南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (03): 313-.

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Low shear stress induces human vascular endothelial cell apoptosis by activating Akt signal and increasing reactive oxygen species

张俊霞，王志梅，左广峰，李冰，张俊杰，田乃亮，陈绍良   

  • 出版日期:2013-03-20 发布日期:2013-03-20

低剪切力通过激活Akt 信号和增加活性氧簇诱导人血管内皮细胞凋亡

  • Online:2013-03-20 Published:2013-03-20

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摘要： 目的探讨低剪切力（LSS）对人血管内皮细胞（ECs）凋亡的影响。方法平行板流室对体外培养的ECs施加2 dyne/cm2
LSS 120 min，观察施加LSS前后ECs凋亡及活性氧簇的变化，并观察施加LSS后不同时间点（0，5，15，30，60，120 min）Akt激酶
活性变化。细胞凋亡采用形态学变化、DAPI染色和TUNEL方法检测。细胞内活性氧簇采用dihydroethidium 和mitoSOX检
测。Western blot 检测Akt 激酶活性。结果施加2 dyne/cm2 LSS后120 min，ECs 脱落并出现凋亡。施加LSS后不同时间点
Akt Ser473和Thr308位点活性增加，2 dyne/cm2 LSS作用120 min后ECs线粒体及胞浆水平活性氧簇产生增加。结论Akt活化
与活性氧簇积累参与LSS诱导的EC凋亡。

Abstract: Objective To investigate the effect of low shear stress (LSS) on apoptosis of human vascular endothelial cells (ECs)
and explore the possible mechanisms. Methods Parallel flow chamber was used to apply LSS at 2 dyne/cm2 on EA.hy926 cells
derived from human umbilical vein endothelial cells for 120 min. Cell apoptosis following LSS was verified by morphological
observation, DAPI staining and TUNEL test. The level of intracellular reactive oxygen species (ROS) was measured by
dihydroethidium (DHE) and mitoSOX. Western blotting was performed to detect the activity of Akt in the cells. Results EC
detachment and apoptosis were observed after exposure to 2 dyne/cm2 LSS for 120 min. Time course study showed that the
phosphorylation level of Akt on residues Ser473 and Thr308 was elevated after flow initiation. Exposure to LSS at 2 dyne/cm2
for 120 min resulted in increased ROS production in the ECs at both the mitochondrial and cytoplasmic levels. Conclusion Akt
activation and increased ROS levels are involved in LSS-induced EC apoptosis.