氢醌对人L-02肝细胞泛素连接酶Rad18表达的影响  

Effect of hydroquinone on expression of ubiquitin-ligating enzyme Radl8 in human L-02 hepatic cells

作　　者：胡恭华[1] 庄志雄[2] 黄海燕[2] 庾蕾[2] 袁建辉[3] 杨淋清[2] 

机构地区：[1]赣南医学院预防医学系,赣州341000 [2]深圳市疾病预防控制中心毒理研究室 [3]中山大学公共卫生学院

出　　处：《中华劳动卫生职业病杂志》2009年第4期222-225,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家重点基础研究发展计划（“973”计划）项目（2002CB512903,2002CB512904）

摘　　要：目的研究氢醌（hydroquinone，HQ）对人L-02肝细胞中泛素连接酶Rad18表达的影响，探讨Rad18在HQ所致肝细胞毒性中的作用及其可能机制。方法将L-02肝细胞用不同浓度（0、5、10、20、40、80和160μmol／L）的HQ作用24h，采用噻唑蓝（MTT）比色法测定细胞存活率；用单细胞凝胶电泳检测DNA损伤情况；实时荧光定量PCR技术检测Rad18在mRNA水平上的表达；Western blot方法检测Rad18在蛋白质水平上的表达。结果存0～80μmol／L的范同内，HQ对L-02肝细胞的存活率没有明显的影响，当染毒剂量超过160μmol／L时，细胞存活率（79．20％）明显下降，与对照组比较，差异有统计学意义（P〈0．01）.随着HQ作用浓度的升高，L-02肝细胞的Olive尾矩也逐渐增加，呈线性正相关关系（r=0．920，P〈0．01）。在HQ染毒剂量低于40μmol／L的范围内，Rad18在mRNA和蛋白质水平上的表达均有随着剂量的增加而增加的趋势；当HQ的剂量超过40μmol／L时，Rad18在mRNA水平上的表达继续增加，而其在蛋白质水平上的表达则无明显变化。结论HQ可使L-02肝细胞的Rad18表达增加。Objective To investigate the effects of hydroquinone（HQ ） on expression of ubiquitin-ligating enzyme Radl 8 in human hepatic cells （L-02）, and to explore the role and possible mechanism of Rad 18 involved in toxicity of HQ to hepatic cells. Methods After L-02 hepatic cells were exposed to HQ with various concentrations （0,5,10,20,40,80 and 160 μmol/L） for 24 b, cell survival rate was measured by MTT assay ;DNA impairment was evaluated by single cell gel electrophoresis （SCGE） ;The expression levels of Rad 18 mRNA and protein were detected by Real-time fluorescent quantitative polymcrase chain reaction （QPCR） technique and Western blot method respectively. Results HQ with concentration from 0 to 80 μmol/L had little effect on survival rate of L-02（P〉0.05 ）;Whereas the survival rate in the group of 160 μmol/L was significantly lower than in the control with the significant difference （P〈0.01） after treated with HQ for 24 h ;The higher dose of HQ presented, the more degrees of olive tail moment （OTM） were produced and a dose-dependent relationship was shown. HQ in a low concentration （0-40 μmol/L ） could induce increase in the expression of Radl8 mRNA and protein which was in proprotion to the increasement of HQ c, oncentration;the expression of Rad18 mRNA was enhanced unceasingly,while the expression of Rad18 protein unchanged basically once the concentration of HQ exceeded 40 μmol/L ; Besides, there was a positive correlation between OTM and the expression level of Radl 8 mRNA （r=0.919, P〈0.01 ）. Conclusion HQ could regulate up the expression of Radl 8 in L-02 hepatic cells.

关 键 词：氢醌类 肝细胞 连接酶类 DNA损伤 

分 类 号：R686[医药卫生—骨科学]