摘要
目的探讨活性氧(ROS)清除剂N-乙酰半胱氨酸(NAC)能否保护PC12细胞对抗化学性缺氧引起的损伤。方法应用化学性低氧模拟物氯化钴(CoCl2)处理PC12细胞,建立化学性缺氧损伤PC12细胞的实验模型。在CoCl2处理PC12细胞前60 min将NAC加入培养基中,作为预处理。应用CCK-8比色法检测细胞存活率;Hoechst33258核染色法观察细胞凋亡的形态学改变;罗丹明123(Rh123)染色荧光显微镜照像检测线粒体膜电位(MMP);Western blot法检测Caspase-3蛋白的表达水平。结果 600μmol/L CoCl2明显地损伤PC12细胞,表现为降低细胞存活率,增加凋亡细胞,激活Cas-pase-3及降低MMP。在CoCl2损伤PC12细胞前60 min,应用500μmol/L NAC作为预处理能明显地抑制CoCl2对PC12细胞的损伤作用,使细胞存活率显著升高,细胞凋亡率及Cleaved Caspase-3表达降低,并显著对抗CoCl2对MMP的抑制作用。结论NAC能明显地对抗化学性缺氧诱导的PC12细胞损伤,此保护作用与其改善MMP,抑制Caspase-3活化等机制有关。
Objective To investigate the protective effect of reactive oxygen species（ROS） scavenger,N-acetyl-L-cysteine（NAC）,against PC12 cells from injuries induced by chemical hypoxia.Methods PC12 cells were treated with cobalt chloride（CoCl2）,a chemical hypoxia-mimetic agent,to establish the chemical hypoxia-induced PC12 cells injury model.NAC was added into cell medium 60 min prior to CoCl2 exposure.The cell viability was evaluated using cell counter kit（CCK-8）.Morphological changes in apoptotic PC12 cells were detected by Hoechst 33258 staining and photofluorography.Mitochondrial membrane potential（MMP） was determined by Rhodamine123（Rh123） staining followed by photofluorography;the expression of cleaved caspase-3 was detected by western blot.Results Exposure of PC12 cells to 600 μmol/L CoCl2 for 24 h resulted in significantly cell injuries,evidenced by a decrease in cell viability,increases in apoptotic cells,and cleaved caspase-3 expression as well as loss of MMP.Pretreatment with NAC at the concentration of 500 μmol/L 60min before CoCl2 exposure inhibited CoCl2-induced PC12 cells injuries,leading to a significant increase in cell survival rate,decreases in apoptotic percent of PC12 cells and cleaved caspase-3 expression.Meanwhile,NAC attenuated the ameliorate MMP in PC12 cells.Conclusions NAC significantly protects PC12 cells against injury induced by chemical hypoxia,which is associated with decreasing caspase-3 activation and ameliorating MMP.
出处
《解剖学研究》
CAS
2011年第6期434-437,共4页
Anatomy Research
基金
广东省科技计划项目(2010B080701035)
