金丝桃苷对甲基苯丙胺诱导的PC12细胞损伤的保护作用机制研究

Study on the protective mechanism of hyperoside on methamphetamine-induced PC12 cell injury

目的探讨金丝桃苷对甲基苯丙胺诱导的神经细胞损伤的影响及可能机制。方法建立甲基苯丙胺诱导的神经细胞损伤模型,使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法检测细胞存活率,彗星电泳法检测脱氧核糖核酸(DNA)损伤,通过检测神经细胞氧化应激相关指标及KELCH样ECH关联蛋白1(Keap1)/核因子E2相关因子2(Nrf2)通路蛋白表达变化,进一步探讨金丝桃苷产生保护作用的可能机制。结果与对照组比较,甲基苯丙胺组细胞存活率和抗氧化能力显著降低,DNA损伤程度和细胞凋亡增加,差异有统计学意义(P<0.05)。与甲基苯丙胺组比较,金丝桃苷干预组细胞抗氧化能力、胞核内Nrf2含量及其下游蛋白血红素加氧酶1和γ谷氨酰半胱氨酸合成酶的表达水平显著增高,细胞凋亡和DNA损伤程度显著降低,差异有统计学意义(P<0.05)。结论金丝桃苷能降低甲基苯丙胺诱导的神经细胞损伤,其保护作用机制可能与激活Keap1/Nrf2通路,促进Nrf2入核及其下游蛋白表达有关。

Objective To explore the effect of hyperoside on methamphetamine-induced nerve cell injury and its possible mechanism.Methods A methamphetamine-induced nerve cell injury model was established,and cell survival rate was detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide method,deoxyribonucleic acid(DNA)damage was detected using comet electrophoresis.Oxidative stress-related indicators in nerve cells and KELCH-like ECH associated protein 1(Keap1)/nuclear factor E2 associated factor 2(Nrf2)pathway protein expression changes were detected,and the possible mechanism of the protective effect of hyperoside was further explored.Results Compared with the control group,the survival rate and antioxidant capacity of cells in methamphetamine group decreased significantly,while the degree of DNA damage and apoptosis increased,with statistically significant difference(P<0.05).Compared with the methamphetamine group,the antioxidant capacity,Nrf2 levels in nucleus and the expressions of its downstream proteins heme oxygenase-1 andγ-glutamylcysteine synthetase were significantly higher,and the degree of apoptosis and DNA damage was significantly decreased in the hyperoside intervention group(P<0.05).Conclusion Hyperoside can reduce methamphetamine-induced nerve cell injury,and its protective mechanism may be related to activating Keap1/Nrf2 pathway and promoting nuclear translocation of Nrf2 and its downstream protein expression.