内毒素体外诱导人肝细胞凋亡的研究  

THE STUDY OF LIPOPOLYSACCHARIDE-INDUCED APOPTOSIS OF HUMAN HEPATOCYTE IN VITRO

作　　者：吴刚[1] 闫歌[1] 巫国谊[1] 高小林[1] 张建军[1] 张大志[1] 郭树华[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,重庆400010

出　　处：《泸州医学院学报》2004年第4期289-291,共3页Journal of Luzhou Medical College

摘　　要：目的 :探讨内毒素 (LPS)在体外直接作用于LO2细胞 ,引起细胞凋亡所表现出的病理及生物化学特性的变化。方法 :LPS诱导LO2细胞的凋亡。将LO2细胞培养 2 4h后换用含LPS( 2 0 μg/ml)的新鲜培养液 ,设多个时间点 ,提取肝细胞的DNA片段经琼脂糖凝胶电泳观察肝细胞生物化学性质的改变 ;用透射电镜观察其形态学变化 ;TUNEL标记法观察细胞凋亡。结果 :LPS直接作用的LO2细胞在形态学上表现出胞浆浓缩核凝聚成团块、形成凋亡小体、细胞器机构基本正常等凋亡细胞的特征 ;在生物化学方面表现典型的凋亡梯形。肝细胞凋亡在处理后 2 4小时之内随处理时间的延长而增加。TUNEL检测结果显示染色阳性细胞。结论 :LPS在体外的直接作用可诱导正常肝细胞发生凋亡。Objective: To investigate the effect of lipopolysaccharide(LPS) on LO2 cells in vitro. Medthod: The LO2 cells were cultured in vitro for 24h, then the fresh culture medium with LPS(20μg/ml) was added. The cells were collectod for examination after further culture for 3, 8, 16, and 24 hours. DNA-fragment was analyzed by agarose gel electrophoresis to determine the biochemical apoptostic changes. Transmission electron microscopy was used to observe the morphological changes of hepatocyte. The apoptosis of LO2 cells was detected by terminal deoxynucleotide transferas-mediated aUTP nick end labeling(TUNEL). Result: LO2 cells treated with LPS exhibited typical apoptosis-specific morphological features. The DNA-fragments by agarose gel electrophoresis demonstrated the typical ladder pattern and the positive result was observed by TUNEL. The apoptosis increased with the time within 24h when hepatocytes were exposed to LPS alone. Conclusion: LPS can induce apoptosis of hepatocyte in vitro.

关 键 词：内毒素 凋亡 肝细胞 

分 类 号：R512.6[医药卫生—内科学]