摘要
目的 :探讨乙醛脱氢酶2(ALDH-2)在内皮祖细胞(EPCs)的氧化应激反应中的调节作用和机制。方法 :培养人外周血EPCs,分别用空白对照、1μmol/L ALDH-2特异性激活剂(Alda-1)、1μmol/L叔丁基过氧化氢(tBHP)和1μmol/L Alda-1预处理+1μmol/L tBHP干预,再分别用2',7'-Dichlorofluorescein diacetate(DCFH-DA)和5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide(JC-1)染色EPCs,检测EPCs的自由氧水平和线粒体膜电位,采用Transwell小室评价EPCs的迁移能力,采用蛋白免疫印迹(Western blot)评价EPCs的p38信号通路激活情况。结果 :tBHP干预和Alda-1预处理+tBHP干预后的自由氧水平相对于空白对照分别为(441.7±24.8)%和(237.4±12.0)%,差异均有统计学意义(P<0.05)。空白对照、tBHP干预和Alda-1预处理+tBHP干预的EPCs发生线粒体膜电位丢失的细胞比率分别为(5.7±2.1)%,(81.7±3.7)%和(37.4±3.2)%;EPCs的迁移细胞数分别为108±9/高倍视野,22±4/高倍视野和67±7/高倍视野,三者间差异均有统计学意义(P<0.05)。加入tBHP后EPCs的p38信号通路增强[信号强度为空白对照的(259.1±7.7)%],激活ALDH-2后,tBHP引起的p38信号活动被减弱[为空白对照的(186.4±8.0)%]。结论 :ALDH-2能够减少EPCs氧化应激反应,减少EPCs线粒体膜损伤,保护EPCs的迁移功能。而这可能与p38信号通路有关。
Objective：To investigate the role of aldehyde dehydrogenase-2（ALDH-2） for regulating human endothelial progenitor cells（EPCs） oxidative stress reaction and its mechanism. Methods： Human EPCs were isolated from peripheral blood of healthy adults and the cells were cultured in 4 groups：（1） Blank control group,（2）Alda-1 group, the cells were treated by 1μmol/L Alda-1, a specific activator of ALDH-2,（3）tB HP（10μg/ml） group and（4）Alda-1 pretreatment＋tBHP group. EPCs reactive oxygen species（ROS） levels were evaluated by DCFH-DA staining, mitochondrial membrane potentials were detected by JC-1 method, migration capacity was measured by transwell chamber method and the activation of p38 signal pathway was examined by Western blot analysis.Results： Compared with Blank control group, ROS levels in tBHP group and Alda-1 pretreatment＋tBHP group were（441.7 ± 24.8） % and（237.4 ± 12.0） %, all P〈0.05. In Blank control group, tBHP group and Alda-1 pretreatment＋tBHP group, the proportion of EPCs lost their mitochondrial membrane potentials were（5.7 ± 2.1） %,（81.7 ± 3.7） % and（37.4 ± 3.2） % respectively, all P〈0.05; the number of EPCs migration were（108 ± 9）/HP,（22 ± 4）/HP and（67 ± 7）/HP respectively,all P〈0.05. Compared with Blank control group, the activation of p38 signal pathway increased to（259.1 ± 7.7） % in tB HP group, while it was reduced to（186.4 ± 8.0） % in Alda-1 pretreatment＋tB HP group.Conclusion： ALDH-2 could reduce ROS level in human EPCs, it may decrease mitochondrial membrane damage, protect migration which might be related to p38 signal pathway.
出处
《中国循环杂志》
CSCD
北大核心
2016年第5期502-507,共6页
Chinese Circulation Journal
基金
湖北省自然科学基金面上资助项目(2014CFC1055)
关键词
内皮祖细胞
乙醛脱氢酶2
氧化应激
线粒体膜电位
Endothelial progenitor cells
Aldehyde dehydrogenase-2
Oxidative stress
Mitochondrial membrane potentials
