槲皮苷对过氧化氢诱导PC12细胞损伤的保护作用

Protective effects of quercetin glycosides on PC12 cells injured by hydrogen peroxide

目的探讨槲皮苷对H2O2所致的PC12细胞凋亡的保护作用及机制。方法 PC12细胞培养后,MTT检测细胞存活率的方法进行H2O2损伤模型的摸索和槲皮苷药物浓度的筛选,将PC12细胞分为对照组、模型组和不同剂量槲皮苷组。用400μmol H2O2刺激PC12神经元细胞使其发生凋亡复制阿尔茨海默病(AD)模型,MTT法检测PC12细胞存活率、硫辛酰胺脱氢酶催化的INT显色反应检测乳酸脱氢酶(LDH)释放量和DAPI荧光核染色观察细胞凋亡形态学改变,Western blot方法检测Cytc和caspase-3表达的变化。结果 400μmol H2O2诱导PC12细胞损伤明显,与模型组比较,槲皮苷组PC12细胞存活率显著提高(P<0.01),凋亡率显著下降(P<0.01),LDH释放量和凋亡相关蛋白Cytc和caspase-3的表达显著减少(P<0.01)。结论槲皮苷可抑制H2O2诱导的PC12细胞凋亡,其机制可能与抑制细胞凋亡线粒体途径中凋亡相关蛋白Cytc和caspase-3的表达有关。

Objective To investigate the protective effects of quercitrin glycosides on PC12 cells injued by hydrogen peroxide. Methods PC12 cells were cultured,and the concentrations of H2O2 and quercitrin glycosides were explored by MTT,then the PC12 cells were divided into control group,model group and different doses of quercetin glycosides groups. In the study,the 400 μmol H2O2 was used to induce neuronal PC12 cells apoptosis to replicate the model of Alzheimer＇s disease（ AD）. Cell viability was detected by MTT assay,the reaction of diaphorase catalyzed INT chromogenic detected the lactate dehydrogenase（ LDH） content,and DAPI fluorescence staining was used to detect apoptotic nuclear,Western blot method to detect the expression of Cytc and caspase-3. Results 400 μmol H2O2 significantly induced the injury in PC12 cells,compared with the model group,PC12 cells survival rate significantly increased in quercitrin glycosides groups（ P〈0. 01）. The apoptosis rate was significantly decreased（ P〈0. 01）,the content of LDH was significantly lower（ P〈0. 01）and apoptosis-related proteins Cytc and caspase-3 were significantly reduced（ P〈0. 01） in quercitrin glycosides groups.Conclusion Quercitrin glycosides inhibit H2O2-induced PC12 cell apoptosis,the mechanism may be related to inhibit the expression of mitochondrial apoptosis pathway and apoptosis-related proteins Cytc and caspase-3.