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目的 观察回转器模拟失重对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC-C)血管生成能力的影响,并分析探讨可能涉及的关键信号分子内皮型一氧化氮合成酶(endothelial nitricoxide synthetase,eNOS)的作用。方法体外培养HUVEC-C,随机分为回转模拟失重组、1 G静止对照组及回转+抑制剂组,选用内皮型一氧化氮合成酶的特异性抑制剂L-NAME(N-nitro-L-arginine methyl es-ter hydrochloride)。Matrigel胶小管形成实验观察不同处理组HUVEC-C血管生成能力的变化情况。RT-PCR和Western blot分别检测模拟失重前、后HUVEC-C中eNOS mRNA和蛋白的表达变化。结果24 h模拟失重使HUVEC-C的血管生成能力较对照组明显增强(P<0.01),且这种增强效应可以被L-NAME所抑制。模拟失重24 h后,HUVEC-C中eNOS mRNA和蛋白的表达均显著高于对照组(P<0.05)。结论回转模拟失重可以诱导HUVEC-C血管生成能力增强,其发生机制与eNOS表达上调有关。
Objective To observe the effect of revolver simulated weightlessness on the angiogenesis of human umbilical vein endothelial cells (HUVEC-C) and to explore the possible molecular mechanisms involved in the pathogenesis of endothelial nitric oxide synthetase eNOS) role. Methods HUVEC-C cells were cultured in vitro and were randomly divided into three groups: swirled simulated weightlessness group, 1 G static control group and swivel + inhibitor group. L-NAME (N-nitro-L- arginine methyl es-ter hydrochloride). Matrigel tracheal formation assay was used to observe the changes of HUVEC-C angiogenesis in different treatment groups. RT-PCR and Western blot were used to detect the expression of eNOS mRNA and protein in HUVEC-C before and after simulated weightlessness respectively. Results 24-hour simulated weightlessness significantly enhanced the angiogenic ability of HUVEC-C compared with the control group (P <0.01), and this enhancement effect was inhibited by L-NAME. After simulated weightlessness for 24 h, the expression of eNOS mRNA and protein in HUVEC-C was significantly higher than that in the control group (P <0.05). Conclusion Swirling simulated weightlessness can induce HUVEC-C angiogenesis and its mechanism is related to the up-regulation of eNOS expression.