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目的探讨转铁蛋白受体1(TfR1)对小鼠海马神经元生长与分化的影响。方法取胚胎18.5 d小鼠原代海马神经元培养,培养7 d后使用TfR1 shRNA p GFP-V-RS干扰质粒转染小鼠海马原代神经元使TfR1基因沉默,采用绿色荧光蛋白(GFP)分析及Western blotting技术检测TfR1 shRNA的转染效率;4’6-二脒基-2-苯基吲哚(DAPI)及TUNEL法检测干扰后海马神经细胞凋亡情况,免疫荧光技术检测凋亡后神经细胞骨架的变化及神经元突起生长的形态特征。结果 TfR1 shRNA质粒能有效使神经细胞内TfR1基因沉默;TfR1基因沉默后,原代海马神经细胞凋亡率增加,细胞骨架部分崩解,且神经元树突突起长度明显增长(P<0.05)。结论转铁蛋白受体1可通过调控小鼠海马神经元树突的生长来影响海马神经元的分化。
Objective To investigate the effects of transferrin receptor 1 (TfR1) on the growth and differentiation of hippocampal neurons in mice. Methods Primary cultured hippocampal neurons of embryonic day 18.5 d were cultured. Primary cultured hippocampal neurons were transfected with TfR1 shRNA p GFP-V-RS interference plasmids to silence TfR1 gene after 7 days of culture. Green fluorescent protein (GFP) The transfection efficiency of TfR1 shRNA was analyzed and analyzed by Western blotting. The apoptotic rate of hippocampal neurons was detected by 4’6-diamidino-2-phenylindole (DAPI) and TUNEL assay, and the apoptosis was detected by immunofluorescence Changes of neuronal cytoskeleton and morphological features of neurite outgrowth. Results The TfR1 shRNA plasmid effectively silenced TfR1 gene in neurons. After TfR1 gene silencing, the apoptosis rate of primary hippocampal neurons increased, the cytoskeleton partially disintegrated, and the length of dendrites of neurons increased significantly (P <0.05). Conclusion Transferrin receptor 1 can affect the differentiation of hippocampal neurons by regulating the dendritic growth of hippocampal neurons in mice.