Guo Minfang, Zhang Huiyu, Yu Jingwen, Zhang Peijun, Wang Yuyin, Wei Wenyue, Song Lijuan, Chai Zhi, Yu Jiezhong, Ma Cungen
Institute of Brain Science/Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases, Shanxi Datong University, Datong 037009, China.
Institute of Brain Science/Shanxi Key Laboratory of Inflammatory Neurodegenerative Diseases, Shanxi Datong University, Datong 037009; The Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Research Center for Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2022 Jul;38(7):625-631.
Objective To investigate the effect of Fasudil on HO-induced apoptosis and synaptic plasticity in human neuroblastoma SY5Y cells and its mechanism. Methods The cells were divided into three groups: PBS control group, HO model group (250 μmol/L HO treatment) and Fasudil intervention group (250 μmol/L HO combined with 15 μg/mL Fasudil treatment). MTT assay was applied to detect cell activity and TUNEL was performed to detect cell apoptosis respectively. Immunofluorescence cytochemical staining was used to determine the expression of neurite outgrowth inhibitor A (NogoA), Nogo receptor (NgR) and synaptophysin (Syn). Western blotting was then conducted to detect the expression of NogoA, NgR, p75 neurotrophin receptor (p75NTR), leucine-rich repeat Ig domain-containing Nogo-interacting protein 1 (LINGO-1), Syn and postsynaptic density protein-95 (PSD-95). Results Compared with the PBS group, the HO group showed decreased cell viability and increased apoptosis rate while Fasudil treatment significantly increased the cell viability and reduced the apoptosis rate. Compared with the HO model group, Fasudil intervention increased expressions of Syn and PSD-95. Compared with the PBS group, the expression of NogoA and its receptor complex NgR/p75NTR/LINGO-1 grew significantly in the HO group, suggesting Fasudil treatment could inhibit the expression of NogoA and its receptor complex NgR/p75NTR/LINGO-1. Conclusion Fasudil may inhibit the activation of the NogoA/NgR signaling pathway, therefore reducing the apoptosis induced by HO in SH-SY5Y cells and enhancing the plasticity of the synapses.
目的 探讨法舒地尔对人神经母细胞瘤SY5Y细胞中血红素加氧酶（HO）诱导的细胞凋亡及突触可塑性的影响及其机制。方法 将细胞分为三组：PBS对照组、HO模型组（250 μmol/L HO处理）和法舒地尔干预组（250 μmol/L HO联合15 μg/mL法舒地尔处理）。分别采用MTT法检测细胞活性，TUNEL法检测细胞凋亡。采用免疫荧光细胞化学染色法检测神经突生长抑制因子A（NogoA）、Nogo受体（NgR）和突触素（Syn）的表达。然后进行蛋白质免疫印迹法检测NogoA、NgR、p75神经营养因子受体（p75NTR）、富含亮氨酸重复Ig结构域的Nogo相互作用蛋白1（LINGO-1）、Syn和突触后致密蛋白95（PSD-95）的表达。结果 与PBS组相比，HO组细胞活力降低，凋亡率升高，而法舒地尔处理显著提高了细胞活力，降低了凋亡率。与HO模型组相比，法舒地尔干预使Syn和PSD-95的表达增加。与PBS组相比，HO组中NogoA及其受体复合物NgR/p75NTR/LINGO-1的表达显著增加，提示法舒地尔处理可抑制NogoA及其受体复合物NgR/p75NTR/LINGO-1的表达。结论 法舒地尔可能抑制NogoA/NgR信号通路的激活，从而减少HO诱导的SH-SY5Y细胞凋亡，增强突触可塑性。
