依达拉奉在庆大霉素诱导MDCK细胞凋亡中的保护作用

Protection of edaravone on apoptosis induced by gentamicin in MDCK cells

为研究依达拉奉(edaravone,EDa)在庆大霉素(gentamicin,GM)诱导MDCK细胞凋亡中的保护作用,本研究用4 mmol/L GM和40?mol/L EDa单独或联合处理MDCK细胞24 h,Annexin V-FITC/PI双染检测细胞凋亡率;Hochest 33258染色观察细胞核形态变化;比色法检测总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-PX)、过氧化氢酶(CAT)活力和丙二醛(MDA)含量;荧光染色检测活性氧(ROS)含量;Western-blot检测Bax与Bcl-2表达量,caspase-9、caspase-3和PARP蛋白活化情况,细胞色素C(Cyt C)和凋亡诱导因子(AIF)释放情况。结果显示,与对照组相比,EDa组无显著性影响。与GM组相比,GM+EDa组细胞凋亡率显著或极显著性下降(P<0.05或P<0.01);T-SOD和GSH-PX活力显著性下降(P<0.05);CAT活力极显著性上升(P<0.01);MDA含量极显著性下降(P<0.01);ROS含量显著性下降(P<0.05);Bcl-2/Bax比值显著性升高(P<0.05);caspase-9、caspase-3和PARP蛋白的活化水平显著或极显著性降低(P<0.05或P<0.01);Cyt C与AIF含量显著性降低(P<0.05)。结果表明,依达拉奉在庆大霉素通过线粒体凋亡途径诱导MDCK细胞凋亡中起保护作用。

In order to investigate the protective machanism of edaravone(EDa)on apoptosis induced by gentamicin(GM)in MDCK cells,MDCK cells were treated with 4 mmol/L GM and 40?mol/L EDa for 24 h respectively or unitedly.The apoptotic rate was detected by Annexin V-FITC/PI double staining.The changes of nuclear morphology were observed by Hochest 33258 staining.T-SOD,GSH-PX,CAT activity and MDA content were detected by colorimetric assay.ROS content was detected by fluorescent staining.The expression of Bax and Bcl-2,the activation of caspase-9,caspase-3 and PARP protein,the release of Cytochrome(Cyt C)and apoptosis-induced factor(AIF)were detected by Western-blotting.In result,the Eda group was no significant effect compared with the control group.Compared with the GM group,the apoptotic rate of the GM+EDa group was significantly or extremely ignificantly decreased(P<0.05 or P<0.01).The T-SOD and GSH-PX activities were significantly decreased(P<0.05).CAT activity was extremely significantly increased(P<0.01).MDA content decreased extremely significantly(P<0.01).ROS content decreased significantly(P<0.05).Bcl-2/Bax ratio increased significantly(P<0.05).The activation level of caspase-9,caspase-3 and PARP protein was significantly or extremely significantly decreased(P<0.05 or P<0.01).The contents of Cyt C and AIF were significantly decreased(P<0.05).These findings suggested that EDa can relieve oxidative damage to supperess the gentamicin-induced apoptosis through mitochondrial apoptotic pathway.