目的 研究芍药苷对过氧化氢（H₂O₂）诱导人神经瘤母细胞（SH-SY5Y）凋亡的保护作用及其机制。方法 制备H₂O₂诱导的氧化应激损伤模型。相差显微镜观察SH-SY5Y细胞形态的变化；CCK-8法测定细胞存活率；Hoechst 33258染色法观察细胞凋亡；碘化丙啶染色、流式细胞仪检测细胞周期；2′, 7′-二氯荧光素二乙酸盐（DCFH-DA）法检测细胞内活性氧（ROS）；INT显色反应法测定乳酸脱氢酶（LDH）的量；ELISA法检测8-羟基脱氧鸟苷（8-OHdG）的量；caspase-3催化特异性底物反应检测caspase-3活性。结果 与对照组相比，200 μmol/L H₂O₂作用SH-SY5Y细胞24 h，使细胞存活力显著下降（P＜0.01），细胞凋亡率上升（P＜0.01），增殖指数（PI）降低（P＜0.05），8-OHdG的量上升（P＜0.01），LDH释放量和细胞内ROS量增加（P＜0.01），caspase-3活性增强（P＜0.01）。与H₂O₂损伤组相比，芍药苷终浓度为20～40 μmol/L，可浓度相关地减轻H₂O₂诱导的SH-SY5Y细胞上述指标的变化（P＜0.05）；芍药苷10 μmol/L组细胞PI、LDH和8-OHdG量未有明显改观，但能显著减轻H₂O₂诱导的SH-SY5Y细胞毒性、ROS和caspase-3活性（P＜0.05）。结论 芍药苷对H₂O₂诱导SH-SY5Y细胞损伤具有显著保护作用，该作用可能与其清除ROS、减轻DNA氧化损伤、抑制细胞凋亡的caspase通路激活和调节细胞周期有关。

Objective To investigate the neuroprotective effects of paeoniflorin (PF) against cell death induced by hydrogen peroxide (H₂O₂) in human neuroblastoma cells and its mechanisms. Methods H₂O₂ was used to induce SH-SY5Y cell damages, and the cell survival rate was detected by CCK-8 assay; the cell morphologic changes were observed by inverted optical microscope; the apoptosis was tested using Hoechst 33258 staining; flow cytometer (FCM) and propidium iodide staining were used to analyze the apoptosis and cell cycle alteration; reactive oxygen species (ROS) production was determined by 2′, 7′-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescence; lactic dehydrogenase (LDH) release was detected by reaction of diaphorase and INT; 8-OHdG production was determined by enzyme-linked immunosorbent assay (ELISA); caspase-3 activity was determined by caspase-3 catalyzing the specific substrate. Results Compared with control group, after the treatment with H₂O₂ (200 μmol/L) for 24 h, the viability and proliferation index of CH-SY5Y cells were significantly decreased (P < 0.01, 0.05), the apoptosis rate and content of 8-OHdG were increased (P < 0.01), the LDH resease and ROS production were increased (P < 0.01); the activity of caspase 3 was increased (P <0.01). Compared with H₂O₂ injury group, PF (20—40 μmol/L) significantly ameliorated the changes in SH-SY5Y cells induced by H₂O₂ in concentration-related manner (P < 0.05). PF (10 μmol/L) did not significantly change the above indexes except the cell viability, ROS, and caspase-3 activity induced by H₂O₂ (P < 0.05). Conclusion PF has the significant protective effect against the H₂O₂-induced cell injury, which may be related to eliminatinging ROS, alleviating DNA oxidative damage, regulation of cell cycle, and inhibition of apoptosis of caspase pathway activation.

国家自然科学基金资助项目（81274005）；河北省教育厅资助项目（2007302）