摘要
目的探讨镉在诱导人胚胎肾细胞(HEK293)凋亡中氧化应激的作用。方法采用流式细胞仪检测细胞凋亡率;采用流式细胞仪和激光共聚焦检测胞浆内的活性氧(ROS)水平,采用分光光度法检测细胞内丙二醛(MDA)、还原型谷胱甘肽(GSH)含量,蛋白印迹法检测Bcl-2蛋白、Caspase-3酶原表达量的变化。结果在一定浓度范围内随着镉浓度的升高,凋亡率升高。在60μmol/L氯化镉诱导细胞6h,凋亡率达到最高;细胞内ROS生成显著增多;MND含量明显增多(P<0.01),GSH含量大幅下降(P<0.01);Bcl-2表达量较镉处理3h时表达量有所下降;Cas-pase-3酶原蛋白表达下降。结论镉诱导HEK293细胞凋亡的机制与ROS上升及其导致的氧化损伤、Bcl-2蛋白表达下调和Caspase-3的激活有关。
Objective To explore the effect of oxidative stress on the apoptosis of HEK293 cells induced by cadmium. Methods The apoptosis were measured by flow cytometry, and the content of cytosolic reactive oxygen species（ROS） were measured respectively by flow cytometry and confocal laser microscopy. The HEK293 cells homogenates were prepared to de- tect the levels of malondialdehyde（MDA） and reduced glutathi0ne hormone （GSH）in spectro photometric assay. Western blot was used to analyze the expressions of Bcl-2 and Caspase-3 proenzyme. Results The rate of apoptosis exhibited in the dose - dependent manner in definite CdC12 concentrations extent. After six hours with treatment 60 μmol/L CdCI2, the rate of apoptosis was in peak, the levels of cytosolic ROS and MDA increased and the level of GSH decreased significantly（ P 〈 0.01 ）. The expression of Bcl-2 was lower compared with three-hour treatment with 60 btmol/L CdC12, and the expression of Caspase-3 proenzyme decreased compared with the control. Conclusion Cd-induced apoptosis involves enhancive ROS, oxidative injury, the expression of Bcl-2 down regulation and the activation of Caspase-3 in HEK293 cells.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2007年第10期1192-1193,共2页
Chinese Journal of Public Health
基金
江苏省教育厅自然科学基金(2005104TSJB157)
关键词
镉
凋亡
氧化应激
cadmium
apotosis
oxidative stress
