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[目的]探讨2,5-己二酮(HD)对PC12细胞的毒性作用及可能机制。[方法]分别用5、10、20 mmol/L HD处理PC12细胞24 h,用未暴露HD的细胞作为正常对照组。分别用乳酸脱氢酶法检测细胞存活率,苏木素-伊红染色检测细胞形态学变化,Hochest33258染色和Annexin V-FITC/PI双染流式检测细胞凋亡,JC-1染色检测线粒体膜电位。[结果]5、10、20 mmol/L HD暴露使PC12细胞的存活率下降,仅分别为对照组的92.23%、86.40%、63.72%(均P<0.05),细胞丧失正常形态,界限模糊,细胞数量明显减少。Hochest33258染色显示,HD暴露使细胞核固缩浓染,发出较强蓝色荧光,呈凋亡特征性形态。流式检测显示,5、10、20 mmol/L HD暴露组细胞的凋亡率分别为(10.62±2.41)%、(24.20±4.07)%、(78.75±8.39)%,与对照组(0.3%)相比,差异均有统计学意义(P<0.01)。JC-1染色显示,5、10、20 mmol/L HD组线粒体膜电位分别为对照组的(78.23±16.85)%、(51.44±10.03)%、(22.81±4.27)%(均P<0.01)。[结论]HD对PC12细胞具有明显的毒性作用,可诱导细胞凋亡,其机制可能与线粒体膜电位下降有关。
[Objective] To investigate the toxic effects of 2,5-hexanedione (HD) on PC12 cells and its possible mechanism. [Methods] The PC12 cells were treated with 5, 10, 20 mmol / L HD for 24 h, and the cells without HD exposure were used as the normal control group. Cell viability was detected by lactate dehydrogenase method. Morphological changes were detected by hematoxylin-eosin staining. Hochest33258 staining and Annexin V-FITC / PI double staining were used to detect cell apoptosis. Mitochondrial membrane potential was detected by JC-1 staining. [Results] The survival rate of PC12 cells was decreased by exposure to 5, 10 and 20 mmol / L of HD, which were only 92.23%, 86.40% and 63.72% in the control group (all P <0.05) The number of cells decreased significantly. Hochest33258 staining showed that HD exposure caused the nucleus to shrink and stain, giving rise to a stronger blue fluorescence with apoptotic features. The results of flow cytometry showed that the apoptotic rates of cells exposed to 5, 10 and 20 mmol / L of HD were (10.62 ± 2.41)%, (24.20 ± 4.07) and (78.75 ± 8.39)%, respectively, ), The differences were statistically significant (P <0.01). JC-1 staining showed that the mitochondrial membrane potentials in 5,10,20 mmol / L HD group were (78.23 ± 16.85)%, (51.44 ± 10.03)% and (22.81 ± 4.27)% in the control group (all P <0.01) . [Conclusion] HD has obvious toxic effects on PC12 cells and induces apoptosis, which may be related to the decrease of mitochondrial membrane potential.