title: Imaging of Intracellular and Extracellular ROS Levels in Atherosclerotic Mouse Aortas Ex Vivo: Effects of Lipid Lowering by Diet or Atorvastatin.
abstract: OBJECTIVE
  The first objective was to investigate if intracellular and extracellular levels of reactive oxygen species (ROS) within the mouse aorta increase before or after diet-induced lesion formation. The second objective was to investigate if intracellular and extracellular ROS correlates to cell composition in atherosclerotic lesions. The third objective was to investigate if intracellular and extracellular ROS levels within established atherosclerotic lesions can be reduced by lipid lowering by diet or atorvastatin.
  
  
  RESULTS
  To address our objectives, we established a new imaging technique to visualize and quantify intracellular and extracellular ROS levels within intact mouse aortas ex vivo. Using this technique, we found that intracellular, but not extracellular, ROS levels increased prior to lesion formation in mouse aortas. Both intracellular and extracellular ROS levels were increased in advanced lesions. Intracellular ROS correlated with lesion content of macrophages. Extracellular ROS correlated with lesion content of smooth muscle cells. The high levels of ROS in advanced lesions were reduced by 5 days high dose atorvastatin treatment but not by lipid lowering by diet. Atorvastatin treatment did not affect lesion inflammation (aortic arch mRNA levels of CXCL 1, ICAM-1, MCP-1, TNF-α, VCAM, IL-6, and IL-1β) or cellular composition (smooth muscle cell, macrophage, and T-cell content).
  
  
  CONCLUSIONS
  Aortic levels of intracellular ROS increase prior to lesion formation and may be important in initiation of atherosclerosis. Our results suggest that within lesions, macrophages produce mainly intracellular ROS whereas smooth muscle cells produce extracellular ROS. Short term atorvastatin treatment, but not lipid lowering by diet, decreases ROS levels within established advanced lesions; this may help explain the lesion stabilizing and anti-inflammatory effects of long term statin treatment.
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Matias
authors.initials: M
authors.lastname: Ekstrand
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Gustafsson Trajkovska
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Jeanna
authors.initials: J
authors.lastname: Perman-Sundelin
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Per
authors.initials: P
authors.lastname: Fogelstrand
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Adiels
authors.affiliation: Department of Pathology, Malmö University Hospital, Lund University, SE-205 02, Malmoe, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Johansson
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Lillemor
authors.initials: L
authors.lastname: Mattsson-Hultén
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Jan
authors.initials: J
authors.lastname: Borén
authors.affiliation: Department of Molecular and Clinical Medicine/Wallenberg Laboratory, University of Gothenburg and Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden; Department of Oncology, Sahlgrenska University Hospital, SE-413 45, Gothenburg, Sweden.
authors.forename: Max
authors.initials: M
authors.lastname: Levin
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001011:Aorta / Q000378:metabolism* / Q000473:pathology; D001057:Apolipoproteins E / Q000235:genetics; D000069059:Atorvastatin / Q000494:pharmacology*; D001562:Benzimidazoles; D003324:Coronary Artery Disease / Q000473:pathology*; D018752:Diet, Fat-Restricted; D005260:Female; D008055:Lipids / Q000097:blood; D008163:Luminescent Measurements; D051379:Mice; D018345:Mice, Knockout; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001057:Apolipoproteins E; D001562:Benzimidazoles; D008055:Lipids; D017382:Reactive Oxygen Species; D000069059:Atorvastatin; C017807:bisbenzimide ethoxide trihydrochloride