摘要
目的探讨氢醌（HQ）对体外培养L-02肝细胞凋亡（apoptosis）的影响和HQ毒性作用的分子机制。方法采用噻唑蓝（MTT）比色法测定不同浓度HQ（0,5,10,20,40,80,160和320μmol/L）作用24 h后L-02肝细胞的相对存活率;采用DNA琼脂糖凝胶电泳和流式细胞术检测HQ染毒后的细胞凋亡状况。结果HQ在0~80μmol/L的染毒剂量范围内,对L-02肝细胞的存活率没有明显的影响（P〉0.05）,当HQ染毒剂量超过160μmol/L时,L-02肝细胞的存活率则明显地下降（P〈0.01）。10,20,40,80,160和320μmol/L组L-02肝细胞DNA琼脂糖凝胶电泳出现凋亡特征性梯状条带,并且随着HQ染毒剂量增加而渐趋明显。流式细胞术检测显示,HQ各染毒剂量组（5,10,20,40,80,160和320μmol/L）作用24 h后均可引起L-02肝细胞的调亡,并呈现出一定的剂量—反应关系;此外,还可出现明显的亚二倍体峰。结论HQ在体外能够诱导L-02肝细胞发生凋亡。
Objective To study the apoptotic changes of L-02 hepatic cells induced by hydroquinone （HQ）in vitro and its molecular mechanism of toxicity. Methods MTT assay was used to detect the effects of HQ with various concentrations（ 0,5,10,20,40,80,160 and 320 μmol/L） on the survival rate of L-02 hepatic cells for 24 h. Apoptosis of L-02 hepatic cells treated with HQ was detected by DNA agarose gel electrophoresis and flow cytometry. Results MTT assay showed that HQ with concentration from 0 to 80 μmol/L has little effect on the viability of L-02（P〉0.05）, while the viabilities in groups of 160 μmol/L and 320 μmol/L were significantly lowered than those in the control（P〈0.01）after treated with HQ for 24 hours. DNA ladder occurred in 10, 20, 40, 80, 160 and 320 μmol/L groups. As the dose of HQ increased, DNA ladder appeared to be more and more obvious. The flow cytornetry showed that the apoptosis rates of L-02 hepatic cells treated with HQ（5, 10,20, 40, 80,160 and 320 μmol/L）for 24 h were 4. 29%, 4.61%, 5.07G, 5.45%, 7.87%, 10.16% and 18.30% respectively, showing significant dose-effect relationship. In addition, a typical sub-diploid peak could distinctively appear. Conclusion Apoptosis of L-02 hepatic cells could be effectively induced by HQ in vitro.
出处
《工业卫生与职业病》
CAS
CSCD
北大核心
2007年第5期274-277,共4页
Industrial Health and Occupational Diseases
基金
国家重点基础研究发展计划(973)项目(2002CB512903
2002CB512904)
