人参皂苷Rg1通过抗氧化应激保护冈田酸诱导的PC12细胞损伤

摘要

目的体外细胞水平研究人参皂苷 Rg1在冈田酸(Okadaic Acid,OKA)诱导的阿尔茨海默病(Alzheimer’s dis-ease, AD)样神经毒性模型中的保护作用,并初步探索其作用机制。方法选用PC12细胞模拟神经元,采用OKA造模的同时给予Rg1(1、5、10μmol·L-1-),选用褪黑素(Melato-nin, Melat)10μmol·L-1-作为阳性对照。分别采用MTT、LDH法检测细胞存活率及死亡率,采用流式细胞术检测细胞凋亡情况,用 DCFH-DA 荧光探针法检测胞内 ROS 的堆积,并用试剂盒检测细胞内一系列抗氧化酶活性的变化。结果与对照组相比,OKA组细胞存活率明显降低、死亡率明显增加,细胞早期凋亡数量明显增多( P<0．01)；氧化应激相关指标检测发现, OKA 组胞内 ROS 堆积明显增多( P <0．01),总抗氧化能力(ABTS)明显降低(P<0．01),过氧化物酶(Catalase, CAT)(P<0．01),超氧化物歧化酶(Superox-ide Dismutase, SOD)、谷胱甘肽过氧化物酶( Glutathione per-oxidase,GSH-Px)活性都明显降低(P<0．05),GSSG/GSH比率明显升高(P<0．01)。与模型组相比,Rg1不同剂量组均能显著提高OKA模型中PC12细胞的存活率,降低细胞死亡率,并抑制胞内ROS水平、提高抗氧化酶活力增强抗氧化能力。结论人参皂苷Rg1在OKA诱导的AD样病理模型中可以通过抗氧化应激作用发挥抗细胞死亡、保护神经细胞的作用。%Aim To the investigate the protective effect of ginsenoside Rg1 in Alzheimer's disease ( AD)-like neurotoxicity model induced by okadaic acid ( OKA) in the cellular level , and explore the mecha-nism preliminarily. Methods The PC12 cells model, simulate neurons, induced by OKA was given Rg1 (1, 5,10 μmol·L-1), and melatonin (Melat) 10 μmol· L-1 was given as a positive control. MTT and LDH were carried out to assess the cell viability and mortality. To detect the accumulation of ROS, the DCFH-DA fluores-cent probe was conducted. And to assess the change of the activity of a variety of antioxidant enzymes, various kits were used, including ABTS、CAT、SOD、GSH-Px and GSSG/GSH. Results Compared with the control group, the survival rate of PC12 cell in OKA group re-duces significantly, the mortality rate was increased sig-nificantly , the number of early apoptotic cells was in-creased significantly (P<0. 01). Oxidative stress-relat-ed indicators show that ROS accumulation within the cells of OKA group increases significantly ( P<0. 01 ) , and the total antioxidant capacity ( ABTS ) decreases significantly ( P < 0. 01 ) , the activity of peroxidase (Catalase, CAT) (P <0. 01), glutathione peroxidase (glutathione peroxidase, GSH-Px) and superoxide dis-mutase ( superoxide dismutase, SOD) decreased signifi-cantly ( P <0. 05 ) , the rate of GSSG/GSH increased significantly ( P <0. 01 ) . Compared with the model group, the different doses of Rg1 could improve the sur-vival rate and decrease the mortality rate of PC12 cell significantly in the group of OKA, and could decrease the level of the accumulation of ROS, improve the activ-ity of antioxidant enzymes. Conclusion Ginsenoside Rg1 can decrease PC12 cell apoptosis by exerting an-tioxidant effects, and protect the nerve cells in AD-like pathology model induced by OKA.