Lucchesi Martina, Di Marsico Lorenza, Guidotti Lorenzo, Lulli Matteo, Filippi Luca, Marracci Silvia, Dal Monte Massimo
Department of Biology, University of Pisa, 56126 Pisa, Italy.
Department of Experimental and Clinical Biomedical Sciences "Mario Serio", University of Florence, 50121 Florence, Italy.
Int J Mol Sci. 2025 Apr 24;26(9):4043. doi: 10.3390/ijms26094043.
β-adrenoceptors (BARs) are involved in vascular endothelial growth factor (VEGF) production during retinal neovascularization. Here, using human retinal endothelial and Müller cells (hRECs and MIO-M1, respectively), we evaluated the effects exerted by hypoxia on BARs, hypoxia-inducible factor-1α subunit (HIF-1α) and VEGF, as well as the involvement of BAR3 and nitric oxide synthase (NOS) enzymes in hypoxia-induced VEGF production. We altered oxygen availability through a hypoxic incubator. BARs, HIF-1 α and VEGF levels were evaluated. Cells were treated with the BAR3 antagonist SR59230A, different NOS inhibitors or the NO donor SNAP. The influence of the BAR3/NOS axis on hypoxic VEGF production was assessed. Hypoxia upregulated BAR3, HIF-1α and VEGF in hRECs and MIO-M1 cells. SR59230A counteracted hypoxia-dependent VEGF increase in both cell lines, exerting no effect on HIF-1α upregulation. Treatments with NOS inhibitors prevented the hypoxia-dependent VEGF increase, while SNAP abrogated the effect of SR59230A in reducing hypoxia-induced VEGF upregulation. The present results corroborate the hypothesis that in the hypoxic retina, BAR3 influence on VEGF production is mediated by NO and suggest that, at least in endothelial and Müller cells, BAR3 activity is necessary to allow the HIF-1-mediated VEGF upregulation.
β-肾上腺素能受体（BARs）参与视网膜新生血管形成过程中血管内皮生长因子（VEGF）的产生。在此，我们分别使用人视网膜内皮细胞和 Müller 细胞（分别为 hRECs 和 MIO-M1），评估了缺氧对 BARs、缺氧诱导因子-1α亚基（HIF-1α）和 VEGF 的影响，以及 BAR3 和一氧化氮合酶（NOS）酶在缺氧诱导的 VEGF 产生中的作用。我们通过缺氧培养箱改变氧的可利用性。评估了 BARs、HIF-1α和 VEGF 的水平。用 BAR3 拮抗剂 SR59230A、不同的 NOS 抑制剂或 NO 供体 SNAP 处理细胞。评估了 BAR3/NOS 轴对缺氧诱导的 VEGF 产生的影响。缺氧上调了 hRECs 和 MIO-M1 细胞中的 BAR3、HIF-1α和 VEGF。SR59230A 抵消了两种细胞系中缺氧依赖性的 VEGF 增加，对 HIF-1α的上调没有影响。用 NOS 抑制剂处理可防止缺氧依赖性的 VEGF 增加，而 SNAP 消除了 SR59230A 在降低缺氧诱导的 VEGF 上调方面的作用。目前的结果证实了以下假设：在缺氧的视网膜中，BAR3 对 VEGF 产生的影响是由 NO 介导的，并且表明，至少在内皮细胞和 Müller 细胞中，BAR3 的活性对于允许 HIF-1 介导的 VEGF 上调是必要的。
