二甲基甲酰胺致人肝细胞凋亡及其对Bcl-2及Bax和Caspase-3表达的影响  

DMF-induced adult hepatocyte apoptosis and its effects on expression of Bcl-2, Bax and Caspase-3

作　　者：路艳艳[1,2] 宣志强[3] 李雪芝[1] 杨锦蓉[3] 王菁[1] 

机构地区：[1]浙江大学医学院环境医学研究所,杭州310058 [2]浙江省医学科学研究院 [3]浙江省疾病控制中心

出　　处：《中华劳动卫生职业病杂志》2008年第5期276-279,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：浙江省医药卫生科学研究基金（2005B019）

摘　　要：目的初步探讨二甲基甲酰胺（DMF）对正常成人肝细胞的致凋亡作用和对Bcl-2、Bax、Caspase-3表达的影响。方法以终浓度为0、50、100、150、200mmol／L的DMF作用于正常成人肝细胞12h后，应用流式细胞术和western blotting法检测细胞凋亡率及Bcl-2、Bax和Caspase-3表达的变化。结果与阴性对照组比较，各DMF剂量组凋亡率明显上升，差异有统计学意义（P〈0．01）。随着染毒剂量的增加，肝细胞凋亡率上升，呈现剂量．反应关系。且染毒后随着DMF剂量的增加，Bcl-2表达水平下降，差异有统计学意义（P〈0．01）。与对照组比较，Bax表达水平无明显变化，Bcl-2／Bax比值随着DMF剂量增加而下降，200mmol／L剂量组与阴性对照组比较，差异有统计学意义（P〈0．01）。在DMF150、200mmol／L剂量组可见到Caspase-3切割活化。结论DMF可诱导肝细胞发生凋亡，其机制可能与Bcl-2／Bax比值表达下调、Caspase-3发生切割活化有关。Objective To investigate the apoptosis-inducing effect of DMF on the human liver cells （HL-7702） in vitro. Methods Liver cells were exposed to different concentrations of DMF （0,50,100,150,200 mmol/L） for 12 hours. Apoptotic rate,the expression of Bax,Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively. Results The increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group （P〈0.01） ,the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance （P〉0.05）, as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control（P〈0.01 ）. The new lands of procaspase-3 in 150,200 mmol/L were observed,which demonstrated that there was active caspase-3. Conclusion DMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.

关 键 词：二甲基甲酰胺 肝细胞凋亡 BCL-2 BAX Caspase-3 

分 类 号：R285.5[医药卫生—中药学]