摘要
目的探讨线粒体ATP敏感钾通道(K^+-ATP)开放剂——二氮嗪对氧糖剥夺PC12细胞的保护作用及其机制。方法体外培养PC12细胞,分为正常对照组、氧糖剥夺组、二氮嗪预处理组(氧糖剥夺前20 min给予二氮嗪200μmol/L)。以改良噻唑蓝法测定细胞活性,采用Heochst33342/PI双染法测定细胞凋亡率,Fluo-3/AM(钙离子荧光探针)检测PC12细胞内钙离子的浓度。结果①与正常对照组比较,氧糖剥夺后细胞活力明显降低(P<0.01);在氧糖剥夺前给予200μmol/L的二氮嗪可使细胞活力明显升高(P<0.01)。②正常对照组细胞凋亡率为(7.5±2.1)%,氧糖剥夺组为(42.0±2.6)%,二氮嗪预处理组为(21.3±3.2)%。与正常对照组比较,氧糖剥夺组和二氮嗪预处理组差异均有统计学意义(P<0.01),二氮嗪预处理组与氧糖剥夺组比较,差异亦有统计学意义(P<0.01)。③正常对照组、氧糖剥夺组、二氮嗪预处理组细胞内游离钙离子平均荧光值分别为(120.6±1.6)%,(280.8±2.5)%,(167.0±3.9)%。与正常对照组比较,氧糖剥夺组和二氮嗪预处理组差异均有统计学意义(P<0.01),二氮嗪预处理组与氧糖剥夺组比较,差异亦有统计学意义(P<0.01)。结论线粒体K^+-ATP开放剂——二氮嗪可增强氧糖剥夺细胞的活力,减少细胞的凋亡率,其机制可能与降低细胞内钙超载有关。
Objective To investigate the protective effect of mitochondrial ATP-sensitive potassium channel （K＋-ATP） opener diazoxide on oxygen glucose deprivation-induced PC12 cells and its mecha- nisms. Methods Pheochromocytoma （PC12） cells cultured in vitro were divided into normal control group, oxygen-glucose deprivation group, and diazoxide pretreatment group （diazoxide 200μmol/L was given before oxygen-glucose deprivation）. The cell activity was measured by the modified MTF assay. Ho- echst 33342/PI double staining was used to detect apoptosis rate. Fluo-3/AM （calcium fluorescence probe） was used to detect the concentration of calcium in PC12 cells. Results （1）Compared to the normal control group, the cell viability decreased significantly after oxygen-glucose deprivation （ P 〈 0.01 ）. Giving diazoxide （200 μmol/L） before oxygen-glucose deprivation could significantly increase cell viability （P 〈0.01 ）. （2）The apnptosis rate in the normal control group was 7.5 ± 2. 1%, in the oxygen-glucose deprivation group was 42.0 ± 2. 6% , and in the diazoxide pretreatment group was 21.3 ± 3.2%. Compared to the normal control group, there was statistical difference between the oxygen-glucose deprivation group and the diazoxide pretreatment group （ P 〈 0.01 ）. There was also statistical difference between the diazoxide pretreatment group and the oxygen-glucose deprivation group. （3）The mean fluorescence values of intracellular free calcium ions in the normal control group, oxygen-glucose deprivation group, and diazoxide pretreatment group were 120. 6± 1.6% , 280. 8± 2.5% , and 167.0 ± 3.9% , respectively. Compared to the normal control group, there were statistical significance between the oxygen-glucose deprivation group and diazoxide pretreatment group （ P 〈 0.01 ）. There were also statistical significance between the diazoxidc pretrcatment group and the oxygen-glucose deprivation group （P 〈 0.01 ）. Conclusion K ＋ -ATP opener can enhance the viability of ox
出处
《中国脑血管病杂志》
CAS
2010年第11期592-595,共4页
Chinese Journal of Cerebrovascular Diseases
基金
辽宁省科学技术计划资助项目(No 2008225022)
