中国组织工程研究Issue(1)：67-71,5.DOI:10.3969/j.issn.2095-4344.2015.01.012
自噬激活如何影响大鼠内皮祖细胞凋亡、增殖和周期的变化?
How does autophagy activation affect the apoptosis, proliferation and cycle of endothelial progenitor cells in rats?
摘要
Abstract
BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.关键词
干细胞/培养/雷帕霉素/内皮祖细胞/LC3-Ⅱ蛋白/自噬/细胞凋亡/细胞增殖/细胞周期/国家自然科学基金Key words
Sirolimus/Autophagy/Cell cycle/Apoptosis/Cell Proliferation分类
医药卫生引用本文复制引用
刘辉,李晓强,朱人大,孟庆友,卢辉俊..自噬激活如何影响大鼠内皮祖细胞凋亡、增殖和周期的变化?[J].中国组织工程研究,2015,(1):67-71,5.基金项目
国家自然科学基金(30972941)@@@@the National Natural Science Foundation of China, No.30972941 （30972941）
