Irreversible brain injury and neurological dysfunction induced by cardiac arrest (CA) have long been a clinical challenge due to lack of effective therapeutic interventions to reverse neuronal loss and prevent secondary reperfusion injury. The neuronal regenerative potential of neural stem cells (NSCs) provides a possible solution to this clinical deficit. We investigated the neuronal recovery potential of human neural stem cells (hNSCs) via intracerebroventricular (ICV) xenotransplantation after CA in rats and the effects of transplanted NSCs on the proliferation and migration of endogenous NSCs. Outcome measures included neurological functional recovery measured by neurological deficit score (NDS), electrophysiologic analysis of EEG, and assessment of proliferation and migration at the cellular level and the Wnt/β-catenin pathway at the molecular level. Neurological functional assessment based on aggregate neurological deficit score (NDS) showed better recovery of function after hNSCs therapy (P < 0.05). Tracking of stem cells’ proliferation with Ki67 antibody suggested that the NSCs group had more prominent proliferation compared to control group (number of Ki67+ cells, Control VS. NSC: 89.0 ± 31.6 VS. 352.7 ± 97.3, P < 0.05). In addition, cell migration tracked by Dcx antibody showed more Dcx + cells migrated to the far distance zone from SVZ in the treatment group (P < 0.05). Further immunofluorescence staining confirmed that the expression of the Wnt signaling pathway protein (β-catenin) was upregulated in the NSC group (P < 0.05). ICV delivery of hNSCs promotes endogenous NSC proliferation and migration and ultimately enhances neuronal survival and neurological functional recovery. Wnt/β-catenin pathway may be involved in the initiation and maintenance of this enhancement.

Graphical abstract

由于缺乏有效的治疗干预措施来逆转神经元丢失和预防继发性再灌注损伤，由心脏骤停 (CA) 引起的不可逆脑损伤和神经功能障碍长期以来一直是临床挑战。神经干细胞 (NSC) 的神经元再生潜力为这种临床缺陷提供了可能的解决方案。我们通过大鼠 CA 后脑室内 (ICV) 异种移植研究了人神经干细胞 (hNSCs) 的神经元恢复潜力，以及移植的 NSCs 对内源性 NSCs 增殖和迁移的影响。结果测量包括通过神经功能缺损评分 (NDS) 测量的神经功能恢复、EEG 的电生理分析、评估细胞水平的增殖和迁移以及分子水平的 Wnt/β-catenin 通路。基于综合神经功能缺损评分 (NDS) 的神经功能评估显示 hNSCs 治疗后功能恢复更好。P  < 0.05)。用 Ki67 抗体跟踪干细胞增殖表明，与对照组相比，NSCs 组具有更显着的增殖（Ki67+ 细胞数，Control VS. NSC：89.0 ± 31.6 VS. 352.7 ± 97.3，P  < 0.05）。此外，Dcx抗体追踪的细胞迁移显示，治疗组有更多的Dcx + 细胞迁移到离SVZ较远的区域（P < 0.05)。进一步免疫荧光染色证实，NSC组Wnt信号通路蛋白（β-catenin）的表达上调（P < 0.05）。hNSCs 的 ICV 递送促进内源性 NSC 增殖和迁移，并最终增强神经元存活和神经功能恢复。Wnt/β-catenin 通路可能参与了这种增强的启动和维持。

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