二甲基甲酰胺对人肝细胞凋亡相关蛋白影响  

Effects of N,N-dimethylformamide on Caspase-3's activation in human hepatocytes in vitro

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

机构地区：[1]浙江省疾病预防控制中心辐射与核安全防护所,杭州310051 [2]浙江省医学科学院卫生学研究所 [3]浙江大学医学院环境医学研究所

出　　处：《中国公共卫生》2010年第11期1395-1397,共3页Chinese Journal of Public Health

基　　金：浙江省医药卫生科学研究基金项目(2005B019)

摘　　要：目的探讨不同剂量二甲基甲酰胺(DMF)在不同染毒时间下对人正常离体肝细胞半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)激活的影响。方法以0,6.25,25,100 mmol/L的DMF染毒正常成人肝细胞12 h,100 mmol/L的DMF染毒肝细胞0,3,6,12 h,应用蛋白印记法(Western法)观察Caspase-3激活的变化。结果随着染毒剂量的增加,Caspase-3原酶形式(Procapase-3)表达水平呈下降趋势,但各染毒组间比较,差异无统计学意义(P>0.05);100 mmol/L染毒组Caspase-3激活水平最高,灰度比值为0.55,各染毒组间比较差异有统计学意义(F=7.684,P<0.01);随着染毒时间增加,Caspase-3激活水平逐渐增加,12 h染毒组灰度比值最高,达0.48,各组间比较,差异有统计学意义(F=9.351,P<0.01)。结论 Caspase-3的激活参与了DMF致人肝细胞的凋亡过程,并呈现良好的时间-剂量效应关系。Objective To discuss the effect of N,N-dimethylformamide（DMF） at different dose and exposure time on Caspase-3＇s activation in normal human liver cells in vitro.Methods The human hepatocytes were exposed to DMF at the concentrations of 6.25,25,100 mmol/L for 12 hours.Then the hepatocytes were exposed to DMF at the concentration of 100 mmol/L for 0,3,6 and 12 hours.The control group was treated with phosphate buffered saline.Western blotting was performed to observe the activation of Caspase-3 in human hepatocytes.Results With the increasing of DMF dose,the ratio of pro-Caspase-3＇s grey value showed a declined trendency on the whole but without statistical difference between different groups（F = 1.363,P 0.05）.Different dose of DMF could increase the activation of Caspase-3（F = 7.684,P 0.01） with a tendency of increment with the increasing of dose.The average ratio of Caspase-3 subunits＇ grey value in 100 mmol/L group was the highest（0.55）.With the increasing of exposure time,there was no statistical difference in pro-Caspase-3＇s grey value between different groups（F = 0.490,P 0.05） with a declined trendency as the increasing of exposure.The ratio of Caspase-3 subunits＇ grey value significantly increased with the time of exposure（F = 9.351,P 0.01） and the highest was 0.48 in the group of 12 hours＇ exposure.Conclusion Activation of Caspase-3 involves in the process of apoptosis induced by DMF in human hepatocytes and the activation level of Caspase-3 activation shows a good time-and dose-depend-ent relation with the exposure of DNF.

关 键 词：二甲基甲酰胺 肝细胞 凋亡相关蛋白 

分 类 号：R575.5[医药卫生—消化系统]