依达拉奉对淀粉样β蛋白25～35致PC12细胞氧化损伤的神经保护作用

Neuroprotective effects of edaravone on oxidative damage of PC12 cells induced by amyloid beta protein 25-35

目的:观察特异性清除羟自由基的神经保护剂依达拉奉对淀粉样β蛋白25～35诱导的PC12细胞氧化损伤的保护作用。方法:实验于2005-03/07在吉林大学中日联谊医院神经内科进行。将培养的PC12细胞分为3组:①正常对照组:加入含体积分数0.25的胎牛血清的DMEM维持液培养。②依达拉奉预处理组:加入依达拉奉20μmol/L预处理12h,然后加入淀粉样β蛋白25～3530μmol/L继续孵育24h。③淀粉样β蛋白25～35干预组:淀粉样β蛋白25～3530μmol/L孵育24h。采用MTT法测定细胞生存率;采用ELISA法测定羰基蛋白和晚期糖基化终产物的含量;硫代巴比妥酸法测定丙二醛的浓度。结果:①3组细胞生存率比较:正常对照组为(100.0±5.7)%,依达拉奉预处理组显著高于淀粉样β蛋白25～35干预组[(86.4±17.7)%,(42.5±9.4)%,P<0.001]。②3组细胞内羰基蛋白含量比较:正常对照组为(0.35±0.09)μmol/g,依达拉奉预处理组显著低于淀粉样β蛋白25～35干预组[(0.41±0.12),(0.81±0.17)μmol/g,P<0.01]。③3组晚期糖基化终产物水平比较:正常对照组为(12.21±3.26)mg/L,依达拉奉预处理组显著低于淀粉样β蛋白25～35干预组[(14.65±4.25),(26.57±5.18)mg/L,P<0.01]。④丙二醛浓度:正常对照组为(4.11±0.98)μmol/L,依达拉奉预处理组显著低于淀粉样β蛋白25～35干预组[(4.92±1.30),(7.58±1.01)μmol/L,P<0.01]。结论:依达拉奉能够减少淀粉样β蛋白25～35对PC12细胞内蛋白质氧化产物、晚期糖基化终产物及脂氧化终末产物的生成,具有神经保护作用。

AIM： To study the protective effect of edaravone, a kind of neuroprotective drug which specially scavenges hydroxy radical, on oxidative damage of PC12 cells induced by amyloid beta protein 25-35（Aβ25-35）. METHODS： The experiment was conducted at the Department of Neurology of China-Japan Hospital of Jilin University from March to July in 2005. Cultured PC12 cells were divided into three groups： ①Normal control group： PC12 cells were incubated in DMEM containing the fetal bovine serum with 0.25 volume fraction; ② Edaravone pre-treatment group： Edaravone at 20 μmol/L concentration was pre-processed for 12 hours, then 30 μmol/L Aβ25-35 was added in the culture solution for 24 hours. ③ Aβ25-35 intervention group： The Aβ25-35 at 30μmol/L concentration was incubated for 24 hours. Survival rate of PC12 cells was detected by MTY assays; The contents of protein carbonyl and advanced glycation end products （AGEs） were detected by ELISA, and the content of malondialdehyde （MDA） was detected by thiobarbituric acid method. RESULTS： ① Survival rate of PC12 cells in the three groups： That of the normal control group was （100.0±5.7）%, and that of the edaravone pretreatment group was significantly higher than that of Aβ25-35 intervention group [（86.4±17.7）%, （42.5±9.4）%, P〈0.001]. ② Contents of intracellular protein carbonyl： That of the normal control group was （0.35±0.09）μmol/g. It was lower in the edaravone pre-treatment group than that in the Aβ25-35 intervention group [（0.41＋0.12）,（0.81＋0.17） Ixmol/g,P 〈 0.01]. （~） Comparison of levels of AGEs and MDA： It was （12.21＋3.26） mg/L in the normal control group. It was lower obviously in the edaravone pre-treatmcnt group than that in the Aβ25-35 intervention group [（14.65±4.25）,（26.57±5.18）mg/L,P 〈 0.01]. ④ Concentration of MDA： It was （4.11±0.98）μmol/L in the normal control group. It was lower obviously in the edaravone pre-treatment group than that in the Aβ25-35 intervention group [（4.92±1.30）, （7.58±1.01） μmol/L,P〈0.01]. CONCLUSION： Edaravone is characteristic of neuroprotective effect, which can reduce the content of protein oxidative product, AGEs and lipid oxidative end product in the PC12 cells induced by Aβ25-35.