摘要
【目的】研究过氧化氢(H_2O_2)对EA.hy926内皮细胞锌指蛋白580(zinc finger 580,ZNF580)表达的影响。【方法】先以不同浓度H_2O_2作用内皮细胞EA.hy926,测定细胞上清中乳酸脱氢酶(lactic dehydrogenase,LDH)确定细胞的损伤程度。选择适宜的H_2O_2浓度作用细胞,在不同浓度和不同时间点提取总RNA和蛋白,以RT-PCR检测ZNF580的m RNA表达水平,同时western blotting检测其蛋白表达差异。【结果】随着H_2O_2浓度的升高,LDH释放随之升高,不同浓度H_2O_2作用EA.hy926内皮细胞后,ZNF580在m RNA转录水平和蛋白水平的表达均有增加,并呈现浓度依赖的趋势。同样的浓度为100μmol/L的H_2O_2作用EA.hy926内皮细胞不同时间后,检测可知ZNF580 m RNA和蛋白水平的表达增加,与对照组差异有统计学意义(P<0.05)。【结论】一定浓度和时间的外源性H_2O_2作用内皮细胞可促进ZNF580在一定程度表达上调。
[Objective] To study the effect of hydrogen peroxide （H2O2） on the expression of ZNF580 in endothelial cells （EA. hy926 cells）. [Methods] EA. hy926 cells were treated with different coneentrations of H2O2. The cell-damage degree was identified by detecting the LDH content of cell supernatant. Then the cells were treated with the suitable concentration of H2O2 The total RNA and protein were extraeted in different coneentrations and at different time points. The expression levels of mRNA and protein of ZNF580 were detected by RT-PCR and western blotting respectively. [ Results ] The release of LDH was correlated with the eoneentration of H2O2. After EA. hy926 cells were treated with different concentrations of H2O2, the mRNA and protein levels of ZNF580 all inereased and showed a concentration-related tendency. After the cells were treated with 100 μmol/L H2O2, the mRNA and protein levels of ZNF580 inereased at different time points and showed signifieant difference compared with control group （P〈 0.05）. [Conclusion] ZNF580 is upregulated by exogenous H202 in certain concentration and at certain time point.
作者
任党利
王麒
黄磊
刘冀琴
REN Dang-li WANG Qi HUANG Lei LIU Ji-qin(Department of Clinical Laboratory, Affiliated Hospital of Logistics University of PAP, Tianjin 300162, China)
出处
《武警后勤学院学报（医学版）》
CAS
2016年第11期869-872,共4页
Journal of Logistics University of PAP（Medical Sciences）
