Wang Hao, Geng Zhao-Ming, Hu Zhi-Wei, Wang Shu-Yan, Zhao Bing
Department of Neurology, Tongde Hospital of Zhejiang Province, Hangzhou 310012, China.
Department of Clinical Medicine, Weifang Medical College, Weifang 261053, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2015 Jan;44(1):30-6. doi: 10.3785/j.issn.1008-9292.2015.01.005.
To investigate the effects of paeonol on neuron cell model of Parkinson disease (PD).
The cell model of Parkinson disease was induced by treatment of 1-Methyl-4-phenylpyridinium (MPP+) in PC12 cells, the PD model cells were treated with 1 μmol/L, 3 μmol/L or 9 μmol/L paeonol for 24h, respectively. Cell viability and LDH leakage were detected by MTT and lactate dehydrogenase (LDH) assay; the apoptosis of PC12 cells was assessed by Hoechst 33258 staining and flow cytometry; reactive oxygen species (ROS) production was detected by DCFH-DA method; and the ratio of Bax/Bcl-2 and activation of caspase-3 were determined by Western blotting.
MPP+ treatment significantly reduced cell viability, increased LDH leakage, enhanced the proportion of apoptotic cells and ROS production. In addition, MPP+ treatment dramatically increased the Bax/Bcl-2 ratio, and the activation of caspase-3. Compared to PD model group, paeonol treatment significantly enhanced cell viability, decreased LDH leakage, inhibited the proportion of apoptotic cells and ROS production, reduced the Bax/Bcl-2 ratio and the activated caspase-3 protein.
Paeonol can prevent PC12 cells from apoptosis induced by MPP+, and the mechanism may be associated with the down-regulation of ROS production, Bax/Bcl-2 ratio and Caspase-3 activation.
探讨丹皮酚对帕金森病(PD)神经元细胞模型的影响。
用1-甲基-4-苯基吡啶离子(MPP+)诱导PC12细胞建立帕金森病细胞模型,将PD模型细胞分别用1μmol/L、3μmol/L或9μmol/L丹皮酚处理24小时。采用MTT法和乳酸脱氢酶(LDH)检测法检测细胞活力和LDH泄漏;通过Hoechst 33258染色和流式细胞术评估PC12细胞凋亡情况;采用DCFH-DA法检测活性氧(ROS)生成;通过蛋白质免疫印迹法检测Bax/Bcl-2比值和半胱天冬酶-3的激活情况。
MPP+处理显著降低细胞活力,增加LDH泄漏,提高凋亡细胞比例和ROS生成。此外,MPP+处理显著增加Bax/Bcl-2比值和半胱天冬酶-3的激活。与PD模型组相比,丹皮酚处理显著提高细胞活力,降低LDH泄漏,抑制凋亡细胞比例和ROS生成,降低Bax/Bcl-2比值和激活的半胱天冬酶-3蛋白水平。
丹皮酚可防止MPP+诱导的PC12细胞凋亡,其机制可能与下调ROS生成、Bax/Bcl-2比值和半胱天冬酶-3激活有关。