1 Department of Neurology, Hanyang University College of Medicine, Seoul, Korea.
2 Department of Translational Medicine, Hanyang University Graduate School of Biomedical Science & Engineering, Seoul, Korea.
Stem Cells Dev. 2019 Jun 15;28(12):769-780. doi: 10.1089/scd.2018.0138. Epub 2019 Apr 22.
Cerebral infarction is one of the major causes of severe morbidity and mortality, and thus, research has focused on developing treatment options for this condition. Zinc (Zn) is an essential element in the central nervous system and has several neuroprotective effects in the brain. In this study, we examined the neuroprotective effects of Zn on neural stem cells (NSCs) exposed to hypoxia. After treatment with several concentrations of Zn, the viability of NSCs under hypoxic conditions was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Trypan blue staining, and a lactate dehydrogenase assay. To evaluate the effect of Zn on the proliferation of NSCs, bromodeoxyuridine/5-bromo-2'-deoxyuridine (BrdU) labeling and colony formation assays were performed. Apoptosis was also examined in NSCs exposed to hypoxia with and without Zn treatment. In addition, a western blot analysis was performed to evaluate the effect of Zn on intracellular signaling proteins. NSC viability and proliferation were decreased under hypoxic conditions, but treatment with sublethal doses of Zn restored viability and proliferation. Sublethal doses of Zn reduced apoptosis caused by hypoxia, increased the expression levels of proteins related to the phosphatidylinositol-3 kinase (PI3K) pathway, and decreased the expression levels of proteins associated with neuronal cell death. These findings confirm that in vivo, sublethal doses of Zn protect NSCs against hypoxia through the activation of the PI3K pathway. Thus, Zn could be employed as a therapeutic option to protect NSCs in ischemic stroke.
脑梗死是严重发病率和死亡率的主要原因之一,因此,研究集中在开发这种疾病的治疗选择上。锌(Zn)是中枢神经系统中的必需元素,在大脑中有多种神经保护作用。在这项研究中,我们研究了 Zn 对缺氧暴露的神经干细胞(NSC)的神经保护作用。在用几种浓度的 Zn 处理后,通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)测定法、台盼蓝染色和乳酸脱氢酶测定法来测量缺氧条件下 NSC 的活力。为了评估 Zn 对 NSCs 增殖的影响,进行了溴脱氧尿苷/5-溴-2'-脱氧尿苷(BrdU)标记和集落形成测定。还检查了缺氧条件下有无 Zn 处理对 NSCs 凋亡的影响。此外,进行了 Western blot 分析以评估 Zn 对细胞内信号蛋白的影响。在缺氧条件下,NSC 的活力和增殖减少,但亚致死剂量的 Zn 处理恢复了活力和增殖。亚致死剂量的 Zn 减少了缺氧引起的凋亡,增加了与磷脂酰肌醇-3 激酶(PI3K)途径相关的蛋白的表达水平,并降低了与神经元细胞死亡相关的蛋白的表达水平。这些发现证实,在体内,亚致死剂量的 Zn 通过激活 PI3K 途径来保护 NSCs 免受缺氧的影响。因此,Zn 可以作为治疗选择来保护缺血性中风中的 NSCs。