Shimazaki Takahiro, Noro Nobuhiro, Hagikura Kazuhiro, Matsumoto Taro, Yoshida-Noro Chikako
Department of Applied Molecular Chemistry, Graduate School of Industrial Technology, Nihon University, Narashino, Chiba 275-8575, Japan.
GlaxoSmithKline Pharmaceuticals Ltd., Minato-ku, Tokyo 107-0052, Japan.
Biology (Basel). 2021 Nov 20;10(11):1212. doi: 10.3390/biology10111212.
(1) Background: The control of angiogenesis is essential in disease treatment. We investigated angiogenesis-promoting or -suppressing factors and their molecular mechanisms. (2) Methods: Angiogenesis from HUVECs was quantitatively analyzed using the Angiogenesis Analysis Kit (Kurabo, Osaka, Japan). Human rAng-1-producing 107-35 CHO cells or mouse DFAT-D1 cells were co-cultured with HUVEC. Antioxidant polyphenols were added to the culture. Gene expression was analyzed by RT-PCR. (3) Results: The addition of rAng-1-producing cells, their culture supernatant, or commercially available rAng-1 showed a promoting effect on angiogenesis. The co-culture of DFAT-D1 cells promoted angiogenesis. Polyphenols showed a dose-dependent inhibitory effect on angiogenesis. Luteolin and quercetin showed remarkable anti-angiogenic effects. The expression of vWF, Flk1, and PECAM-1 was increased by adding rAng-1-producing cell culture supernatant. Polyphenols suppressed these genes. Apigenin and luteolin markedly suppressed α-SMA and Flk1. Resveratrol and quercetin enhanced the expression of PPARγ, and luteolin suppressed the expression of COX-1. The expression of endothelial nitric oxide synthase (eNOS), an oxidative stress-related gene, was slightly increased by luteolin. These results suggest that polyphenols induce ROS reduction. (4) Conclusions: We showed the promoting effect of Ang-1 or DFAT and the suppressing effect of polyphenols on angiogenesis and studied their molecular mechanisms. These results help control angiogenesis in regenerative therapy.
(1) 背景：血管生成的控制在疾病治疗中至关重要。我们研究了促血管生成或抑血管生成因子及其分子机制。(2) 方法：使用血管生成分析试剂盒（日本大阪库拉博公司）对人脐静脉内皮细胞（HUVECs）的血管生成进行定量分析。将产生人重组血管生成素-1（rAng-1）的107-35 CHO细胞或小鼠去分化脂肪细胞（DFAT-D1）与HUVEC共同培养。向培养物中添加抗氧化多酚。通过逆转录聚合酶链反应（RT-PCR）分析基因表达。(3) 结果：添加产生rAng-1的细胞、其培养上清液或市售rAng-1对血管生成有促进作用。DFAT-D1细胞的共培养促进了血管生成。多酚对血管生成显示出剂量依赖性抑制作用。木犀草素和槲皮素显示出显著的抗血管生成作用。添加产生rAng-1的细胞培养上清液可增加血管性血友病因子（vWF）、胎儿肝激酶1（Flk1）和血小板内皮细胞黏附分子-1（PECAM-1）的表达。多酚抑制这些基因。芹菜素和木犀草素显著抑制α-平滑肌肌动蛋白（α-SMA）和Flk1。白藜芦醇和槲皮素增强过氧化物酶体增殖物激活受体γ（PPARγ）的表达，而木犀草素抑制环氧合酶-1（COX-1）的表达。木犀草素使氧化应激相关基因内皮型一氧化氮合酶（eNOS）的表达略有增加。这些结果表明多酚可诱导活性氧（ROS）减少。(4) 结论：我们展示了血管生成素-1或DFAT对血管生成的促进作用以及多酚对血管生成的抑制作用，并研究了它们的分子机制。这些结果有助于在再生治疗中控制血管生成。
