Endothelial and non-endothelial actions of estrogen receptor subtypes in male and female mouse Aorta

IF 3.7 3区医学 Q2 PHARMACOLOGY & PHARMACY

Vascular pharmacology Pub Date : 2025-12-01 Epub Date: 2025-11-08 DOI:10.1016/j.vph.2025.107559

Basant Elsaid , Irakli Kopaliani , Birgit Zatschler , Ansam Seif , Stephan Speier , Andreas Deussen

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Abstract

Estrogen's role in cardiovascular health remains inconsistent. This study investigates the non-genomic vasorelaxant effects of 17β-estradiol, focusing on sex-specific differences in estrogen receptor (ER) subtypes (ERα, ERβ and GPER), endothelial function, and phosphoinositide 3-kinase (PI3K) pathway.

Ex vivo experiments using thoracic aorta from male and female wild-type mice assessed ER expression by immunofluorescence and vascular reactivity by wire myography. Vasorelaxant responses to 17β-estradiol and ER-selective agonists (PPT, DPN and G1) were examined. Some experiments were conducted in the presence of pharmacological inhibitors targeting endothelium-dependent relaxation pathways (L-NMMA, indomethacin and apamin/charybdotoxin), as well as PI3K inhibitor LY294002. In vitro, primary mouse aortic smooth muscle cells (MASMCs) were used to assess acute effects of 17β-estradiol on myosin light chain (MLC) and Akt phosphorylation via western blot.

ER subtypes were observed in the endothelial and medial regions of the vessel wall. 17β-estradiol induced similar vasorelaxation in both sexes, with subtle differences in endothelial ERβ and GPER. Nitric oxide synthase (NOS) inhibition attenuated vasorelaxation, with modest sex-specific differences in ERβ. PI3K inhibition augmented vasorelaxation regardless of endothelial integrity, exhibiting slight sex variability across ER subtypes. MASMCs showed a near-significant increase in MLC phosphorylation with 17β-estradiol, without significant Akt activation.

In conclusion, estrogen induces NOS dependent vasorelaxation in mouse aorta, with no sex differences. However, subtle sex differences among ER subtypes were observed in the contribution of endothelium, NOS and PI3K pathways. The PI3K pathway may attenuate vasorelaxation by promoting MLC phosphorylation in vascular smooth muscle, and its inhibition unmasks the vasorelaxant potential of 17β-estradiol.

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雌激素受体亚型在雄性和雌性小鼠主动脉中的内皮和非内皮作用。

雌激素在心血管健康中的作用仍然不一致。本研究探讨了17β-雌二醇的非基因组性血管松弛作用，重点研究了雌激素受体（ER）亚型（ERα、ERβ、GPER）、内皮功能和磷酸肌苷激酶（PI3K）信号通路的性别差异。利用雄性和雌性野生型小鼠胸主动脉进行离体实验，免疫荧光法检测ER表达，钢丝肌图检测血管反应性。观察17β-雌二醇和er选择性激动剂（PPT、DPN、G1）的血管松弛反应。一些实验是在针对内皮依赖性松弛途径的药物抑制剂- l - nmma、吲哚美辛和apamin/charybdotoxin-以及PI3K途径抑制剂LY294002 hydrochloride存在的情况下进行的。在体外，采用小鼠主动脉平滑肌细胞（MASMCs），通过western blot评估17β-雌二醇对肌球蛋白轻链（MLC）和Akt磷酸化的短期剂量依赖性影响。内皮和血管平滑肌层均检测到ER亚型。17β-雌二醇诱导血管松弛，性别差异不显著。一氧化氮合酶（NOS）的抑制降低了血管舒张，内质网亚型之间存在适度的性别特异性差异。PI3K抑制增强了这种血管松弛，无论内皮完整性如何，在不同的ER亚型中表现出轻微的性别特异性差异。17β-雌二醇对MASMCs的MLC磷酸化有增加的趋势，但没有明显的Akt激活。综上所述，雌激素诱导小鼠主动脉NOS依赖性血管松弛，性别差异不显著。然而，内质网亚型对NOS和PI3K通路的贡献存在细微的性别差异。PI3K通路可能通过促进血管平滑肌中MLC的磷酸化而减弱血管松弛作用，其抑制作用揭示了雌激素的血管松弛潜能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Vascular pharmacology 医学-药学

CiteScore

6.60

自引率

2.50%

发文量

153

审稿时长

31 days

期刊介绍： Vascular Pharmacology publishes papers, which contains results of all aspects of biology and pharmacology of the vascular system. Papers are encouraged in basic, translational and clinical aspects of Vascular Biology and Pharmacology, utilizing approaches ranging from molecular biology to integrative physiology. All papers are in English. The Journal publishes review articles which include vascular aspects of thrombosis, inflammation, cell signalling, atherosclerosis, and lipid metabolism.