title: Endothelial nitric oxide synthase and superoxide mediate hemodynamic initiation of intracranial aneurysms.
abstract: BACKGROUND
  Hemodynamic insults at arterial bifurcations are believed to play a critical role in initiating intracranial aneurysms. Recent studies in a rabbit model indicate that aneurysmal damage initiates under specific wall shear stress conditions when smooth muscle cells (SMCs) become pro-inflammatory and produce matrix metalloproteinases (MMPs). The mechanisms leading to SMC activation and MMP production during hemodynamic aneurysm initiation are unknown. The goal is to determine if nitric oxide and/or superoxide induce SMC changes, MMP production and aneurysmal remodeling following hemodynamic insult.
  
  
  METHODS
  Bilateral common carotid artery ligation was performed on rabbits (n = 19, plus 5 sham operations) to induce aneurysmal damage at the basilar terminus. Ligated animals were treated with the nitric oxide synthase (NOS) inhibitor LNAME (n = 7) or the superoxide scavenger TEMPOL (n = 5) and compared to untreated animals (n = 7). Aneurysm development was assessed histologically 5 days after ligation. Changes in NOS isoforms, peroxynitrite, reactive oxygen species (ROS), MMP-2, MMP-9, and smooth muscle α-actin were analyzed by immunohistochemistry.
  
  
  RESULTS
  LNAME attenuated ligation-induced IEL loss, media thinning and bulge formation. In untreated animals, immunofluorescence showed increased endothelial NOS (eNOS) after ligation, but no change in inducible or neuronal NOS. Furthermore, during aneurysm initiation ROS increased in the media, but not the intima, and there was no change in peroxynitrite. In LNAME-treated animals, ROS production did not change. Together, this suggests that eNOS is important for aneurysm initiation but not by producing superoxide. TEMPOL treatment reduced aneurysm development, indicating that the increased medial superoxide is also necessary for aneurysm initiation. LNAME and TEMPOL treatment in ligated animals restored α-actin and decreased MMPs, suggesting that eNOS and superoxide both lead to SMC de-differentiation and MMP production.
  
  
  CONCLUSIONS
  Aneurysm-inducing hemodynamics lead to increased eNOS and superoxide, which both affect SMC phenotype, increasing MMP production and aneurysmal damage.
authors.affiliation: Toshiba Stroke and Vascular Research Center and Department of Mechanical and Aerospace Engineering, State University of New York, Buffalo, New York, United States of America.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Liaw
authors.affiliation: Toshiba Stroke and Vascular Research Center and Department of Neurosurgery, State University of New York, Buffalo, New York, United States of America.
authors.forename: Jennifer M Dolan
authors.initials: JM
authors.lastname: Fox
authors.affiliation: Toshiba Stroke and Vascular Research Center and Departments Neurosurgery and Radiology, State University of New York, Buffalo, New York, United States of America.
authors.forename: Adnan H
authors.initials: AH
authors.lastname: Siddiqui
authors.affiliation: Toshiba Stroke and Vascular Research Center and Departments of Mechanical and Aerospace Engineering, Neurosurgery, and Biomedical Engineering, State University of New York, Buffalo, New York, United States of America.
authors.forename: Hui
authors.initials: H
authors.lastname: Meng
authors.affiliation: Toshiba Stroke and Vascular Research Center and Department Pathology and Anatomical Sciences, State University of New York, Buffalo, New York, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Kolega
mesh_terms: D000818:Animals; D003497:Cyclic N-Oxides / Q000008:administration & dosage*; D004195:Disease Models, Animal; D005260:Female; D002532:Intracranial Aneurysm / Q000378:metabolism / Q000473:pathology*; D053504:Matrix Metalloproteinases, Membrane-Associated / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000473:pathology; D019331:NG-Nitroarginine Methyl Ester / Q000008:administration & dosage*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D011500:Protein Synthesis Inhibitors / Q000008:administration & dosage*; D011817:Rabbits; D017382:Reactive Oxygen Species / Q000378:metabolism; D013113:Spin Labels; D013481:Superoxides / Q000378:metabolism*
chemical_list: D003497:Cyclic N-Oxides; D011500:Protein Synthesis Inhibitors; D017382:Reactive Oxygen Species; D013113:Spin Labels; D013481:Superoxides; D019001:Nitric Oxide Synthase; D053504:Matrix Metalloproteinases, Membrane-Associated; C001803:tempol; D019331:NG-Nitroarginine Methyl Ester