Luteolin Inhibits Smooth Muscle Cell Migration and Proliferation by Attenuating the Production of Nox4, P-Akt and VEGF in Endothelial Cells
Luteolin Inhibits Smooth Muscle Cell Migration and Proliferation by Attenuating the Production of Nox4, P-Akt and VEGF in Endothelial Cells
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DOI：
10.2174/1389201015666140113113843
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发表时间：
2013-01-01
影响因子：
2.8
通讯作者：
Wang, Cheng
中科院分区：
文献类型：
--
作者：
Ding, Hong;Li, Dongye;Wang, Cheng
目的：本研究旨在观察毛地黄黄酮对血管紧张素Ⅱ（Ang Ⅱ）诱导的内皮细胞（EC）损伤的保护作用，探讨血管内皮功能障碍在血管平滑肌细胞增殖和迁移中的作用，并初步探讨其作用机制。研究方法：使用transwell系统建立非接触共培养系统; EC在下部威尔斯孔中培养，而平滑肌细胞（SMC）在上部威尔斯孔中培养。MTT法检测细胞增殖情况。观察并计数通过transwell系统膜的SMCs数量。Western blotting检测内皮细胞VEGF、p-Akt、Nox 4蛋白的表达水平。逆转录-聚合酶链反应（RT-PCR）检测VEGF mRNA的表达。采用酶联免疫吸附法（ELISA）检测内皮细胞培养上清中VEGF的含量。结果如下：Ang Ⅱ刺激的ECs可显著促进SMCs的增殖和迁移，毛地黄黄酮预处理可抑制这种作用。Luteolin可抑制Ang II诱导的内皮细胞Nox 4、p-AKT和VEGF表达的上调。结论：这些结果表明，毛地黄黄酮能够通过抑制Nox 4、p-Akt和VEGF的上调来抑制Ang II诱导的内皮功能障碍，从而抑制由损伤的EC诱导的SMC的增殖和迁移。
Objective: The goals of this study were to observe how luteolin protects endothelial cells (ECs) from injury stimulated by Angiotensin II (Ang II), investigate the role of vascular endothelial dysfunction in vascular smooth muscle cell proliferation and migration in vitro and investigate its primary mechanism of action. Methods: A non-contact co-culture system was established using a transwell system; ECs were cultured in the lower wells, while the smooth muscle cells (SMCs) were cultured in the upper wells. Cell proliferation was assessed by the MTT assay. The number of SMCs that migrated through the membrane of transwell system were observed and counted. The expression levels of various proteins (VEGF, p-Akt, Nox4) expressed in ECs were determined by Western blotting. VEGF mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). The supernatants of ECs were measured by enzyme linked immunosorbent assay (ELISA) to assay VEGF concentration. Results: Ang II-stimulated ECs significantly increased the proliferation and migration of SMCs, and these effects were inhibited by luteolin pretreatment. Luteolin suppressed the Ang II-induced upregulation of Nox4, p-AKT and VEGF expression in ECs. Conclusion: These results demonstrate that luteolin is capable of inhibiting endothelial dysfunction induced by Ang II by suppressing the upregulation of Nox4, p-Akt and VEGF, thereby restraining the proliferation and migration of SMCs induced by injured ECs.