Department of Biophysics, National Institute of Mental Health and Neurosciences, Institute of National Importance, Hosur Road, P.B. No. 2900, Bengaluru, 560029, Karnataka, India.
Dr D.Y. Patil University (DPU), Pune, 411018, India.
Cell Mol Neurobiol. 2022 May;42(4):1167-1188. doi: 10.1007/s10571-020-01007-w. Epub 2020 Nov 18.
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 [Ca] 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 [Ca], KCl-induced [Ca], 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 Ca 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）对缺氧缺血再灌注损伤具有有益作用，但潜在机制尚不清楚。在这里，我们首先研究了 OGD 再灌注损伤对源自人胚胎干细胞（hESCs）的人 NP 细胞存活和氧化应激易感性的影响。通过测量谷胱甘肽水平、基础钙和 KCl 刺激下的细胞内钙 [Ca] 反应，以及增殖、神经胶质前体细胞标志物表达和迁移的关键参数来评估细胞失调。接下来，评估了 WJ-MSCs 恢复这些参数的影响，并评估了磷脂酰肌醇-3-激酶（PI3K）通路在激活保护作用中的作用。OGD 再灌注损伤诱导 NP 细胞死亡、ROS 生成、氧化应激易感性和谷胱甘肽水平显著增加，伴随着基础 [Ca] 升高、KCl 诱导的 [Ca] 升高、K 渗漏通道（TASK1）表达下降以及增殖、迁移潜力和神经胶质前体细胞数量下降。通过非接触方法在再灌注后 2 小时引入 WJ-MSCs，可使所有这些细胞参数在 24 小时后得到显著改善，并且 PI3K 通路在神经保护过程中发挥重要作用。WJ-MSCs 的存在增加了 OGD 再灌注 NP 中存活信号如磷酸化 Akt/Akt 和 PI3K 的表达。我们的数据首次清楚地表明，WJ-MSCs 的可溶性因子不仅可以改善 OGD 再灌注 NP 的存活、增殖、迁移和神经胶质前体细胞表达，还可以通过 PI3K 通路调节其对 KCl 刺激的细胞内 Ca 反应和 TASK1 的表达。
