Trenti Annalisa, Tedesco Serena, Boscaro Carlotta, Ferri Nicola, Cignarella Andrea, Trevisi Lucia, Bolego Chiara
Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy
J Pharmacol Exp Ther. 2017 Jun;361(3):398-407. doi: 10.1124/jpet.116.238212. Epub 2017 Mar 27.
The endogenous estrogen 17-estradiol (E2) is a key factor in promoting endothelial healing and angiogenesis. Recently, proangiogenic signals including vascular endothelial growth factor and others have been shown to converge in endothelial cell metabolism. Because inhibition of the glycolytic enzyme activator phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) reduces pathologic angiogenesis and estrogen receptor (ER) signaling stimulates glucose uptake and glycolysis by inducing PFKFB3 in breast cancer, we hypothesized that E2 triggers angiogenesis in endothelial cells via rapid ER signaling that requires PFKFB3 as a downstream effector. We report that treatment with the selective G protein-coupled estrogen receptor (GPER1) agonist G-1 (10 to 10 M) mimicked the chemotactic and proangiogenic effect of E2 as measured in a number of short-term angiogenesis assays in human umbilical vein endothelial cells (HUVECs); in addition, E2 treatment upregulated PFKFB3 expression in a time- and concentration-dependent manner. Such an effect peaked at 3 hours and was also induced by G-1 and abolished by pretreatment with the GPER1 antagonist G-15 or GPER1 siRNA, consistent with engagement of membrane ER. Experiments with the PFKFB3 inhibitor 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one showed that PFKFB3 activity was required for estrogen-mediated HUVEC migration via GPER1. In conclusion, E2-induced angiogenesis was mediated at least in part by the membrane GPER1 and required upregulation of the glycolytic activator PFKFB3 in HUVECs. These findings unravel a previously unrecognized mechanism of estrogen-dependent endocrine-metabolic crosstalk in HUVECs and may have implications in angiogenesis occurring in ischemic or hypoxic tissues.
内源性雌激素17-β-雌二醇（E2）是促进内皮愈合和血管生成的关键因素。最近，包括血管内皮生长因子等在内的促血管生成信号已被证明在内皮细胞代谢中汇聚。由于糖酵解酶激活剂磷酸果糖激酶-2/果糖-2,6-二磷酸酶3（PFKFB3）的抑制可减少病理性血管生成，且雌激素受体（ER）信号通过在乳腺癌中诱导PFKFB3刺激葡萄糖摄取和糖酵解，我们推测E2通过快速ER信号在内皮细胞中触发血管生成，该信号需要PFKFB3作为下游效应器。我们报告，在人脐静脉内皮细胞（HUVECs）的一些短期血管生成试验中，用选择性G蛋白偶联雌激素受体（GPER1）激动剂G-1（10至10 μM）处理模拟了E2的趋化和促血管生成作用；此外，E2处理以时间和浓度依赖性方式上调PFKFB3表达。这种效应在3小时达到峰值，也可由G-1诱导，并被GPER1拮抗剂G-15或GPER1 siRNA预处理消除，这与膜ER的参与一致。使用PFKFB3抑制剂3-(3-吡啶基)-1-(4-吡啶基)-2-丙烯-1-酮的实验表明，PFKFB3活性是雌激素通过GPER1介导的HUVEC迁移所必需的。总之，E2诱导的血管生成至少部分由膜GPER1介导，并且需要上调HUVECs中的糖酵解激活剂PFKFB3。这些发现揭示了HUVECs中雌激素依赖性内分泌-代谢串扰的一种先前未被认识的机制，并且可能对缺血或缺氧组织中发生的血管生成有影响。
