氧化还原因子1对体外培养大鼠耳蜗螺旋神经节氧化损伤的保护作用

目的 观察表达无嘌呤无嘧啶核酸内切酶/氧化还原因子1(apurinic/apyrimidimicendonuclase/redox factor 1,APE/Ref-1)的重组腺病毒感染对H2O2所致体外培养大鼠耳蜗螺旋神经节细胞氧化损伤的保护作用.方法 体外培养大鼠耳蜗螺旋神经节细胞,APE/Ref-1腺病毒表达载体感染48 h后,加入不同浓度H2O2(0、10、25、50、100及300 μmol/L)干预1 h,更换正常培养液后继续培养24 h,通过蛋白免疫印迹分析、四甲基偶氮唑蓝(MTT)法、原位缺口末端标记法(TUNEL)分别检测感染后螺旋神经节细胞APE/Ref-1蛋白表达、细胞活力以及凋亡情况.结果 通过腺病毒感染实现了APE/Ref-1基因在体外培养耳蜗螺旋神经节细胞的过表达,H2O2浓度为50～300 μmol/L时,APE/Ref-1组同对照组比较,细胞活力提高、凋亡率降低(P值均<0.01).结论 腺病毒介导的APE/Ref-1过表达对H2O2所致螺旋神经节细胞氧化损伤具有保护作用.

Adenovirus-mediated APE/Ref-1 expression protects rat spiral ganglion cells from oxidative damage

Objective To address the question if apurinic/apyrimidimic endonuclase/redox factor 1 (APE/Ref-1) involved in preventing spiral ganglion cells oxidative damage after oxidative stress. Methods Primary cultured rat spiral ganglion cells were infected with the adenovirus containing APE/Ref-1 for 48 h, then treated with H2O2 (0, 10,25,50,100,300 μmol/L)for 1 h, and finally changed back into normal medium. Western blot were used to detect the level of APE/Ref-1 protein in the infected cells to ensure APE/Ref-1 over expression as a result of adenovirus infection. The cell viability was determined by MTF and the apoptosis of spiral ganglion cells was determined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). Results Western blot showed that infection of adenovirus resulted in APE/Ref-1 over expression in the spiral ganglion cells. Over expression of APE/Ref-1 significantly improved cell viability in cultures treated with different concentration H2O2 from 50 to 300 μmol/L However, the apoptosis of cells was significantly inhibited. Conclusions Overe xpression of APE/Ref-1 could protect spiral ganglion cells from oxidative damage.