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目的:研究惊厥样神经元模型中含亮氨酸重复序列和免疫球蛋白结构域的Nogo受体作用蛋白(LRR and Ig domain containing,Nogo receptor interacting protein,Lingo-1)对神经元轴突的作用。方法:(1)培养10 d的皮层神经元,经正常细胞外液处理(正常组)或无镁细胞外液处理(无镁组)后用免疫荧光技术检测神经元轴突变化。(2)以上两种不同处理后6、12、24、48、72 h 5个时间点,用q RT-PCR及Western blot检测神经元中Lingo-1表达情况。(3)设计sh RNA慢病毒抑制Lingo-1表达,分为正常组、无镁组、无镁+对照sh RNA组和无镁+Lingo-1sh RNA组4组,用免疫荧光技术检测神经元轴突长度。结果:(1)正常组神经元轴突粗壮、轴突间网络连接较致密,神经丝蛋白200(neurofilament proteins 200,NF200)只在突起表达;无镁组神经元轴突纤细、轴突间网络连接稀疏,NF200在胞体内高表达。(2)各观察时间点上,无镁组Lingo-1的表达量均高于正常组(P<0.05)。(3)在神经元轴突长度上:无镁组小于正常组(P=0.000),无镁+对照sh RNA组与无镁组无统计学差异,无镁+Lingo-1sh RNA组大于无镁组+对照sh RNA(P=0.000)。结论:无镁处理后神经元轴突损伤、Lingo-1表达升高,抑制Lingo-1表达后神经元轴突损伤减轻,说明Lingo-1可能参与神经元放电后神经元轴突损伤。
AIM: To investigate the effect of Nogo receptor interacting protein (L-nogo receptor interacting protein, Lingo-1) containing leucine repeat and immunoglobulin domains on neuronal axons in a convulsant-like neuron model . Methods: (1) Cortical neurons were cultured for 10 days. Neuronal axon changes were detected by immunofluorescence after normal extracellular fluid (normal group) or without magnesium extracellular solution (no magnesium group). (2) The expressions of Lingo-1 in neurons were detected by q RT-PCR and Western blot at 6, 12, 24, 48 and 72 h after the above two different treatments. (3) The sh RNA lentivirus was designed to inhibit Lingo-1 expression and divided into four groups: normal group, magnesium-free group, magnesium-free control sh RNA group and magnesium-free + Lingo-1sh RNA group. Sudden length. Results: (1) Neurons in the normal group had thick axons, dense interneuronal connections, and neurofilament proteins 200 (NF200) only expressed in the neurites. Magnesium-free neurons had axonal stenosis, Sparse connection, NF200 high expression in the cell body. (2) At each observation time point, the expression of Lingo-1 in the magnesium-free group was higher than that in the normal group (P <0.05). (3) In neuron axon length: no magnesium group was smaller than the normal group (P = 0.000), no magnesium + control sh RNA group and no magnesium group no statistical difference, no magnesium + Lingo-1sh RNA group than magnesium Group + control sh RNA (P = 0.000). CONCLUSION: After axon injury, the expression of Lingo-1 is increased and the axonal damage of Lingo-1 is inhibited after the treatment of magnesium-free. This indicates that Lingo-1 may be involved in neuronal axonal injury after neuronal discharge.