熊胆粉对H_2O_2所致的PC12细胞凋亡的保护作用及机制  

Protective effects and mechanism of Bear Bile Powder on H_2O_2-induced apoptosis in PC12 cells

作　　者：富苏[1] 范吉平[2] 

机构地区：[1]中国中医科学院眼科医院,北京100040 [2]中国中医科学院,北京100700

出　　处：《中华中医药杂志》2014年第1期55-59,共5页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:探讨熊胆粉对H2O2所致的PC12细胞凋亡的保护作用及机制。方法:PC12细胞常规培养后,首先通过MTT检测细胞存活率的方法进行H2O2损伤模型的摸索和熊胆粉药物浓度的筛选;确定造PC12细胞损伤的H2O2浓度和熊胆粉的安全作用浓度后,将处于对数生长期的PC12细胞分为对照组、模型组和不同剂量熊胆粉组。用200μmol/L H2O2刺激PC12细胞使其发生凋亡,MTT法检测PC12细胞存活率、Hoechst33342荧光核染色观察细胞凋亡形态学改变、流式细胞仪检测PC12细胞凋亡率、Western blot方法检测与细胞凋亡线粒体途径密切相关的蛋白CytC和Caspase-3表达的变化。结果:200μmol/L H2O2诱导PC12细胞损伤明显,浓度小于等于100μmol/L的熊胆粉浓度对正常PC12细胞的存活率没有抑制作用。与模型组比较,熊胆粉组PC12细胞存活率显著提高(P<0.01),凋亡率显著下降(P<0.01),Cytc和Caspase-3的表达显著减少(P<0.01),其作用与剂量有关。结论:熊胆粉可抑制H2O2诱导的PC12细胞凋亡,其机制可能与抑制细胞凋亡线粒体途径中凋亡相关蛋白CytC和Caspase-3的表达有关。Objective： To investigate the protective effect and mechanism of Bear Bile Powder on H2O2-induced apoptosis in PC12 cells. Methods： PC12 cells were cultured, and the cell viability of PC12 cells injured by different H2O2 concentrations and proliferated by different Bear Bile Powders were observed by MTT assay to determine the concentration that will cause a H2O2-induced apoptosis model of PC12 cells and the security concentrations of Bear Bile Powder for PC12 cells. PC12 cells in the logarithmic growth phase were divided into control group, model group and low, medium and high dose Bear Bile Powder treatment groups. Apoptosis of PC12 cells were induced by H2O2 at the concentration of 200μmol/L, MTT assay was used to detect the cell viability of PC12 cells protected by Bear Bile Powder at different concentrations, Hoechst33342 fluorescent staining was used to observe morphological changes of cell apoptosis, the flow cytometry was used to detect the rate of apoptosis in PC12 cells, and the Western blot technology was used to detect the expressions of protein CytC and Caspase-3. Results： PC12 cells were injured by H2O2 at 200μmol/L concentration, while Bear Bile Powder at less than or equal to the concentration of100μmol/L did not inhibit the cell viability of PCI2 cells. Compared with the model group, Bear Bile Powder groups improved cell viability （P〈0.01）, decreased apoptotic cells and apoptosis rate （P〈0.01）, and decreased the expression of CytC and Caspase-3 （P〈0.01）, relating to doses of Bear Bile Powder. Conclusion： Bear Bile Powder can inhibit H2O2-induced apoptosis in PCI2 cells, while the mechanism may be related to the inhibition of the expression of apoptosis-related protein CytC and Caspase-3 in the mitochondrial pathway.

关 键 词：H2O2 PC12细胞 细胞凋亡 线粒体 熊胆粉 CYTC Caspase-3 

分 类 号：R285[医药卫生—中药学]