构，和改善认知结果。
结论：EVsM1通过CCR5-GPCRs-Ras-MAPK通路诱导神经炎症，EVsM1-CCR5对POCD进展具有保护作用，提出了一种通过靶向修饰小胶质细胞电动汽车来管理POCD的新治疗策略。
