Increasing evidence suggests that mesenchymal stem cells(MSCs) have beneficial effects in hypoxic ischemic reperfusion injury, but the underlying mechanisms are unclear. Here, we first examined the effect of OGD reperfusion injury on the vulnerability of human NPs derived from human embryonic stem cells (hESCs) with regard to cell survival and oxidative stress. Cellular deregulation was assessed by measuring glutathione levels, basal calcium and intracellular calcium [Ca²⁺]_i response under KCl stimulation, as well as the key parameters of proliferation, glial progenitor marker expression and migration. Next, the influence of WJ-MSCs in recovering these parameters was evaluated, and the role of Phosphatidyl-inositol-3-Kinase(PI3K) pathway in actuating the protective effect was assessed. OGD reperfusion injury induced significant increases in cell death, ROS generation, oxidative stress susceptibility and decreased glutathione levels in NPs, accompanied by rises in basal [Ca²⁺]_i, KCl-induced [Ca²⁺]_i, expression of K⁺ leak channel(TASK1), and declines in proliferation, migration potential and glial progenitor population. The introduction of WJ-MSCs(after 2 h of reperfusion) through a non-contact method brought about significant improvement in all these cellular parameters as observed after 24hrs, and the PI3K pathway played an important role in the neuroprotection process. Presence of WJ-MSCs increased the expression of survival signals like phosphorylated Akt/Akt and PI3K in the OGD-reperfused NPs. Our data clearly demonstrate for the first time that soluble factors from WJ-MSCs can not only ameliorate survival, proliferation, migration and glial progenitor expression of OGD-reperfused NPs, but also regulate their intracellular Ca²⁺ response to KCl stimulation and expression of TASK1 through the PI3K pathway.

越来越多的证据表明间充质干细胞（MSCs）在缺氧缺血再灌注损伤中具有有益作用，但其潜在机制尚不清楚。在这里，我们首先检查了 OGD 再灌注损伤对源自人类胚胎干细胞 (hESCs) 的人类 NPs 在细胞存活和氧化应激方面的脆弱性的影响。通过测量谷胱甘肽水平、基础钙和细胞内钙 [Ca ²⁺ ] _{i评估细胞失调}KCl 刺激下的反应，以及增殖、胶质祖细胞标志物表达和迁移的关键参数。接下来，评估了 WJ-MSCs 在恢复这些参数中的影响，并评估了磷脂酰肌醇 3-激酶 (PI3K) 途径在激活保护作用中的作用。OGD再灌注损伤导致NPs细胞死亡、活性氧生成、氧化应激敏感性显着增加，谷胱甘肽水平降低，同时基础[Ca ²⁺ ] _i、KCl诱导的[Ca ²⁺ ] _i、K ^{+表达升高}泄漏通道（TASK1），增殖、迁移潜力和胶质祖细胞数量下降。通过非接触方法引入 WJ-MSCs（再灌注 2 小时后）带来了 24 小时后观察到的所有这些细胞参数的显着改善，并且 PI3K 通路在神经保护过程中发挥了重要作用。WJ-MSCs 的存在增加了 OGD 再灌注 NPs 中生存信号的表达，如磷酸化 Akt/Akt 和 PI3K。我们的数据首次清楚地表明，来自 WJ-MSCs 的可溶性因子不仅可以改善 OGD 再灌注 NPs 的存活、增殖、迁移和神经胶质祖细胞的表达，还可以调节它们对 KCl 刺激的细胞内 Ca ²⁺反应和 TASK1 的表达通过 PI3K 途径。