中华预防医学杂志
中華預防醫學雜誌
중화예방의학잡지
CHINESE JOURNAL OF
2009年
1期
56-60
,共5页
胡恭华%庄志雄%黄海燕%庾蕾%杨淋清%纪卫东
鬍恭華%莊誌雄%黃海燕%庾蕾%楊淋清%紀衛東
호공화%장지웅%황해연%유뢰%양림청%기위동
氢醌类%肝细胞%DNA损伤
氫醌類%肝細胞%DNA損傷
경곤류%간세포%DNA손상
Hydroquinones%Hepatocytes%DNA damage
目的 研究氢醌(hydmquinone,HQ)对L-O2人肝细胞中跨损伤合成DNA聚合酶η(Pohη)表达及DNA损伤的影响,探讨Polη在DNA损伤耐受过程中的作用及其可能机制.方法 将L-O2人肝细胞用不同浓度(0、5、10、20、40、80和160 μmol/L) 的HQ作用24 h之后,采用噻唑蓝(MTY)比色法测定细胞相对存活率;单细胞凝胶电泳检测DNA损伤情况;实时荧光定量PCR和Western blotting 技术检测Polη在mRNA 和蛋白质水平上的表达.结果 在0～80 μmol/L 的范围内,HQ对L-O2人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,其存活率则降为(79.20±7.94)%,与对照组(100.00±3.71)%比较,差异有统计学意义(F=17.11,P＜0.01).随着HQ作用浓度的升高,L-O2人肝细胞DNA链的断裂程度也随着逐渐增加.在HQ染毒剂量0～80μmol/L 的范围内,Polη在mRNA水平(相对定量值依次为1.00±0.00、1.20±0.09、2.02±0.19、2.37±0.10、2.67±0.16和4.40±0.18)和蛋白质水平(相对定量值依次为0.22、0.24、0.34、0.44、0.45和1.25)上的表达均有随着剂量的增加而增加的趋势,80 μmol/L 处达到峰值;当HQ的剂量达到160 μmol/L时,Polη的表达则有所降低,mRNA水平和蛋白质水平相对定量值分别为2.32±0.16和1.20,高于对照组.结论 Polη可能参与了HQ所致L-O2 人肝细胞DNA损伤的耐受过程.
目的 研究氫醌(hydmquinone,HQ)對L-O2人肝細胞中跨損傷閤成DNA聚閤酶η(Pohη)錶達及DNA損傷的影響,探討Polη在DNA損傷耐受過程中的作用及其可能機製.方法 將L-O2人肝細胞用不同濃度(0、5、10、20、40、80和160 μmol/L) 的HQ作用24 h之後,採用噻唑藍(MTY)比色法測定細胞相對存活率;單細胞凝膠電泳檢測DNA損傷情況;實時熒光定量PCR和Western blotting 技術檢測Polη在mRNA 和蛋白質水平上的錶達.結果 在0～80 μmol/L 的範圍內,HQ對L-O2人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,其存活率則降為(79.20±7.94)%,與對照組(100.00±3.71)%比較,差異有統計學意義(F=17.11,P＜0.01).隨著HQ作用濃度的升高,L-O2人肝細胞DNA鏈的斷裂程度也隨著逐漸增加.在HQ染毒劑量0～80μmol/L 的範圍內,Polη在mRNA水平(相對定量值依次為1.00±0.00、1.20±0.09、2.02±0.19、2.37±0.10、2.67±0.16和4.40±0.18)和蛋白質水平(相對定量值依次為0.22、0.24、0.34、0.44、0.45和1.25)上的錶達均有隨著劑量的增加而增加的趨勢,80 μmol/L 處達到峰值;噹HQ的劑量達到160 μmol/L時,Polη的錶達則有所降低,mRNA水平和蛋白質水平相對定量值分彆為2.32±0.16和1.20,高于對照組.結論 Polη可能參與瞭HQ所緻L-O2 人肝細胞DNA損傷的耐受過程.
목적 연구경곤(hydmquinone,HQ)대L-O2인간세포중과손상합성DNA취합매η(Pohη)표체급DNA손상적영향,탐토Polη재DNA손상내수과정중적작용급기가능궤제.방법 장L-O2인간세포용불동농도(0、5、10、20、40、80화160 μmol/L) 적HQ작용24 h지후,채용새서람(MTY)비색법측정세포상대존활솔;단세포응효전영검측DNA손상정황;실시형광정량PCR화Western blotting 기술검측Polη재mRNA 화단백질수평상적표체.결과 재0～80 μmol/L 적범위내,HQ대L-O2인간세포적존활솔몰유명현적영향;당염독제량초과160 μmol/L 시,기존활솔칙강위(79.20±7.94)%,여대조조(100.00±3.71)%비교,차이유통계학의의(F=17.11,P＜0.01).수착HQ작용농도적승고,L-O2인간세포DNA련적단렬정도야수착축점증가.재HQ염독제량0～80μmol/L 적범위내,Polη재mRNA수평(상대정량치의차위1.00±0.00、1.20±0.09、2.02±0.19、2.37±0.10、2.67±0.16화4.40±0.18)화단백질수평(상대정량치의차위0.22、0.24、0.34、0.44、0.45화1.25)상적표체균유수착제량적증가이증가적추세,80 μmol/L 처체도봉치;당HQ적제량체도160 μmol/L시,Polη적표체칙유소강저,mRNA수평화단백질수평상대정량치분별위2.32±0.16화1.20,고우대조조.결론 Polη가능삼여료HQ소치L-O2 인간세포DNA손상적내수과정.
Objective To investigate the effects of hydroquinone (HQ) on expression of Polymerase eta (Polη) and DNA damage in human hepatic cells (L-O2),and to explore the role and possible mechanism of Polη involved in the process of DNA damage-tolerance.Methods After L-O2 hepatic cells were exposed to HQ with various concentrations (0,5,10,20,40,80 and 160 μmol/L) for 24 h,cell survival rate was detected by MTT assay;DNA impairment was detected by single cell gel electrophoresis (SCGE);Real-time fluorescent quantitative PCR and Western blotting methods were used to measure the expression of Polη at the mRNA and protein level in L-O2 hepatic cells exposed to HQ with various concentrations(0,5,10,20,40,80 and 160 μmol/L).Resuits MTT assay showed that HQ with concentrations from 0 to 80 μmol/L had little effect on the survival rate of L-O2(P＞0.05);whereas the survival rate of the group of 160 μmol/Lwas significantly higher than that of the control(P＜0.01) after being treated with HQ for 24 h;the higher dose of HQ presented,the more degrees of DNA damage were produced.It was found that HQ in a low concentration (1-80 μmol/L) could induce t11e expression of Polη which was in proprotion to the increasement of HQ concentration;the expression levels of mRNA and protein were reached to the maximum when treated with 80 μmol/L;the expression of Polη decreased (the relative quantity values were 2.32±0.16 and 1.20 respectively) once the concentration of HQ exceeded 160 μmol/L as compared with the group of 80 μmol/L,but it was higher than that of the control.Conclusion This study suggested that Polη might involve in the process of DNA damage-tolerance induced by HQ in the hepatic cells.
