李双月, 戚媛, 王哲敏, 陈若霖, 朴丰源. 2,5-己二酮对PC12细胞凋亡和线粒体膜电位的影响[J]. 环境与职业医学, 2015, 32(10): 966-969. DOI: 10.13213/j.cnki.jeom.2015.15199
引用本文: 李双月, 戚媛, 王哲敏, 陈若霖, 朴丰源. 2,5-己二酮对PC12细胞凋亡和线粒体膜电位的影响[J]. 环境与职业医学, 2015, 32(10): 966-969. DOI: 10.13213/j.cnki.jeom.2015.15199
LI Shuang-yue , QI Yuan , WANG Zhe-min , CHEN Ruo-lin , PIAO Feng-yuan . Effects of 2, 5-Hexanedione on Apoptosis and Mitochondrial Transmembrane Potential in PC12 Cells[J]. Journal of Environmental and Occupational Medicine, 2015, 32(10): 966-969. DOI: 10.13213/j.cnki.jeom.2015.15199
Citation: LI Shuang-yue , QI Yuan , WANG Zhe-min , CHEN Ruo-lin , PIAO Feng-yuan . Effects of 2, 5-Hexanedione on Apoptosis and Mitochondrial Transmembrane Potential in PC12 Cells[J]. Journal of Environmental and Occupational Medicine, 2015, 32(10): 966-969. DOI: 10.13213/j.cnki.jeom.2015.15199

2,5-己二酮对PC12细胞凋亡和线粒体膜电位的影响

Effects of 2, 5-Hexanedione on Apoptosis and Mitochondrial Transmembrane Potential in PC12 Cells

  • 摘要: 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& #177;2.41)%、(24.20& #177;4.07)%、(78.75& #177;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& #177;16.85)%、(51.44& #177;10.03)%、(22.81& #177;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。

     

    Abstract: Objective To explore the effects of 2, 5-hexanedione (HD) on apoptosis and mitochondrial transmembrane potential in PC12 cells and related potential mechanism.

    Methods HD (5, 10, and 20 mmol/L) were added into the culture medium of PC12 cells, and the non-exposed cells were set as control group. After 24 h, cell viability was tested by lactic acid dehydrogenase (LDH), cell morphological changes were evaluated after hematoxylin and eosin (HE) staining, apoptosis was determined by Hochest33258 staining and flow cytometric analysis with combination of Annexin V-FITC/PI staining, and mitochondrial membrane potential was tested by JC-1 dyeing.

    Results The cell viabilities were decreased to 92.23%, 86.40%, and 63.72% (all P < 0.05) of the control group after 5, 10, and 20 mmol/L HD administration, respectively, with blurred cell membrane and decreased cell count. Hochest33258 staining showed pyknotic and dark nucleus and strong blue fluorescence after HD exposure, displaying characteristic morphological changes of apoptosis. The results of flow cytometry showed that the apoptosis rates of the 5, 10, and 20 mmol/L HD groups were (10.62& #177;2.41)%, (24.20& #177;4.07)%, and (78.75& #177;8.39)% respectively, and all were significantly higher than that of the control group (0.3%) (P < 0.01). JC-1 dyeing showed that the mitochondrial transmembrane potentials of the 5, 10, and 20 mmol/L HD groups were (78.23& #177;16.85)%, (51.44& #177;10.03)%, and (22.81& #177;4.27)% of the control group, respectively.

    Conclusion HD could post toxic effects and induce remarkable apoptosis in PC12 cells and its mechanism may be associated with mitochondrial transmembrane potential decline.

     

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