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【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 of miR-134, a small non-coding RNA molecule, in the injury induced by methamphetamine (PC) in PC12 cells and its effect on neuronal excitability, and provide the experimental basis for understanding the mechanism of MA-induced neuronal injury. 【Method】 PC12 cells in logarithmic growth phase were divided into control group and MA group. MA group was treated with 800μmol / L MA to establish a model of neuronal injury in vitro, to observe the damage of cultured nerve cells and the apoptosis of cells; to detect the expression level of miR-134 by real-time fluorescence quantitative PCR; to construct the miR-134 interference vector, After inhibition of miR-134, neuro-induced action potentials were analyzed. 【Result】 MA could obviously induce the injury of PC12 cells, shorten the number of neurites and increase the number of apoptotic cells. Fluorescence quantitative PCR results showed that the expression of miR-134 was increased. Electrophysiology results showed that the action potential was increased after interference with miR-134. 【Conclusion】 High concentrations of MA can induce neuronal damage, induce neuronal apoptosis and increase the expression of miR-134, silence the expression of miR-134, and increase the excitability of nerve. This experiment provides an experimental basis for elucidating the mechanism of nerve damage in MA and the possible role of miR-134 in neuronal damage and neuronal excitability.