核桃多肽对过氧化氢及β淀粉样肽25-35诱导PC12细胞氧化损伤的影响

Effects of Walnut Polypeptides on Oxidative Damage of PC12 Cells Induced by H_2O_2 and Aβ25-35

目的:探讨核桃多肽对过氧化氢(H_2O_2)及β淀粉样肽25-35(Aβ25-35)导致的PC12细胞损伤的保护作用。方法:以H_2O_2及Aβ25-35损伤PC12细胞建立氧化应激损伤模型,以不同浓度核桃多肽(0.1,0.25,0.5,0.75,1,2 mg·L^(-1))预保护PC12细胞,采用MTT法检测细胞活力;采用乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GSH-Px)等试剂盒检测H_2O_2及Aβ25-35对PC12细胞的氧化损伤,并研究核桃多肽的抗氧化作用;采用DCFH-DA结合荧光分光光度计检测细胞内的活性氧(ROS)水平。结果:与模型组相比,核桃多肽呈剂量依赖性地提高H_2O_2及Aβ25-35诱导的PC12细胞存活率;核桃多肽组的抗氧化酶SOD、CAT和GSH-Px活性明显提高,MDA和细胞内的ROS水平明显降低(P<0.05或P<0.01)。结论:核桃多肽对H_2O_2及Aβ25-35导致的PC12细胞损伤具有良好的保护作用,其作用机制可能与提高PC12细胞抗氧化能力有关。

Objective: To investigate the protective effect of walnut peptide on the damage of PC12 cells induced by β-amyloid protein 25-35( Aβ25-35) and hydrogen peroxide( H2O2). Methods: PC12 cells were injured by H2O2 and Aβ25-35 to establish oxidative stress injury model and different concentrations of walnut polypeptide( 0. 1,0. 25,0. 5,0. 75,1 and 2 mg·L^-1) were administrated to provide pre-protection for PC12 cells. The cell viability was measured by MTT. Lactate dehydrogenase( LDH),superoxide dismutase( SOD),malondialdehyde( MDA),catalase( CAT) and glutathione peroxidase( GSH-Px) kits were used to detect the oxidative damage of H2O2 and Aβ25-35 on PC12 cells,and study the antioxidant effect of walnut polypeptide. The intracellular reactive oxygen species( ROS) levels were detected by DCFH-DA combined with a fluorescence spectrophotometer. Results: Compared with the model group,walnut polypeptide increased the survival rate of PC12 cells induced by H2O2 and Aβ25-35 in a dose-dependent manner,the activities of antioxidant enzymes SOD,CAT and GSH-Px in walnut polypeptide group were significantly increased,and the levels of MDA and ROS in cells were significantly increased( P < 0. 05 or P < 0. 01). Conclusion: Walnut polypeptide has a good protective effect on PC12 cells damaged by H2O2 and Aβ25-35,which may be related to its anti-oxidant action.