摘要
【目的】探讨微小非编码RNA分子miR-134在甲基苯丙胺(MA)诱导PC12神经细胞损伤中的表达变化及其对神经兴奋性的影响,为理解MA导致神经损伤的机制提供实验基础。【方法】将处于对数生长期的PC12细胞分为对照组和MA组。MA组应用800μmol/L MA处理,建立体外神经元损伤模型,观察培养神经细胞损伤及细胞凋亡情况;采用实时荧光定量PCR技术测定miR-134的表达水平变化;并构建miR-134干扰载体,抑制miR-134后分析神经诱发动作电位的变化。【结果】MA可明显诱导PC12细胞损伤,神经突起变短,细胞凋亡数目增多;荧光定量PCR结果显示miR-134表达升高;电生理学检测结果显示,干扰miR-134后诱发动作电位增多。【结论】高浓度MA诱导神经细胞损伤、诱发神经元凋亡并升高miR-134表达,沉默miR-134表达,可增加神经兴奋性。本实验为阐明MA的神经损伤机制,以及miR-134在神经元毒性损伤及神经兴奋性中的可能作用提供了实验依据。
【Objective】To investigate the expression change of miR-134 in methamphetamine（MA）-induced neuronal injury in PC12 cells and its effect on neuronal excitability and understand the pathogenesis of methamphetamine-induced neuronal injury.【Methods】PC12 cells in the logarithmic phase were divided into control group and MA group. The MA group was treated with 800μmol/L MA to establish the model of neuronal injury. The cellular injury was observed under microscope. The neuronal apoptosis was detected by Hoechst3342/PI double staining,and miR- 134 expression was measured by using real-time quantitative PCR（Real time-PCR）. Furthermore,we constructed miR-134 interference vector and observed its effect on evoked action potential.【Results】The cultured PC12 cells were damaged under the 800 μmol/LMA treatment,and neurites became shorter,the apoptotic cells were evidence. Real time-PCR showed that miR-134 expression was increased after MA treatment. Electrophysiological data showed that the evoked action potential increased after miR- 134 interference.【Conclusions】High concentration of MA can induce neuronal damage and apoptosis and also increase miR-134 expression. While silence miR-134 expression can increase neuronal excitability.Our study provides an experimental basis for elucidating the possible mechanism of MA-induced neuronal injury and the role of miR-134 in neurotoxicity and neuronal excitability.
作者
李涛
王洪杰
刘桂阳
张博
赵文博
刘天蔚
朱凯文
孙晋浩
LI Tao WANG Hong-jie LIU Gui-yang ZHANG Bo ZHAO Wen-bo LIU Tian-wei ZHU Kai-wen SUN Jin-hao(Department of Neurosurgery of Jinan No.4 People＇s Hospital, Jinan, 250031, China Department of Anatomy and Histology ＆ Embryology, Shandong University, Jinan 250012, China)
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2017年第1期36-41,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
山东省医药卫生科技发展项目(2013WS008)
山东省重点研发计划(2016GSF201054)
