离体癫痫持续状态模型中抑制miR-134对树突棘的影响  

The role of miR134 in remodeling of dendritic spines in status epilepticus model in vitro

作　　者：王小木[1] 李德帅[1] 姜曌 万群 王津存[1] 王卫蝉 夏峰[1] Wang Xiaomu;Li Deshuai;Jiang Zhao;Wan Qun;Wang Jincun;Wang weichan;Xia Feng(Departmeng of Neurology,the Xijing Hospital,Fourth Military Medical University,Xian 710032,China)

机构地区：[1]第四军医大学西京医院神经内科,西安710032 [2]天津解放军254医院神经内科

出　　处：《脑与神经疾病杂志》2018年第9期559-563,共5页Journal of Brain and Nervous Diseases

基　　金：陕西省自然基金项目(2014K12-01-01)

摘　　要：目的探讨抑制miR-134对原代培养神经元树突棘的影响。方法 (1)构建大鼠miR-134抑制慢病毒载体并验证;(2)培养海马神经元,建立无镁离体癫痫持续状态SE模型,利用离子扫描显微镜研究miR-134抑制对癫痫持续状态SE模型中神经元树突棘密度的影响。结果 (1)将一段362bp包含2个miR-134竞争结合位点的DNA片段经XhoI和EcoRI酶切后插入质粒pCDH-CMV-MCS-EG(R)FP-MCS中E(R)GFP序列的下游,该片段转录产物与miR-134互补形成稳定的复合物,从而阻止了miR-134对靶mRNA的降解或翻译抑制,得到大鼠miR-134抑制慢病毒载体;(2)同对照病毒组相比,miR-134抑制慢病毒预处理组经无镁细胞外液处理后(离体癫痫持续状态模型)神经元树突棘密度减少(P<0.05)。结论 MiR-134可能在树突棘重塑过程中扮演重要角色。Objective To investigate the role of miR134 in remodeling of dendritic spines in status epilepticus(SE)model in vitro.Method ①Rat miR-134 inhibited lentiviral vectors were constructed and verified for subsequent studies.②In vitro model of SE,the influence of knock down of miR-134 by miR-134 inhibited lentiviral vectors on the neuronal dendritic spines was observed.Results①A synthesized 362 bp long DNA fragment which contain two miR-134 competitive binding sites was inserted into downstream of EG(R)FP sequences in pCDH-CMV-MCS-EG(R)FP-MCS vector to get rat miR-134 inhibitor lentiviral vector.The transcript of DNA fragments can specifically bind to miR-134 to form a stable complex,which can inhibit the degradation and translation suppression of miR-134 to its targets.MiR-134 inhibitor recombinant lentivirus containing GFP(or RFP)was constructed;②Compared with the control virus group,the neuronal dendritic spines density in the miR-134 inhibitor recombinant lentivirus group was decreased in the SE model that induced by low-Mg2+extracellular fluid(P<0.05).Conclusion MiR-134 may play an important role in remodeling of dendritic spines.

关 键 词：癫痫持续状态 miR-134 慢病毒 树突棘 

分 类 号：R742.1[医药卫生—神经病学与精神病学]