Division of Life Science, Advanced Research Institute of the Sciences and Humanities, Nihon University Graduate School, Tokyo, Japan.
Brain Res. 2010 Oct 28;1358:20-9. doi: 10.1016/j.brainres.2010.08.048. Epub 2010 Aug 22.
We hypothesized that one of the mechanisms underlying the protection of brain injury by therapeutic hypothermia is associated with preservation of neural stem cells. We investigated effects of moderate low temperature and the contribution of a cold-inducible molecule for the stemness of neural stem cells. The MEB5 mouse neural stem cell line was cultured in the presence or absence of EGF, and apoptosis, mRNA expression, and immunocytochemistry of the differentiation markers nestin and GFAP were evaluated at 37 or 32°C. We investigated the contribution of the cold-inducible RNA binding protein (CIRP) on apoptosis and differentiation of MEB5 cells at 32°C. EGF deprivation increased the number of apoptotic cells, decreased expression of nestin, and increased expression of GFAP. The moderate low temperature prevented apoptosis and decreases in expression of GFAP in MEB5 by EGF deprivation. The moderate low temperature significantly increased expression of CIRP. siRNA against CIRP significantly increased the apoptotic cell population of MEB5 cells via EGF deprivation at 32°C. These findings suggest that moderate low temperature preserved stemness of neural stem cells and prevented cell apoptosis via the stimulation of CIRP, and one of the mechanisms of rescue of brain injury by the moderate hypothermia is associated with preservation of neural stem cells.
我们假设治疗性低温对脑损伤的保护机制之一与神经干细胞的保护有关。我们研究了中低温的影响以及冷诱导分子对神经干细胞干性的贡献。在存在或不存在 EGF 的情况下培养 MEB5 小鼠神经干细胞系，并在 37°C 或 32°C 下评估细胞凋亡、mRNA 表达和分化标志物巢蛋白和 GFAP 的免疫细胞化学。我们研究了冷诱导 RNA 结合蛋白 (CIRP) 在 32°C 下对 MEB5 细胞凋亡和分化的贡献。EGF 剥夺增加了凋亡细胞的数量，降低了巢蛋白的表达，增加了 GFAP 的表达。中低温通过 EGF 剥夺防止 MEB5 细胞的凋亡和 GFAP 表达的降低。中低温显著增加了 CIRP 的表达。CIRP 的 siRNA 显著增加了 EGF 剥夺时 MEB5 细胞的凋亡细胞群。这些发现表明，中低温通过刺激 CIRP 保存了神经干细胞的干性，并防止了细胞凋亡，而中度低温对脑损伤的抢救机制之一与神经干细胞的保护有关。
