Chen Shu, Luo Ming, Zhao Yuming, Zhang Yimin, He Mingliang, Cai Wangqing, Liu Anmin
Cell Physiol Biochem. 2015;36(2):531-41. doi: 10.1159/000430118.
Neurite outgrowth is one of the important therapeutic strategies for neuronal plasticity and regeneration in neural disorders. Fasudil is a clinical medication that is used to treat subarachnoid haemorrhage (SAH) and that is beneficial for many animal models of central nervous system (CNS) diseases. In this study, we hypothesised that fasudil administration would promote neurite outgrowth in neural stem cells (NSCs).
Changes in cell morphology were imaged under a light microscope, and neurite-bearing cells were counted. Cell viability and the necrosis rate were determined by MTT and LDH assays, respectively. Additionally, western blot and immunofluorescence analyses were performed to detect protein expression levels.
We found that fasudil promoted neurite outgrowth in C17.2 cells in a time- and dose-dependent manner. The neurite-bearing C17.2 cells were differentiated by detecting the changes in neural and astrocytic markers after fasudil treatment through down-regulating Notch signalling. Previously, fasudil was reported to induce autophagy, which plays an important role in neural differentiation. However, both rapamycin, an autophagy inducer, and 3-methyl-adenine (3-MA), an autophagy inhibitor, had no effects on the fasudil-induced neurite outgrowth, suggesting that autophagy may be not involved in this process.
In summary, fasudil could stimulate neurite outgrowth and differentiation in C17.2 cells by modulating Notch signalling but not autophagy.
神经突生长是神经疾病中神经元可塑性和再生的重要治疗策略之一。法舒地尔是一种临床药物，用于治疗蛛网膜下腔出血（SAH），对许多中枢神经系统（CNS）疾病的动物模型有益。在本研究中，我们假设给予法舒地尔会促进神经干细胞（NSCs）的神经突生长。
在光学显微镜下对细胞形态变化进行成像，并对有神经突的细胞进行计数。分别通过MTT和LDH测定法测定细胞活力和坏死率。此外，进行蛋白质印迹和免疫荧光分析以检测蛋白质表达水平。
我们发现法舒地尔以时间和剂量依赖性方式促进C17.2细胞的神经突生长。通过下调Notch信号通路，检测法舒地尔处理后神经和星形胶质细胞标志物的变化，使有神经突的C17.2细胞分化。以前有报道说法舒地尔可诱导自噬，自噬在神经分化中起重要作用。然而，自噬诱导剂雷帕霉素和自噬抑制剂3-甲基腺嘌呤（3-MA）对法舒地尔诱导的神经突生长均无影响，这表明自噬可能不参与此过程。
总之，法舒地尔可通过调节Notch信号通路而非自噬来刺激C17.2细胞的神经突生长和分化。
