5-取代-1-氮杂蒽醌类衍生物A7对过氧化氢诱导转染人APP695基因的SH-SY5Y细胞应激损伤的保护作用

Protective effects of 5-substituted-1-aza-anthraquinone derivative A7 on stress injury induced by hydrogen peroxide in SH-SY5Y cells transfected with human APP695 gene

目的探究5-取代-1-氮杂蒽醌类衍生物A7对过氧化氢诱导转染人APP695基因的SH-SY5Y细胞(APPsw细胞)氧化应激损伤的保护作用。方法将APPsw细胞传代培养24 h后,加入不同浓度(0.01,0.1,1.0和10μmol·L^-1)的A7预处理24 h,再加入200μmol·L^-1过氧化氢(H 2 O 2)继续培养24 h,用MTT法检测细胞活性,用Hoechst染色法检测细胞凋亡,用Annexin V/PI双染流式细胞术检测细胞凋亡率。结果5-取代-1-氮杂蒽醌类衍生物A7可剂量依赖性地增加过氧化氢处理后的细胞活力,经不同浓度的A7预处理后,加过氧化氢处理组的细胞活性明显低于对应单独A7处理组的细胞活性,但均高于单独过氧化氢处理组的细胞活性,差异具有统计学意义(P<0.05)。经不同浓度A7预处理后,加过氧化氢处理组与过氧化氢组比较细胞核中荧光显著变暗。过氧化氢组细胞早凋率为42.4%,A7(0.01,0.1,1.0和10μmol·L^-1)预处理24 h后加过氧化氢处理组细胞早凋率分别为22.7%,19.9%,16.4%和15.9%,差异具有统计学意义(P<0.05)。结论5-取代-1-氮杂蒽醌类衍生物A7对过氧化氢诱导APPsw细胞的氧化应激损伤具有保护作用,可抑制细胞凋亡。

Objective To investigate the protective effect of 5-substituted-1-aza-anthraquinone derivative A7 on oxidative stress injury induced by H2O2 in SH-SY5Y cells transfected with human APP695 gene(APPsw cells).Methods After 24 h subculturing,APPsw cells were preprocessed respectively with different concentrations(0.01,0.1,1.0和10μmol·L^-1)of A7 for 24 h,and then hydrogen peroxide(H2O2 200μmol·L^-1)was added to the culture medium.After 24 h incubation,the cell viability was detected by MTT,and the apoptosis rate was analyzed by Hoechst staining and Annexin V/PI double-staining flow cytometry.Results The cell viability after being processed with different concentrations of A7 derivatives and H2O2 was significantly lower than that after being processed with corresponding single A7 derivative,both of which were higher than that being processed with H2O2 alone(P<0.05).The fluorescence in cell nucleus processed by different concentration of A7 and H2O2 was significantly darker than that processed by H2O2 alone.The apoptosis rate in H2O2 group was 42.4%,while the apoptosis rates in preprocess with A7 derivatives(0.01,0.1,1.0 and 10μmol·L^-1)for 24 h and H2O2 were 22.7%,19.9%,16.4%and 15.9%,respectively(P<0.05).Conclusion The pre-administration of 5-substituted-1-aza-anthraquinone derivative A7 can effectively protect APPsw cells against H2O2-induced oxidative stress and inhibit cell apoptosis.