Aβ_(25-35)致PC12细胞核酸、蛋白质氧化损伤及依达拉奉的保护作用

Nucleic acid and protein oxidative damage of PC12 cells induced by Aβ_(25-35) and neuroprotective effect of Edaravone

目的研究依达拉奉对淀粉样β蛋白25-35(Aβ25-35)诱导的PC12细胞核酸、蛋白质氧化损伤的保护作用。方法实验对象分为三组:依达拉奉保护组(依达拉奉20μmol/L,Aβ25-3530μmol/L)、Aβ25-35干预组(Aβ25-3530μmol/L)和正常对照组。采用MTT法测定细胞生存率;慧星法测定DNA单链损伤;ELISA法测定羰基蛋白含量;比色法测定羟自由基(.OH)并计算.OH清除率。结果与正常对照组相比,Aβ25-35干预组细胞生存率降低,DNA单链损伤明显加重,细胞内羰基蛋白含量升高(P均<0.001)。依达拉奉保护组与Aβ25-35干预组相比,细胞生存率明显升高,DNA单链损伤明显减轻,细胞内羰基蛋白含量减低(P<0.001或P<0.01),但都未达到正常水平。依达拉奉对.OH的有效清除率可高达79.98%。结论依达拉奉能够通过清除.OH来减轻DNA损伤,并能减少细胞内蛋白质氧化产物的生成,具有神经保护作用。

Objective To study neuroprotective effect of Edaravone against nucleic acid and protein oxidative damage of PC12 cells induced by amyloid beta protein（Aβ）25-35. Methods Objectives were divided into three groups： the pre-treatment （Edaravone 20 μmol/L,Aβ25-35 30 μmol/L）, the intervention goup （Aβ25-35 30 μmol/L） and the normal group. Survival rate d PCI2 cells was detected by MTT assays, assessment of DNA damage by comet assay,and the content of protein carbonyl and hydroxy rascal（. OH） by chromatometry, and the clearance of. OHwas calculated. Results The content of intracellular protein carbonyl in the intervention group with Aβ25-35 were higher than those in the normal group, and DNA danage was significantly increased, so did survival rate of PC12 cell （ P 〈 0.001, all）. Compared with the intervention group with Aβ25-35, the content of intracellular protein carbonyl in the pre-treatment group with Edaravone were lowered and DNA damage was significantly lessened, but survival rate of PC12 cells was obviously incrcased （ P 〈 0.001, or P 〈 0.01）. The effective clearance rate of Edaravone to . OH reached up to 79. 98%. Conclusion Edaravone is characteristic of neuroprotective effect which it can reduce DNA injury and the content of protein oxidative product in the PC12 cells induced by Aβ25-35 through cleaning up hydroxy radical.