中华劳动卫生职业病杂志
中華勞動衛生職業病雜誌
중화노동위생직업병잡지
CHINESE JOURNAL OF INDUSTRIAL HYGIENE AND OCCUPATIONAL DISEASES
2009年
4期
222-225
,共4页
胡恭华%庄志雄%黄海燕%庾蕾%袁建辉%杨淋清
鬍恭華%莊誌雄%黃海燕%庾蕾%袁建輝%楊淋清
호공화%장지웅%황해연%유뢰%원건휘%양림청
氢醌类%肝细胞%连接酶类%DNA损伤
氫醌類%肝細胞%連接酶類%DNA損傷
경곤류%간세포%련접매류%DNA손상
Hydroquinone%Hepatoeytes%Ligases%DNA damage
目的 研究氧醌(hydroquinone,HQ)对人L-02肝细胞中泛素连接酶Pad18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160 μmol/L)的HQ作用24 h,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Pad18在蛋白质水平上的表达.结果 在0-80 μmoFL的范嗣内,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160 μmol/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=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40 μmol/L时,Pad18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.
目的 研究氧醌(hydroquinone,HQ)對人L-02肝細胞中汎素連接酶Pad18錶達的影響,探討Rad18在HQ所緻肝細胞毒性中的作用及其可能機製.方法 將L-02肝細胞用不同濃度(0、5、10、20、40、80和160 μmol/L)的HQ作用24 h,採用噻唑藍(MTT)比色法測定細胞存活率;用單細胞凝膠電泳檢測DNA損傷情況;實時熒光定量PCR技術檢測Rad18在mRNA水平上的錶達;Western blot方法檢測Pad18在蛋白質水平上的錶達.結果 在0-80 μmoFL的範嗣內,HQ對L-02肝細胞的存活率沒有明顯的影響,噹染毒劑量超過160 μmol/L時,細胞存活率(79.20%)明顯下降,與對照組比較,差異有統計學意義(P<0.01).隨著 HQ作用濃度的升高,L-02肝細胞的Olive尾矩也逐漸增加,呈線性正相關關繫(r=920,P<0.01).在HQ染毒劑量低于40μmol/L的範圍內,Rad18在mRNA和蛋白質水平上的錶達均有隨著劑量的增加而增加的趨勢;噹HQ的劑量超過40 μmol/L時,Pad18在mRNA水平上的錶達繼續增加,而其在蛋白質水平上的錶達則無明顯變化.結論 HQ可使L-02肝細胞的Rad18錶達增加.
목적 연구양곤(hydroquinone,HQ)대인L-02간세포중범소련접매Pad18표체적영향,탐토Rad18재HQ소치간세포독성중적작용급기가능궤제.방법 장L-02간세포용불동농도(0、5、10、20、40、80화160 μmol/L)적HQ작용24 h,채용새서람(MTT)비색법측정세포존활솔;용단세포응효전영검측DNA손상정황;실시형광정량PCR기술검측Rad18재mRNA수평상적표체;Western blot방법검측Pad18재단백질수평상적표체.결과 재0-80 μmoFL적범사내,HQ대L-02간세포적존활솔몰유명현적영향,당염독제량초과160 μmol/L시,세포존활솔(79.20%)명현하강,여대조조비교,차이유통계학의의(P<0.01).수착 HQ작용농도적승고,L-02간세포적Olive미구야축점증가,정선성정상관관계(r=920,P<0.01).재HQ염독제량저우40μmol/L적범위내,Rad18재mRNA화단백질수평상적표체균유수착제량적증가이증가적추세;당HQ적제량초과40 μmol/L시,Pad18재mRNA수평상적표체계속증가,이기재단백질수평상적표체칙무명현변화.결론 HQ가사L-02간세포적Rad18표체증가.
Objective To investigate the effects of hydroquinone(HQ) on expression of ubiquitin-lig-ating enzyme Rad18 in human hepatic cells (L-02) ,and to explore the role and possible mechanism of Rad18 involved in toxicity of HQ to hepatic cells. Methods After L-02 hepatic cells were exposed to HQ with vari-ous concentrations(0,5,10,20,40,80 and 160 μmol/L) for 24 h,cell survival rate was measured by MTT as-say; DNA impairment was evaluated by single cell gel electrophoresis (SCGE) ; The expression levels of Rad18 mRNA and protein were detected by Real-time fluorescent quantitative polymerase 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 signifi-cantly 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-depen-dent relationship was shown. HQ in a low concentration(0～40 μmol/L) could induce increase in the expression of Rad18 mRNA and protein which was in proprotion to the increasement of HQ concentration;the expression of Rad18 mRNA was enhanced unceasingly,while the expression of Radl8 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 Rad18 mRNA (r=0.919,P<0.01). Conclusion HQ could regulate up the expression of Rad18 in L-02 hepatic cells.
