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| Anosmin-1蛋白通过成纤维生长因子1型受体激活ERK1/2信号通路增强嗅鞘细胞的迁移 |
| Anosmin-1 protein enhances the migration of olfactory ensheathing cells by fibroblast growth factor 1 type receptor activating ERK1/2 signaling pathway |
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| DOI：10.12089/jca.2021.05.017 |
| 中文关键词: Anosmin-1蛋白 嗅鞘细胞 迁移 FGF 1型受体 ERK1/2信号通路 神经型钙粘连蛋白 |
| 英文关键词: Anosmin-1 protein Olfactory ensheathing cell Migration FGF 1 type receptor ERK1/2 signaling pathway N-cadherin |
| 基金项目:国家自然科学基金（81571193） |
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| 中文摘要: |
目的 探究Anosmin-1蛋白对嗅鞘细胞迁移的影响及对成纤维生长因子（FGF）1型受体和ERK1/2信号通路的影响。
方法 选择SPF级成年雌性SD大鼠40只，7~10周龄，体重250~300 g。提取大鼠嗅球的嗅鞘细胞并纯化，纯化后细胞培养7 d后，采用随机数字表法将所有细胞平均分为四组：对照组（C组），FGF 1型受体激动剂（FGF2）组（F组），Anosmin-1 蛋白组（A组）和Anosmin-1 蛋白+FGF 1型受体抑制剂（Su5402）组（S组）。C组为空白对照组，培养液中不加任何药物处理；F组为阳性对照组，于培养液中加入FGF2 25 ng/ml；A组于培养液中加入Anosmin-1蛋白2 nmol/L；S组于培养液中加入Anosmin-1蛋白2 nmol/L和Su5402 30 μmol/L。四组细胞处理24 h后，采用Transwell法检测嗅鞘细胞迁移能力，Western blot法检测嗅鞘细胞磷酸化ERK1/2（p-ERK1/2）蛋白、磷酸化AKT（p-AKT）蛋白和神经型钙粘连蛋白（N-cadherin）含量，免疫荧光染色法检测N-cadherin荧光强度。
结果 纯化后的嗅鞘细胞占总细胞的85.6%。与C组比较，F组和A组迁移至下室的嗅鞘细胞明显增多（P<0.05），p-ERK1/2蛋白、N-cadherin含量明显升高（P<0.05）。与A组比较，S组迁移至下室的嗅鞘细胞明显减少（P<0.05），p-ERK1/2蛋白、N-cadherin含量明显降低（P<0.05）。S组和C组间、F组和A组间迁移至下室的嗅鞘细胞数量、p-ERK1/2蛋白、N-cadherin含量差异无统计学意义。四组p-AKT蛋白含量差异无统计学意义。
结论 Anosmin-1蛋白通过FGF 1型受体激活ERK1/2信号通路，从而上调N-cadherin，增强嗅鞘细胞的迁移能力。 |
| 英文摘要: |
Objective To investigate the effect and the molecular mechanism of Anosmin-1 protein on the migration of olfactory ensheathing cells and the effect of fibroblast growth factor (FGF) 1 type receptor and ERK1/2 signaling pathway.
Methods Forty healthy female Sprague-Dawley rats were chosen, aged 7-10 weeks, weighing 250-300 g. The olfactory ensheathing cells from olfactory bulb of rats were extracted, purified and cultured for 7 days. After culturing, all cells were equally divided into four groups according random number table method: control group (group C), FGF 1 type receptor agonist (FGF2) treated group (group F), Anosmin-1 protein treated group (group A) and Anosmin-1 protein add to FGF 1 type receptor inhibitor (Su5402) treated group (group S). Group C was blank control group, group F was positive control treated with FGF2 25 ng/ml, group A was treated with Anosmin-1 protein 2 nmol/L, group S was treated with Anosmin-1 protein 2 nmol/L added to Su5402 30 μmol/L. The migration ability of olfactory ensheathing cells was detected by Transwell migration assay. The expression of phosphorylated ERK1/2 (p-ERK1/2), phosphorylated AKT (p-AKT), and N-cadherin downsteaming of FGF signaling pathway was detected by Western blot after treatment with Anosmin-1, Su5402 and FGF2. Immunofluorescence was used to reveal the fluorescence intensity of N-cadherin after treatments of Anosmin-1, Su5402 and FGF2.
Results The purified olfactory ensheathing cells accounted for 85.6% of the total cells. Compared with group C, the migratory amount of olfactory ensheathing cells of groups F and Awere significantly increased (P < 0.05), the expressions of p-ERK1/2 protein and N-cadherin were significantly increased (P < 0.05), and the fluorescence intensity of N-cadherin were significantly increased (P < 0.05). Compared with group A, the migratory amount of olfactory ensheathing cells of group S was significantly decreased (P < 0.05), the expressions of p-ERK1/2 protein and N-cadherin were significantly decreased (P < 0.05), and the fluorescence intensity of N-cadherin was significantly decreased (P < 0.05). There was no significantly differences in migratory capacity, expressions of p-ERK1/2 protein and N-cadherin, and the fluorescence intensity of N-cadherin between groups C and S, and groups F and A. There was no significant diferences in expression of p-AKT protein in each groups.
Conclusion Anosmin-1 protein enhances the migratory ability of olfactory ensheathing cells through FGF 1 type receptor activating ERK1/2 signaling pathway and the upregulation of N-cadherin expression． |
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