小鼠皮层神经元机械损伤后miR-124的动态变化及意义  

Dynamic change and significance of miR-124 in vitro after mechanical damage of cortical neurons in mice

作　　者：苏鑫洪 叶玉勤[1] 杨永祥[1] 贺晓生[1] 

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2017年第3期229-233,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81471264)

摘　　要：目的观察体外培养小鼠皮层神经元机械损伤后微小核糖核酸-124(miR-124)的表达变化,初步探讨miR-124对小鼠创伤性脑损伤后神经轴突再生与修复可能的影响。方法孕15~18 d C57BL/6种孕鼠胚胎脑皮质层神经元体外培养7 d,以10μL移液器塑料滴头在培养皿内划割,造成机械性损伤,伤后不同时间点(1 h,6 h,12 h,24 h,72 h,144 h)分别采用实时定量聚合酶链法(RTPCR)检测miR-124和蛋白质印记法(Western Blot)检测神经纤毛蛋白(Nrp-1)、微管相关蛋白(Tau)、生长相关蛋白43(Gap-43)的表达水平。然后用miR-124模拟物和抑制剂分别对体外培养的皮层神经元进行干预,观察miR-124表达量的改变对实验的影响。结果神经元机械损伤后miR-124和Nrp-1、Tau、Gap-43的表达均显著升高(P<0.05),且存在一定的正相关性。用miR-124模拟物和抑制剂分别显著升高和降低miR-124的表达后,Nrp-1、Tau、Gap-43表达显著下降,其中抑制剂组下降较模拟物组明显(P<0.05)。结论创伤区miR-124的适度高表达可能与轴突再生有密切联系,这为本课题组今后尝试梯度调控miR-124以调节神经轴突再生与修复提供了实验依据。Objective The changes of microRNA-124 （miR-124） in vitro after mechanical damage of cortical neurons, and the influence of miR-124 on axon regeneration and repair after traumatic brain injury in mice were explored.Methods Primary cortical neurons were obtained from fetal C57BL/6 mice and cultivated for 7 d. Petri dishes were manually scratched with a 10 μL plastic stylet needle following a 9 ×9 square grim. Cells were collected at 1 h, 6 h, 12 h, 24 h, 72 h and 144 h after injury for experiments. The real-time polymerase chain reaction （RT- PCR） and Western Blot were used to test the relative expressions of miR-124 and Neuropilin-1 （Nrp-1）, microtubule- associated protein tau （Tau）, growth associated protein 43 （Gap-43）, respectively. Then miR-124 mimic and inhibitor were used to intervene the cortex neuron in vitro to observe how the change of miR-124 expression quantity affected the experiment, Results MiR-124, Nrp-1, Tau and Gap-43 were significantly increased after mechanical damage of cortical neurons （P 〈0.05） and there was a positive correlation between them. The miR-124 mimic and inhibitor were significantly increased and decreased the expression of miR-124 respectively and then caused a significant reduction in the expressions of Nrp-1, Tau, Gap-43 and the decrease in inhibitor group was more significant than that of mimic group （P 〈0.05）. Condusion Moderate high expression of miR-124 may be closely related to the axon regeneration. The result provides the experimental basis for regulating the expression of miR-124 gradually to regulate neural axon regeneration and restoration.

关 键 词：机械损伤 微小核糖核酸-124 轴突再生修复 

分 类 号：R651[医药卫生—外科学]