Boscaro Carlotta, Trenti Annalisa, Baggio Chiara, Scapin Chiara, Trevisi Lucia, Cignarella Andrea, Bolego Chiara
Department of Pharmaceutical and Pharmacological Sciences, University of Padova, Padova, Italy.
Department of Medicine, University of Padova, Padova, Italy.
Front Pharmacol. 2020 Dec 17;11:587221. doi: 10.3389/fphar.2020.587221. eCollection 2020.
Female hormones and sex-specific factors are established determinants of endothelial function, yet their relative contribution to human endothelium phenotypes has not been defined. Using human umbilical vein endothelial cells (HUVECs) genotyped by donor's sex, we investigated the influence of sex and estrogenic agents on the main steps of the angiogenic process and on key proteins governing HUVEC metabolism and migratory properties. HUVECs from female donors (fHUVECs) showed increased viability ( < 0.01) and growth rate ( < 0.01) compared with those from males (mHUVECs). Despite higher levels of G-protein coupled estrogen receptor (GPER) in fHUVECs ( < 0.001), treatment with 17β-estradiol (E2) and the selective GPER agonist G1 (both 1-100 nM) did not affect HUVEC viability. Migration and tubularization under physiological conditions were higher in fHUVECs than in mHUVECs ( < 0.05). E2 treatment (1-100 nM) upregulated the glycolytic activator PFKFB3 with higher potency in fHUVECs than in mHUVECs, despite comparable baseline levels. Moreover, Y576/577 phosphorylation of focal adhesion kinase (FAK) was markedly enhanced in fHUVECs ( < 0.001), despite comparable Src activation levels. While the PI3K inhibitor LY294002 (25 µM) inhibited HUVEC migration ( < 0.05), Akt phosphorylation levels in fHUVECs and mHUVECs were comparable. Finally, digitoxin treatment, which inhibits Y576/577 FAK phosphorylation, abolished sexual dimorphism in HUVEC migration. These findings unravel complementary modulation of HUVEC functional phenotypes and signaling molecules involved in angiogenesis by hormone microenvironment and sex-specific factors, and highlight the need for sex-oriented pharmacological targeting of endothelial function.
女性激素和性别特异性因素是已确定的内皮功能决定因素，但其对人类内皮细胞表型的相对贡献尚未明确。我们利用按供体性别进行基因分型的人脐静脉内皮细胞（HUVECs），研究了性别和雌激素类药物对血管生成过程主要步骤以及调控HUVEC代谢和迁移特性的关键蛋白的影响。与男性供体来源的HUVECs（mHUVECs）相比，女性供体来源的HUVECs（fHUVECs）显示出更高的活力（<0.01）和生长速率（<0.01）。尽管fHUVECs中G蛋白偶联雌激素受体（GPER）水平较高（<0.001），但用17β-雌二醇（E2）和选择性GPER激动剂G1（均为1 - 100 nM）处理并未影响HUVEC的活力。在生理条件下，fHUVECs的迁移和形成管状结构的能力高于mHUVECs（<0.05）。尽管基线水平相当，但E2处理（1 - 100 nM）上调糖酵解激活剂PFKFB3的能力在fHUVECs中高于mHUVECs。此外，尽管Src激活水平相当，但粘着斑激酶（FAK）的Y576/577磷酸化在fHUVECs中显著增强（<0.001）。虽然PI3K抑制剂LY294002（25 μM）抑制HUVEC迁移（<0.05），但fHUVECs和mHUVECs中Akt磷酸化水平相当。最后，抑制Y576/577 FAK磷酸化的洋地黄毒苷处理消除了HUVEC迁移中的性别差异。这些发现揭示了激素微环境和性别特异性因素对参与血管生成的HUVEC功能表型和信号分子的互补调节，并强调了针对内皮功能进行性别导向药理学靶向治疗的必要性。
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