Department of Biophysics, National Institute of Mental Health and Neurosciences, Institute of National Importance, Bengaluru, India.
Stem Cells Dev. 2020 Jul;29(14):929-951. doi: 10.1089/scd.2020.0018. Epub 2020 Jun 10.
In hypoxic-ischemic encephalopathy, the neural progenitors (NPs) of the developing brain fail to replenish the oligodendrocyte progenitor cells lost during hypoxic-ischemic injury (HII). In this study, we aim to examine the influence of HII on the vulnerability of human NPs derived from human embryonic stem cells with regard to cell survival and oxidative stress, followed by assessment of cellular deregulation through measuring glutathione levels, basal calcium, glutamate release, and intracellular calcium ([Ca]) response under KCl and ATP stimulation. NPs were further evaluated for their fundamental potential of self-renewal and proliferation, neural and glial progenitor pool, and migration. Oxygen-glucose deprivation (OGD) of 90 min was sublethal for NPs, yet significantly increased reactive oxygen species generation and oxidative stress susceptibility, and decreased glutathione levels, along with a rise in glutamate release, basal [Ca], and KCl and ATP-induced [Ca]. Distinct increase in gene expression for K leak channel (Twik-related acid-sensitive K channel 1 []) and purinergic receptor , and decrease of voltage-gated K channels , , and were observed. TASK-1 increase was detected by FACS too. OGD-insulted NPs showed reduced migration potential and decline in glial progenitor population. This study thus demonstrates for the first time that brief exposure of OGD does not reduce the NP population, its proliferation, and self-renewal, but can induce significant alteration in oxidative stress susceptibility, glutamate release, [Ca] response to physiological stimulus, migration, and glial progenitor pool. We thus infer that treatment strategies need to target repair of NPs of the developing brain that is affected during intrapartum asphyxia, leading to varying neurologic complications such as seizure, mental retardation, and/or cerebral palsy.
在缺氧缺血性脑病中，发育中大脑的神经祖细胞（NPs）未能补充在缺氧缺血性损伤（HII）期间丢失的少突胶质前体细胞。在这项研究中，我们旨在检查 HII 对源自人胚胎干细胞的人 NPs 的易感性的影响，具体涉及细胞存活和氧化应激，随后通过测量谷胱甘肽水平、基础钙、谷氨酸释放以及 KCl 和 ATP 刺激下的细胞内钙 ([Ca]) 反应来评估细胞失调。还进一步评估 NPs 的自我更新和增殖、神经和神经胶质祖细胞库以及迁移的基本潜能。90 分钟的氧葡萄糖剥夺（OGD）对 NPs 是亚致死的，但显着增加了活性氧生成和氧化应激易感性，并降低了谷胱甘肽水平，同时谷氨酸释放、基础 [Ca] 和 KCl 以及 ATP 诱导的 [Ca] 升高。还观察到 K 泄漏通道（Twik 相关酸敏感 K 通道 1 []) 和嘌呤能受体 的基因表达明显增加，而电压门控 K 通道 、 和 则减少。FACS 还检测到 TASK-1 的增加。OGD 损伤的 NPs 显示出迁移潜能降低和神经胶质祖细胞群减少。因此，这项研究首次证明短暂暴露于 OGD 不会减少 NP 群体、其增殖和自我更新，但会导致氧化应激易感性、谷氨酸释放、对生理刺激的 [Ca] 反应、迁移和神经胶质祖细胞库发生显着变化。因此，我们推断治疗策略需要针对在分娩期间受到影响的发育中大脑的 NPs 进行修复，这会导致各种神经并发症，如癫痫、智力迟钝和/或脑瘫。
