二乙基亚硝胺对人胎肝干细胞的毒性效应  

THE CYTOTOXICITY EFFECT OF DIETHYLNITROSAMINE IN HUMAN HEPATIC STEM CELLS

作　　者：彭哲[1] 杨丹凤[2] 颜培华[2] 肖忠海[2] 李曦[2] 卫华[2] 曹燕卿[2] 张竞丹[2] 

机构地区：[1]广西医科大学研究生院,南宁530121 [2]军事医学科学院卫生学环境医学研究所

出　　处：《解放军预防医学杂志》2012年第3期164-167,共4页Journal of Preventive Medicine of Chinese People's Liberation Army

摘　　要：目的探讨二乙基亚硝胺(Diethylnirtosamine,DENA)对人胎肝干细胞的毒性效应,为评价其遗传毒性提供理论依据。方法用不同质量浓度DENA(0、450、900、1350、1800、2250μg/ml)处理人胎肝干细胞后,应用MTT法检测DENA对体外培养的人胎肝干细胞增殖的抑制作用及量效关系;通过单细胞凝胶电泳实验检测不同剂量DENA处理后人胎肝干细胞DNA的损伤情况;利用Hoechst 33258染色方法检测DENA诱导后人胎肝干细胞凋亡的情况。结果人胎肝干细胞经900、1350、1800μg/ml的DENA处理48 h后,细胞存活率低于阴性对照组,差异有显著性(P<0.01);人胎肝干细胞经DENA处理24 h后,出现染色体固缩、浓染等细胞凋亡表现;单细胞凝胶电泳实验发现,当DENA为900和1350μg/ml时,具有致DNA断裂的作用,尾部DNA含量分别达到(18.44±4.99)%和(17.33±3.29)%,明显高于对照组的(0.015±0.004)%,差异有显著性(P<0.01),但在1800和2250μg/ml时,彗星拖尾程度明显降低,差异有显著性(P<0.01)。结论 DENA对人胎肝干细胞具有直接毒性效应,引起DNA损伤,具有潜在遗传毒性。Objective To investigate the cytotoxieity and DNA damage induced by diethylnirtosamine（DENA） in human hepatic stem cell, so as to provide theoretical basis for evaluating its genetic toxicity. Methods Human hepatic stem cells were exposed to six doses of DENA（0,450,900, 1350, 1800,2250 μg/ml）. Cell survival and DNA damage were evaluated by methyl thiazolyl tetrazolium （MTT） assay and single eell gel eleetrophoresis （SCGE） respectively. Hoechst 33258 staining was used to detect the apoptosis of fetal hepatic stem cells. Results The survival rate of fetal hepatie stem cell was significantly lower than that of control group（ P 〈 0.05） at the dose of 900, 1350, and 1800 μg/ml after 48 h treatment. When hepatic stem cells were exposed to DENA for 24 h, cellular apoptosis occurred with karyopyknosis and deep staining. Break in DNA strands was found at 900, 1350/lg/ml of DENA, the content of DNA in tail [ （ 18.44 ±4.99） %, （ 17.33 ±3.29） % respectively] was higher than that of the control [ （0. 015 ± 0. 004） % ] （ P 〈 0.01 ）, while the comet length was decreased at higher doses（ 1800,2250 μg/ml）（ P 〈 0.01 ）. Conclusion DENA could induce direet DNA damage of human hepatic stem cells in vitro, so it might have some potential genetic toxicity.

关 键 词：人胎肝干细胞 二乙基亚硝胺 细胞毒性 MTT 单细胞凝胶电泳 凋亡 

分 类 号：R994.6[医药卫生—毒理学]