目的 研究芍藥苷對(duì)過(guò)氧化氫（H₂O₂）誘導(dǎo)人神經(jīng)瘤母細(xì)胞（SH-SY5Y）凋亡的保護(hù)作用及其機(jī)制。方法 制備H₂O₂誘導(dǎo)的氧化應(yīng)激損傷模型。相差顯微鏡觀察SH-SY5Y細(xì)胞形態(tài)的變化；CCK-8法測(cè)定細(xì)胞存活率；Hoechst 33258染色法觀察細(xì)胞凋亡；碘化丙啶染色、流式細(xì)胞儀檢測(cè)細(xì)胞周期；2′, 7′-二氯熒光素二乙酸鹽（DCFH-DA）法檢測(cè)細(xì)胞內(nèi)活性氧（ROS）；INT顯色反應(yīng)法測(cè)定乳酸脫氫酶（LDH）的量；ELISA法檢測(cè)8-羥基脫氧鳥(niǎo)苷（8-OHdG）的量；caspase-3催化特異性底物反應(yīng)檢測(cè)caspase-3活性。結(jié)果 與對(duì)照組相比，200 μmol/L H₂O₂作用SH-SY5Y細(xì)胞24 h，使細(xì)胞存活力顯著下降（P＜0.01），細(xì)胞凋亡率上升（P＜0.01），增殖指數(shù)（PI）降低（P＜0.05），8-OHdG的量上升（P＜0.01），LDH釋放量和細(xì)胞內(nèi)ROS量增加（P＜0.01），caspase-3活性增強(qiáng)（P＜0.01）。與H₂O₂損傷組相比，芍藥苷終濃度為20～40 μmol/L，可濃度相關(guān)地減輕H₂O₂誘導(dǎo)的SH-SY5Y細(xì)胞上述指標(biāo)的變化（P＜0.05）；芍藥苷10 μmol/L組細(xì)胞PI、LDH和8-OHdG量未有明顯改觀，但能顯著減輕H₂O₂誘導(dǎo)的SH-SY5Y細(xì)胞毒性、ROS和caspase-3活性（P＜0.05）。結(jié)論 芍藥苷對(duì)H₂O₂誘導(dǎo)SH-SY5Y細(xì)胞損傷具有顯著保護(hù)作用，該作用可能與其清除ROS、減輕DNA氧化損傷、抑制細(xì)胞凋亡的caspase通路激活和調(diào)節(jié)細(xì)胞周期有關(guān)。

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.

國(guó)家自然科學(xué)基金資助項(xiàng)目（81274005）；河北省教育廳資助項(xiàng)目（2007302）