Aspirin restores endothelial function by mitigating 17β-estradiol-induced α-SMA accumulation and autophagy inhibition via Vps15 scaffold regulation of Beclin-1 phosphorylation

Life Sci. 2020 Oct 15:259:118383. doi: 10.1016/j.lfs.2020.118383. Epub 2020 Sep 4.

Abstract

Aims: Previous studies have shown that the widespread use of estrogen preparations can cause adverse outcomes such as thrombosis and cardiovascular disease. Autophagy is a biochemical process necessary to maintain cell homeostasis. The present study investigated whether E-2 mediates autophagy-induced endothelial cell dysfunction. The role of aspirin in this process was then studied.

Main methods: Western blot, fluorescence microscopy, electron transmission microscopy, plasma construction and transfection, vasoreactivity study in wire myograph are all used in this study.

Key findings: We found that E-2 activated the PI3K/mTOR signaling pathway and inhibited the formation of the Atg14L-Beclin1-Vps34-Vps15 complex, thereby inhibiting autophagy. Aspirin promoted Beclin1 phosphorylation in autophagy initiation complexes and enhanced autophagy. Furthermore, E-2 treatment of HAECs resulted in endothelial dysfunction by inhibiting autophagy and leading to accumulation of α-smooth muscle actin (α-SMA). E-2 inhibited the activation of eNOS and reduced the expression of eNOS protein. In the mouse aortic vascular function test, E-2 disrupted endothelium-dependent vasodilation. An α-SMA-shRNA lentivirus eliminated the disruption to endothelium-dependent vasodilation by E-2. Aspirin inhibited α-SMA accumulation by enhancing autophagy, reversed endothelial functional impairment caused by E-2, and promoted endothelium-dependent vasodilation.

Significance: This study provides new evidence that E-2 inhibits autophagy and induces abnormal accumulation of α-SMA, resulting in endothelial cell dysfunction and affecting vasodilation. Aspirin can effectively restore the endothelial cell function disrupted E-2.

Keywords: Aspirin; Autophagy; Endothelial dysfunction; Estrogen-2; Vasodilatation; α-SMA.

MeSH terms

  • Actins / metabolism*
  • Animals
  • Aspirin / pharmacology*
  • Autophagy / drug effects*
  • Blotting, Western
  • Cells, Cultured
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / ultrastructure
  • Estradiol / metabolism*
  • Female
  • Humans
  • Immunoprecipitation
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Electron, Transmission
  • Phosphorylation / drug effects
  • Vacuolar Sorting Protein VPS15 / metabolism*

Substances

  • Actins
  • alpha-smooth muscle actin, mouse
  • Estradiol
  • Vacuolar Sorting Protein VPS15
  • Aspirin