白藜芦醇对星形胶质细胞氧化损伤的保护作用  

Protective Effect of Resveratrol on Oxidative Injury in Astrocytes

作　　者：王景[1] 何超明[1] 

机构地区：[1]海南省农垦三亚医院神经内科,三亚572000

出　　处：《医药导报》2015年第8期1002-1006,共5页Herald of Medicine

基　　金：三亚市医疗卫生科技创新项目(YW1216)

摘　　要：目的探讨白藜芦醇(RES)对过氧化氢诱导星形胶质细胞氧化损伤的保护作用及其机制。方法星形胶质细胞传代培养,随机分为阴性对照组(以正常培养液培养),模型对照组(100μmol·L-1的过氧化氢作用12 h),RES小剂量组(20μmol·L-1RES孵育24 h后,加入过氧化氢作用12 h)和RES大剂量组(40μmol·L-1RES孵育24 h后,加入过氧化氢作用12 h)。采用MTT比色法测定细胞活力,流式细胞仪检测细胞凋亡率,Hochest33258染色观察凋亡细胞形态,比色法检测凋亡相关因子caspase-3及caspase-9的表达。结果 MTT结果显示5,10,20,40μmol·L-1RES孵育24 h后细胞活性分别为(100.46±3.17)%,(101.33±3.14)%,(101.33±1.30)%,(99.67±2.62)%,与阴性对照组(98.33±2.13)%比较,差异无统计学意义(P>0.05)。表明RES对星形胶质细胞活性无影响。用20及40μmol·L-1RES处理后,星形胶质细胞在过氧化氢作用12 h后引起损伤,其活性分别提高到(54.67±4.11)%和(70.33±2.61)%,与模型对照组比较,差异有统计学意义(t=3.59,7.13,P<0.01)。RES可以抑制过氧化氢诱导的细胞活性的下降。流式细胞计数结果显示用20,40μmol·L-1RES处理后,星形胶质细胞凋亡率分别下降到(35.51±3.56)%和(14.12±3.19)%,与模型对照组(46.31±4.16)%比较,差异有统计学意义(t=4.26,6.33 P<0.01)。Hochest33258染色结果显示RES可以抑制过氧化氢诱导的细胞凋亡。此外,RES还可以减少过氧化氢所致星形胶质细胞内caspase-3及caspase-9的表达,并且伴随RES作用时间的延长其表达量呈下降趋势。结论 RES通过抑制caspase-3及caspase-9的表达有效抑制了过氧化氢对星形胶质细胞的损伤,从而为其用于治疗中枢神经疾病提供实验依据。Objective To investigate the protective effect of resveratrol （RES） against hydrogen peroxide （ H2 02 ） - induced oxidative injury to astrocytes and the related mechanism. Methods Subcultured astrocytes were randomly divided into four groups： negative control group （treated with normal culture medium）, model control group （treated with 100 μmol·L^-1 H2O2 for 12 h） , resveratrol low dose group （treated with 20 μmol·L^-1 RES for 24 h H2O2 for 12 h） and rcsveratrol high dose group （ treated with 40 μmol·L^-1 RES for 24 h before H202 for 12 h）.Cell viability was detected by MTT assay, apoptosis rate was detected by flow cytometry, apoptotic cell morphology was detected by hochest33258 staining, and the expression of apoptosis-related factors such as caspase-3 and caspase-9 were measured by colorimetric detection. Results MTT assay showed that after treatment with 5, 10, 20, and 40 μmol·L^-1 RES for 24 h, cell viability was （100.46±3.17）%, （101.33±3.14） %, （101.33± 1.30）%, and （99.67±2.62）%, respectively, and the difference was not statistically significant compared with the negative control group [ （98.33±2.13）%, P〉0.05 ] .RES showed no effect on astrocyte activity. After treatment with 20 and 40 μmol·L^-1 RES, astrocyte activity was significantly elevated to （54.67±4.11）% and （70.33±2.61）% compared with model control group （t = 3.59, 7.13, P〈0.05 ）, RES inhibited hydrogen peroxide-induced decrease in cell viability. Flow cytometry results showed that after treatment with 20, 40 μmol·L^-1 RES, the apoptosis rate of astrocytes significantly decreased to （35.51±3.56）% and （14.12%±3.19）% （t=4.26, 6.33, P〈0.01） compared with model control group （46.31±4.16）%. Hochest 33258 staining showed that RES inhibited hydrogen peroxide-induced cell apoptosis, besides, the RES treatment also could reduce H2O2-induced expression of caspase-3 and caspase-9 in astrocytesin a time-dependent manner. Conclusion RES can inhibit hydrogen pe

关 键 词：白藜芦醇 星形胶质细胞 氧化损伤 

分 类 号：R286[医药卫生—中药学] R285.5[医药卫生—中医学]