Ischemic stroke disrupts the endothelial glycocalyx through activation of proHPSE via acrolein exposure

J Biol Chem. 2020 Dec 25;295(52):18614-18624. doi: 10.1074/jbc.RA120.015105. Epub 2020 Oct 30.


Infiltration of peripheral immune cells after blood-brain barrier dysfunction causes severe inflammation after a stroke. Although the endothelial glycocalyx, a network of membrane-bound glycoproteins and proteoglycans that covers the lumen of endothelial cells, functions as a barrier to circulating cells, the relationship between stroke severity and glycocalyx dysfunction remains unclear. In this study, glycosaminoglycans, a component of the endothelial glycocalyx, were studied in the context of ischemic stroke using a photochemically induced thrombosis mouse model. Decreased levels of heparan sulfate and chondroitin sulfate and increased activity of hyaluronidase 1 and heparanase (HPSE) were observed in ischemic brain tissues. HPSE expression in cerebral vessels increased after stroke onset and infarct volume greatly decreased after co-administration of N-acetylcysteine + glycosaminoglycan oligosaccharides as compared with N-acetylcysteine administration alone. These results suggest that the endothelial glycocalyx was injured after the onset of stroke. Interestingly, scission activity of proHPSE produced by immortalized endothelial cells and HEK293 cells transfected with hHPSE1 cDNA were activated by acrolein (ACR) exposure. We identified the ACR-modified amino acid residues of proHPSE using nano LC-MS/MS, suggesting that ACR modification of Lys139 (6-kDa linker), Lys107, and Lys161, located in the immediate vicinity of the 6-kDa linker, at least in part is attributed to the activation of proHPSE. Because proHPSE, but not HPSE, localizes outside cells by binding with heparan sulfate proteoglycans, ACR-modified proHPSE represents a promising target to protect the endothelial glycocalyx.

Keywords: acrolein; chondroitin sulfate; glycocalyx; heparan sulfate; heparanase; hyaluronan; hyaluronidase; ischemic stroke; stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrolein / pharmacology*
  • Amino Acid Sequence
  • Animals
  • Brain Ischemia / etiology
  • Brain Ischemia / metabolism
  • Brain Ischemia / pathology*
  • Chondroitin Sulfates / metabolism
  • Disease Models, Animal
  • Endothelium, Vascular / metabolism
  • Endothelium, Vascular / pathology*
  • Glucuronidase / chemistry
  • Glucuronidase / genetics
  • Glucuronidase / metabolism*
  • Glycocalyx / metabolism
  • Glycocalyx / pathology*
  • Heparitin Sulfate / metabolism
  • Humans
  • Hyaluronoglucosaminidase / metabolism
  • Ischemic Stroke / etiology
  • Ischemic Stroke / metabolism
  • Ischemic Stroke / pathology*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Photochemistry
  • Protein Conformation


  • Acrolein
  • Chondroitin Sulfates
  • Heparitin Sulfate
  • heparanase
  • Glucuronidase
  • Hyaluronoglucosaminidase

Associated data

  • PDB/5LA4