title: Reactive oxygen species mediate high glucose-induced heparanase-1 production and heparan sulphate proteoglycan degradation in human and rat endothelial cells: a potential role in the pathogenesis of atherosclerosis.
abstract: OBJECTIVE
  The content of heparan sulphate is reduced in the endothelium under hyperglycaemic conditions and may contribute to the pathogenesis of atherosclerosis. Heparanase-1 (HPR1) specifically degrades heparan sulphate proteoglycans. We therefore sought to determine whether: (1) heparan sulphate reduction in endothelial cells is due to increased HPR1 production through increased reactive oxygen species (ROS) production; and (2) HPR1 production is increased in vivo in endothelial cells under hyperglycaemic and/or atherosclerotic conditions.
  
  
  METHODS
  HPR1 mRNA and protein levels in endothelial cells were analysed by RT-PCR and Western blot or HPR1 enzymatic activity assay, respectively. Cell surface heparan sulphate levels were analysed by FACS. HPR1 in the artery from control rats and a rat model of diabetes, and from patients under hyperglycaemic and/or atherosclerotic conditions was immunohistochemically examined.
  
  
  RESULTS
  High-glucose-induced HPR1 production and heparan sulphate degradation in three human endothelial cell lines, both of which were blocked by ROS scavengers, glutathione and N-acetylcysteine. Exogenous H(2)O(2) induced HPR1 production, subsequently leading to decreased cell surface heparan sulphate levels. HPR1 content was significantly increased in endothelial cells in the arterial walls of a rat model of diabetes. Clinical studies revealed that HPR1 production was increased in endothelial cells under hyperglycaemic conditions, and in endothelial cells and macrophages in atherosclerotic lesions.
  
  
  CONCLUSIONS
  Hyperglycaemia induces HPR1 production and heparan sulphate degradation in endothelial cells through ROS. HPR1 production is increased in endothelial cells from a rat model of diabetes, and in macrophages in the atherosclerotic lesions of diabetic and non-diabetic patients. Increased HPR1 production may contribute to the pathogenesis and progression of atherosclerosis.
authors.affiliation: Department of General Surgery, Rush University Medical Center, 1653 W Congress Parkway, Chicago, IL 60612, USA.
authors.forename: G
authors.initials: G
authors.lastname: Rao
authors.forename: H G
authors.initials: HG
authors.lastname: Ding
authors.forename: W
authors.initials: W
authors.lastname: Huang
authors.forename: D
authors.initials: D
authors.lastname: Le
authors.forename: J B
authors.initials: JB
authors.lastname: Maxhimer
authors.forename: A
authors.initials: A
authors.lastname: Oosterhof
authors.forename: T
authors.initials: T
authors.lastname: van Kuppevelt
authors.forename: H
authors.initials: H
authors.lastname: Lum
authors.forename: E J
authors.initials: EJ
authors.lastname: Lewis
authors.forename: V
authors.initials: V
authors.lastname: Reddy
authors.forename: R A
authors.initials: RA
authors.lastname: Prinz
authors.forename: X
authors.initials: X
authors.lastname: Xu
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D050197:Atherosclerosis / Q000209:etiology* / Q000378:metabolism*; D002460:Cell Line; D003920:Diabetes Mellitus / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005947:Glucose / Q000494:pharmacology; D005966:Glucuronidase / Q000378:metabolism*; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006943:Hyperglycemia / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013311:Streptozocin / Q000009:adverse effects; D000099175:Heparanase
chemical_list: D005947:Glucose; D005966:Glucuronidase; D019812:Heparan Sulfate Proteoglycans; D006861:Hydrogen Peroxide; D017382:Reactive Oxygen Species; D013311:Streptozocin; D000099175:Heparanase