西红花酸对H_2O_2诱导PC12细胞损伤的保护作用

目的通过H2O2诱导PC12细胞损伤,建立氧化应激损伤的体外模型,以探讨西红花酸对细胞损伤的保护作用及其相关机制。方法观察不同浓度(0、50、100、200、300、400μmol/L)H2O2损伤PC12细胞12 h,用CCK-8检测细胞活力;不同浓度(0.1、1、5、10μmol/L)西红花酸预处理PC12细胞24 h,观察西红花酸对H2O2(200μmol/L)损伤细胞后的恢复作用,CCK-8检测细胞活力;罗丹明Rh123染色,流式检测线粒体膜电位(MMP);DCF-DA染色荧光照相术检测活性氧(ROS)水平;Western blot检测磷酸化ERK1/2的表达情况。结果 H2O2(0～400μmol/L)作用PC12细胞12 h后,细胞活力分别为(100±4.1)%、(102±1.9)%、(89±11.2)%、(52±2.6)%、(42±1.6)%、(8±0.4)%,细胞损伤呈明显的浓度依耐性;西红花酸预处理后,细胞活力由(45.12±3.15)%,分别上升为(51.88±4.24)%、(65.14±8.19)%、(57.66±5.58)%、(53.61±4.57)%;西红花酸(1μmol/L、5μmol/L)预处理组减少了线粒体膜电位(MMP)的下降,有效清除活性氧(ROS),激活磷酸化ERK1/2。结论上述实验表明西红花酸能对抗H2O2诱导的氧化应激损伤,表明西红花酸有抗氧化作用。

Protective effect of crocetin on hydrogen peroxide-induced cell injury

Objective To investigate the protective effect of crocetin in H2O2 induced rat pheochromocytoma line PC12,and explore the mechanisms on the injury.Methods The cell viability was detected by Cell Counting Kit-8(CCK-8) assay.Mitochondrial membrane potential(MMP) was stained by the Rhodamine123 and determined by flow cytometry.Reactive oxygen species(ROS) was observed by using DCF-DA staining and photofluorography.Phosphorylation of ERK1/2 was measured by Western blot.Results At the concentrations from 0 to 400 μmol/L for 12 h,the cell viability are(100±4.1)% 、(102±1.9)% 、(89±11.2)% 、(52±2.6)%、(42±1.6)%、(8±0.4)%,respectively.H2O2 dose-dependently inhibited cell viability.Exposure of the cells to different doses of crocetin(0～10 μmol/L) for 24 h followed by subsequent exposure to a single dose of H2O2(200 μmol/L),the cell viability increased from(45.12±3.15)% to(51.88±4.24)%、(65.14±8.19)%、(57.66±5.58)%、(53.61±4.57)%.Preincubation of cells with crocetin(1 μmol/L,5 μmol/L) 24 h prior to H2O2 exposure attenuated the decrease of MMP,scavenged ROS formation and activated ERK1/2 phosphorylation.Conclusion The crocetin holds potential for protective effects against H2O2-induced injury.