目的以过氧化氢(H2O2)为工具药,建立阿尔茨海默病(AD)的氧化应激模型。观察雷公藤内酯醇(T10)的保护作用并初步探讨其作用机制。方法用不同浓度(25、50、100、200μmol/L)的H2O2处理PC12细胞24h,100μmol/LH2O2处理PC12细胞不同时间(6、12、24及48h),建立细胞损伤模型。用1、10、30nmol/LT10预孵育PC12细胞12h后加入100μmol/LH2O2共同作用24h以探讨T10的保护作用。MTT法检测细胞存活率,化学比色法测定乳酸脱氢酶(LDH)释放量,并采用荧光素酶报告基因方法检测转录因子NF-κB转录活性以探讨T10作用机制。结果随着H2O2浓度增加或作用时间延长,PC12细胞MTT值逐渐降低,而LDH释放量逐渐增加。100μmol/LH2O2处理细胞24h,MTT值明显下降,LDH释放量增加。而1、10、30nmol/L的T10预孵育12h可明显减轻PC12细胞的损伤,提高MTT值并降低LDH释放量。100μmol/LH2O2作用4h可明显增加PC12细胞NF-κB的转录活性,而1、10、30nmol/L的T10预处理12h可显著拮抗NF-κB转录活性的升高。结论T10可以有效的拮抗H2O2对PC12细胞的氧化损伤,作用机制可能与其降低转录因子NF-κB的转录活性有关。 Objective To establish an oxidative stress model of Alzheimer’s disease (AD) using hydrogen peroxide (H2O2) as a drug. The protective effect of triptolide (T10) was observed and its mechanism of action was initially explored. Methods PC12 cells were treated with different concentrations (25, 50, 100, 200 μmol/L) of H2O2 for 24 h, and PC12 cells were treated with 100 μmol/L H2O2 for different times (6, 12, 24, and 48 h) to establish a cell injury model. PC12 cells were preincubated with 1, 10, and 30 nmol/LT10 for 12 h and 100 μmol/L H2O2 was added for 24 h to investigate the protective effect of T10. The survival rate of cells was determined by MTT assay, and the release of lactate dehydrogenase (LDH) was determined by chemical colorimetry. The transcriptional activity of transcription factor NF-κB was detected by luciferase reporter gene method to explore the mechanism of T10 action. Results With the increase of H2O2 concentration or the prolonged action time, the MTT value of PC12 cells gradually decreased, while the release of LDH increased gradually. 100μmol/LH2O2 treatment of cells for 24h, MTT value decreased significantly, LDH release increased. Pre-incubation of 1, 10, and 30 nmol/L T10 for 12 h could significantly reduce the damage of PC12 cells, increase the MTT value and decrease the release of LDH. 100μmol/LH2O2 treatment for 4h significantly increased the transcription activity of NF-κB in PC12 cells, while 1,10, 30nmol/L T10 pretreatment for 12h significantly antagonized the increase of NF-κB transcription activity. Conclusion T10 can effectively antagonize the oxidative damage of PC12 cells induced by H2O2. Its mechanism may be related to the decrease of transcriptional activity of transcription factor NF-κB.