雷公藤单体T10对过氧化氢所致PC12细胞损伤的保护作用及其机制的研究

Neuroprotective effect of Tripterygium Wilfordii Hook F monomer T10 agaist hydrogen peroxide-induced PC12 cell damage and its mechanism

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 study the neuroprotective effect and its mechanism of Tripterygium Wilfordii Hook F monomer T10 against hydrogen peroxide （H2O2） -induced PC 12 cell damage. Methods The PC12 cell damage model was induced by treating PC12 cells with different concentrations （25, 50, 100, 200 μmol/L） of H2O2 for 24 h and with 100 μmol/L H2O2 for different time course（6, 12, 24, 48 h）. To explore the neuroprotective effects of T10, the cells were pretreated with 1, 10, 30 nmol/L T10 for 12 h, followed by co-treatment with 100 μmol/L H2O2 for 24 h. The viability of PC12 cells was detected by MTT conversion and LDH leakage. Furthermore, we measured the transcription activity of NF-κB by luciferase reporter gene assays to understand the underlying mechanism of the neuroprotective effects of T10. Results After being treated with different concentrations of H2O2 for 24 h or with 100 μmol/L H2O2 for different time course, the cell viability of PC12 cells decreased significantly. 100 μmol/L H2O2 treatment for 24 h induced a significant decrease in MTT metabolic rate and an increase in LDH leakage rate in the PC12 cells. 1, 10, 30 nmol/L T10 pretreatment inhibited H2O2-induced cell death and significantly inhibited the transcription activity of NF-κB induced by H2O2. Conclusion T10 can protect PC 12 cell fi＇om the death induced by H2O2, and the underlying mechanism may be involved in the capacity of T10 to inhibit the transcription activity of NF-κB.