Ziaei Amin, Ardakani Mohammad Reza Piri, Hashemi Motahare-Sadat, Peymani Maryam, Ghaedi Kamran, Baharvand Hossein, Nasr-Esfahani Mohammad Hossein
Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Department of Cellular Biotechnology at Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran; Department of Biology, School of Sciences, University of Isfahan, Isfahan, Iran.
Neurosci Lett. 2015 Mar 17;590:138-44. doi: 10.1016/j.neulet.2015.01.083. Epub 2015 Feb 4.
Neural progenitor cells (NPCs) are feasible therapeutically model cells in regenerative medicine. However, a number of obstacles oppose their applications including insufficiency in differentiation protocols. These complications should be overwhelmed to obtain a significant clinical application. Deferoxamine (DFO), as a small molecule with a clinically high-affinity to chelate intracellular Iron, has been granted orphan drug status for treatment of traumatic spinal cord injury, while its neuroprotective function is not well understood. The aim of the present study is evaluating whether DFO could modulate neuronal differentiation process of NPCs. A varies concentrations of DFO were used to promote neuronal differentiation of mouse and human NPCs with different serum condition as an extracellular source of Iron. Several neural markers were assessed by RT-qPCR and Western analysis. Meanwhile β-catenin content was evaluated as key member of Wnt pathway. The maximal neuronal differentiation rate was observed when treating cells were treated with acute dosage of DFO (100 μM) for 6h in serum free condition. This treatment produced a significant increase in expression of neuronal markers and resulted in dramatically decrease in expression of glial markers. The protein content of β-catenin was also decreased by this treatment. Despite of chronic concentration of DFO, which reduced the size of EBs apparently due to G1/S arrest of cell cycle as known features of DFO. Application of acute courses of DFO increased neuronal differentiation rate of NPCs in serum free conditions. We concluded that suppression of Wnt/β-catenin pathway was induced through chelating of intracellular Iron due to DFO treatment. These findings help to understand therapeutic benefit of DFO as a neuroprotective agent.
神经祖细胞（NPCs）是再生医学中可行的治疗模型细胞。然而，它们的应用面临一些障碍，包括分化方案不完善。要实现显著的临床应用，必须克服这些并发症。去铁胺（DFO）作为一种对螯合细胞内铁具有临床高亲和力的小分子，已被授予孤儿药地位用于治疗创伤性脊髓损伤，但其神经保护功能尚不完全清楚。本研究的目的是评估DFO是否能调节NPCs的神经元分化过程。使用不同浓度的DFO在不同血清条件下促进小鼠和人类NPCs的神经元分化，血清作为细胞外铁源。通过RT-qPCR和Western分析评估几种神经标志物。同时，评估β-连环蛋白含量作为Wnt通路的关键成员。在无血清条件下用急性剂量的DFO（100μM）处理细胞6小时时，观察到最大的神经元分化率。这种处理使神经元标志物的表达显著增加，并导致胶质细胞标志物的表达显著降低。这种处理也降低了β-连环蛋白的蛋白质含量。尽管DFO的慢性浓度会由于细胞周期的G1/S期阻滞而明显减小EBs的大小，这是DFO已知的特征。急性疗程的DFO应用增加了无血清条件下NPCs的神经元分化率。我们得出结论，DFO处理通过螯合细胞内铁诱导了Wnt/β-连环蛋白通路的抑制。这些发现有助于理解DFO作为神经保护剂的治疗益处。
