丹皮酚通过抑制线粒体氧化应激防止1-甲基-4-苯基吡啶离子损伤SH-SY5Y细胞

Paeonol Protects SH-SY5Y cells from MPP^+-induced Damage by Inhibiting Mitochondrial Oxidative Stress

目的探究丹皮酚(Paeonol,Pae)防止1-甲基-4-苯基吡啶离子(1-methyl-4-phenyl pyridine,MPP+)损伤神经细胞的机制。方法采用MPP+作用于人神经母细胞瘤细胞株SH-SY5Y建立神经细胞损伤模型。采用甲基噻唑基四唑染色法检测细胞活力,采用流式细胞仪检测细胞内活性氧(reactive oxygen species,ROS)水平,采用罗丹明123染色流式细胞仪分析SH-SY5Y细胞线粒体膜电位的改变,采用Western blot法分析线粒体细胞色素C的表达。结果 0.1～10μmol/L Pae可以剂量依赖性地抑制MPP+诱导的SH-SY5Y细胞的凋亡;10μmol/L Pae能够升高细胞线粒体膜电位,减少MPP+引起的细胞内ROS的产生,降低细胞色素C的表达。结论 Pae可防止MPP+损伤SH-SY5Y细胞,其保护作用可能与减少SH-SY5Y细胞内ROS生成,增强SH-SY5Y细胞清除自由基的能力以及提高SH-SY5Y细胞内线粒体膜电位水平有关。

Objective To investigate the mechanism by which paeonol （Pae） prevents 1-methyl-4 phenylpyridinium （MPP＋ ） induced damage in SH SY5Y cells. Methods Human neuroblastoma SH-Sx5Y cells were treated with MPP＋ in vitro to establish a nerve cell injury model. Cell viability was measured by MTT（methyl thiazolyl tetrazolium） assay, lntracellular reactive oxygen species （ROS） production was measured by flow cytometry （FCM）. The mitochondrial membrane potential of SH-SYSY cells was monitored by rhodamine 123 staining and FCM. The protein expression of cytoehrome C was determined by Western blot. Results Pae （0. 1-10 μmol/L） inhibited MPP＋-induced apoptosis of SH SY5Y cells in a dose-dependent manner. Pae （10μmol/L） increased mitochondrial membrane potential, reduced MPP＋-induced intracellular ROS production, and decreased the protein expression of cytochrome C in SH SY5Y cells. Conclusion Pae can protect SH-SY5Y cells from MPPI -induced damage, possibly by reducing in- tracellular ROS production, improving the capacity of scavenging free radicals, and increas;：g mitochondri- al membrane potential in SH-SY5Y cells.