| 陈雪莹,杨珂,王冬,张亮,王颖,朱慧,赵钕君,王志刚.低强度脉冲超声保护MPP+诱导的N2a细胞损伤的实验研究[J].中国医学影像技术,2020,36(2): |
| 低强度脉冲超声保护MPP+诱导的N2a细胞损伤的实验研究 |
| Protective effect of low-intensity pulsed ultrasound on N2a cells injury induced by MPP+ |
| 投稿时间：2019-07-14 修订日期：2020-02-16 |
| DOI： |
| 中文关键词: 低强度脉冲超声 帕金森病 神经调控 MPP+ N2a细胞 |
| 英文关键词:Low-intensity pulsed ultrasound Parkinson’s disease Neuromodulation MPP+ N2a cells |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 摘要点击次数: 1950 |
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| 中文摘要: |
| 目的 探讨低强度脉冲超声（LIPUS）能否减弱1-甲基-4-苯基吡啶离子（MPP+）诱导的N2a细胞毒性，对帕金森病（PD）细胞模型发挥保护作用。
方法 通过MPP+作用N2a细胞24h来建立PD细胞模型，并对N2a细胞进行超声辐照（1 MHz，50 mW/cm2，脉冲重复频率1 KHz，辐照10 min）。实验分为对照组、超声对照(LIPUS)组、MPP+组和超声处理（MPP++LIPUS）组。用CCK8法测定细胞活力，DCFH-DA染色法检测细胞内活性氧簇（ROS）的水平，JC-1染色法检测线粒体膜电位的变化，Annexin V-FITC/PI双染法检测细胞凋亡情况。实时荧光定量PCR检测瞬时受体电位M7（TRPM7）的mRNA表达水平。
结果 与对照组相比，MPP+组细胞存活率显著降低，细胞内ROS水平和线粒体膜电位上升，且细胞凋亡明显（p<0.01）；经MPP+诱导的细胞在LIPUS辐照过后，细胞存活率显著上升，细胞内ROS水平和线粒体膜电位明显下降，细胞凋亡减少（p<0.01）。MPP+组细胞中TRPM7的mRNA表达下降，而MPP++LIPUS组中TRPM7的表达显著升高（p<0.01）。
结论 LIPUS可增加MPP+诱导的N2a细胞存活率，减少线粒体膜电位损伤和ROS生成，抑制细胞凋亡，对MPP+的神经细胞毒性具有保护作用，这一作用可能与TRPM7的表达有关。 |
| 英文摘要: |
| Objective To investigate whether low-intensity pulsed ultrasound can attenuate the cytotoxicity of N2A induced by MPP+ and protect the cell model of Parkinson’s disease.
Methods N2a cells were treated with MPP+ for 24 hours to establish the PD cell model. N2A cells were irradiated by ultrasound (1 MHz, 50 mW/cm2, pulse repetition frequency 1 KHz, irradiation for 10 min). The experimental cells were divided into four groups, control group, LIPUS group, MPP+ group and MPP++LIPUS group. CCK8 assay was used to detect cell viability. The level of intracellular reactive oxygen species (ROS) was detected by DCFH-DA staining. Mitochondrial membrane potential was detected by JC-1 staining. Annexin VFITC/PI double staining method was used to detect cell apoptosis. The mRNA expression of TRPM7 was detected by real-time quantitative PCR.
Result Compared with the control group, cell viability of MPP+ group was significantly decreased, the level of intracellular ROS and mitochondrial membrane potential were increased, and cell apoptosis rate was significantly increased (p<0.01). After irradiation with LIPUS, cell viability of MPP+-induced cells was increased significantly, ROS production, mitochondrial membrane potential and cell apoptosis rate were decreased significantly (p<0.01). The mRNA expression of TRPM7 in the MPP+ group were decreased, and the expression of TRPM7 in MPP++LIPUS group were increased significantly (p<0.01).
Conclusion LIPUS can increase the survival rate of N2A cells induced by MPP+, reduce mitochondrial membrane potential damage and ROS production, inhibit apoptosis, and protect the neuronal toxicity of MPP+, which may be related to the expression of TRPM7. |
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