Neutrophil extracellular traps boost laser-induced mouse choroidal neovascularization through the activation of the choroidal endothelial cell TLR4/HIF-1α pathway

FEBS J. 2023 Nov;290(22):5395-5410. doi: 10.1111/febs.16928. Epub 2023 Sep 4.

Abstract

Choroidal neovascularization (CNV) is characterized by the infiltration of immune cells, particularly neutrophils. Neutrophil extracellular trap (NET) facilitates the angiogenesis of pulmonary endothelial cells via activating Toll-like receptor 4 (TLR4). TLR4 promotes the expression of transcription factor hypoxia inducible factor-1α (HIF-1α), which promotes inflammation and angiogenesis via the up-regulation of metalloproteinase-9 (MMP-9) and interleukin-1β (IL-1β). In the present study, we aimed to identify the formation of NET and its role in CNV. Our results showed that NET levels were increased in a mouse laser-induced CNV model via oxidative stress, whereas the inhibition of NET alleviated CNV. In vitro, NET activated the TLR4/HIF-1α pathway in human choroidal endothelial cells (HCECs). Additionally, NET increased the transcription and expression of MMP-9 and IL-1β in HCECs via activating the TLR4/HIF-1α pathway. Meanwhile, NET promoted the inflammatory response accompanied by the proliferation, migration and tube formation of HCECs in a MMP-9- and IL-1β-dependent manner. In conclusion, NET was up-regulated in CNV and promoted the formation of CNV via activating the TLR4/HIF-1α pathway in choroidal endothelial cells. Our data uncovered the novel role of NET in promoting the formation of CNV. The underlying mechanism of NET could be targeted to delay the process of CNV.

Keywords: Toll-like receptor 4; choroidal endothelial cells; choroidal neovascularization; hypoxia inducible factor-1α; neutrophil extracellular trap.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Choroidal Neovascularization* / genetics
  • Disease Models, Animal
  • Endothelial Cells / metabolism
  • Extracellular Traps*
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Lasers
  • Matrix Metalloproteinase 9 / genetics
  • Mice
  • Toll-Like Receptor 4 / genetics
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Toll-Like Receptor 4
  • Matrix Metalloproteinase 9
  • Vascular Endothelial Growth Factor A
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • TLR4 protein, human