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体外气道上皮细胞系中血管内皮生长因子mRNA和蛋白的表达

Vascular endothelial growth factor mRNA and protein expression in airway epithelial cell lines in vitro.

作者信息

Koyama S, Sato E, Tsukadaira A, Haniuda M, Numanami H, Kurai M, Nagai S, Izumi T

机构信息

Pulmonary Section, The National Chuushin Matsumoto Hospital, Toyooka, Matsumoto, Japan.

出版信息

Eur Respir J. 2002 Dec;20(6):1449-56. doi: 10.1183/09031936.02.00089802.

Abstract

Vascular endothelial growth factor (VEGF) plays multifunctional roles in vascular permeability, repair and remodelling processes, in addition to the maintenance of vascular structure and function. In the present study, the potential of airway epithelial cell lines, BEAS-2B cells and A549 cells, to release and express VEGF in unstimulated and stimulated conditions was evaluated. The secretion and expression of VEGF were evaluated by enzyme-linked immunosorbant assay and by reverse transcriptase-polymerase chain reaction. The isoforms of released VEGF were determined by high-performance liquid chromatography. BEAS-2B cells and A549 cells released VEGF constitutively. Interleukin (IL)-1beta and tumour necrosis factor (TNF)-alpha augmented the release of VEGF in a time- and dose-dependent manner. The released VEGF was 165 amino acid residues in either condition. Pseudomonas aeruginosa lipopolysaccharide (LPS), interferon (IFN)-gamma, smoke extract (SE), neutrophil elastase (NE), and bradykinin stimulated the release of VEGF. Keracinocyte growth factor (KGF), which reduces vascular permeability, also stimulated both cells to release VEGF. VEGF messenger ribonucleic acid (mRNA) was expressed both time- and dose-dependently at 2 h, and declined after 2 h in response to IL-1beta and TNF-alpha. The expression of VEGF mRNA in airway epithelial cells was also augmented by LPS, IFN-gamma, SE, NE, and KGF stimulation. These data suggest that airway epithelial cells may regulate the maintenance of vascular structure and function, as well as vascular permeability, repair and remodelling processes, in a variety of lung conditions by expressing vascular endothelial growth factor.

摘要

血管内皮生长因子(VEGF)除了维持血管结构和功能外,还在血管通透性、修复和重塑过程中发挥多种功能。在本研究中,评估了气道上皮细胞系BEAS - 2B细胞和A549细胞在未刺激和刺激条件下释放和表达VEGF的潜力。通过酶联免疫吸附测定和逆转录 - 聚合酶链反应评估VEGF的分泌和表达。通过高效液相色谱法测定释放的VEGF的异构体。BEAS - 2B细胞和A549细胞组成性地释放VEGF。白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α以时间和剂量依赖性方式增加VEGF的释放。在两种情况下,释放的VEGF均为165个氨基酸残基。铜绿假单胞菌脂多糖(LPS)、干扰素(IFN)-γ、烟雾提取物(SE)、中性粒细胞弹性蛋白酶(NE)和缓激肽刺激VEGF的释放。减少血管通透性的角质形成细胞生长因子(KGF)也刺激这两种细胞释放VEGF。VEGF信使核糖核酸(mRNA)在2小时时呈时间和剂量依赖性表达,并在2小时后因IL - 1β和TNF - α而下降。LPS、IFN - γ、SE、NE和KGF刺激也增强了气道上皮细胞中VEGF mRNA的表达。这些数据表明,气道上皮细胞可能通过表达血管内皮生长因子在多种肺部疾病中调节血管结构和功能的维持以及血管通透性、修复和重塑过程。

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