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目的探讨在回转模拟失重条件下自噬对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)血管生成能力的影响。方法体外培养HUVECs,随机分为回转组、静止对照组、静止+抑制剂预处理组及回转+抑制剂预处理组,选用的抑制剂为自噬特异性抑制剂3-Methyladenine(3-MA)。Western Blot检测HUVECs中微管相关蛋白轻链3(LC3)和Bcl-2相关蛋白(Beclin1)的表达,划痕实验及Matrigel胶小管生成实验观察各组HUVECs迁移及血管生成能力变化。结果回转24 h后,HUVECs中LC3及Beclin1蛋白的表达均显著高于对照组(P<0.05)。24 h回转使HUVECs的迁移能力及血管生成能力明显增强(P<0.05),且这种增强效应可以被3-MA预处理所抑制。结论回转模拟失重可以诱导HUVECs血管生成能力增强,其发生机制可能与自噬的激活有关。
Objective To investigate the effect of autophagy on the angiogenesis of human umbilical vein endothelial cells (HUVEC) under rotational simulated weightlessness. Methods HUVECs were cultured in vitro and were randomly divided into three groups: the control group, the static control group, the pretreatment group of static inhibitor and the inhibitor of rotation plus inhibitor. The selected inhibitor was 3-Methyladenine (3-MA) . Western Blot was used to detect the expression of LC3 and Beclin1 in HUVECs. Scratch assay and Matrigel tube formation assay were used to observe the migration and angiogenesis ability of HUVECs in each group. Results The expression of LC3 and Beclin1 protein in HUVECs were significantly higher than those in control group (P <0.05) after 24 h of rotation. After 24 h of rotation, the migration ability and angiogenesis ability of HUVECs were significantly enhanced (P <0.05), and this enhancement effect could be inhibited by 3-MA pretreatment. Conclusion Swirling simulated weightlessness can induce angiogenesis in HUVECs. The mechanism may be related to the activation of autophagy.