title: Early hypercholesterolemia contributes to vasomotor dysfunction and injury associated atherogenesis that can be inhibited by nitric oxide.
abstract: OBJECTIVE
  Atherosclerosis results in vasomotor dysfunction, in part, through impairment of nitric oxide (NO) dependent vasodilation. It is unclear whether blood vessels are dysfunctional in an early environment of hypercholesterolemia alone and if this contributes to the vascular injury response. We hypothesize that early hypercholesterolemia, prior to gross vascular changes, contributes to vasomotor dysfunction and the vascular injury response. The efficacy of NO therapy to protect against the injury response in this setting was also assessed.
  
  
  METHODS
  The effect of oxidized low density lipoprotein (oxLDL) and inducible NO synthase (iNOS) gene transfer on rat aortic smooth muscle cell (SMC) proliferation was measured with (3)H-thymidine incorporation. Common carotid arteries (CCA) from wild-type C57BL6 (WT or C57) and apolipoprotein E deficient (ApoE KO) mice fed normal or Western diets for 6 to 8 weeks were tested for vasomotor function using an arteriograph system. Studies were repeated after CCA injury. The effect of iNOS gene transfer on morphometry by histology and vasomotor responses in injured CCAs in ApoE KO was examined.
  
  
  RESULTS
  OxLDL increased SMC proliferation by >50%. In SMC expressing iNOS, NO production was unaffected by oxLDL and reduced oxLDL and still inhibited SMC proliferation. Endothelium dependent vasorelaxation was reduced in uninjured CCAs from ApoE KO and C57 mice on the Western vs normal diet (ApoE 39% ± 2% vs 55% ± 13%; C57 50% ± 13% vs 76% ± 5%, P < .001) and was increased with longer durations of hypercholesterolemia. Endothelium-dependent and independent vasodilator responses were severely disrupted in C57 and ApoE KO mice 2 weeks following CCA injury but both recovered by 4 weeks. CCA injury in ApoE KO mice resulted in the formation of atheromatous lesions while C57 mice showed no change (intima 27,795 ± 1829 vs 237 ± 28 μm(2); media 46,306 ± 2448 vs 11,714 ± 392 μm(2), respectively; P < .001). This structural change in the ApoE KO reduced distensibility and increased stiffness. Finally, iNOS gene transfer to injured CCA in ApoE KO mice dramatically reduced atheromatous neointimal lesion formation.
  
  
  CONCLUSIONS
  Early hypercholesterolemia impairs endothelial function, with severity being related to duration and magnitude of hypercholesterolemia. Severe hypercholesterolemia leads to atheromatous lesion formation following injury and stresses the role of vascular injury in atherogenesis and suggests different mechanisms are involved in endothelial dysfunction and the injury response. Despite these changes, iNOS gene transfer still effectively inhibits atheroma formation. These findings support early correction of hypercholesterolemia and emphasize the potential role for NO based therapies in disease states.
authors.affiliation: Division of Vascular Surgery, Department of Surgery, University of Pittsburgh, Pittsburgh, Pa 15213, USA.
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Raman
authors.forename: Robin E
authors.initials: RE
authors.lastname: Gandley
authors.forename: Jennifer
authors.initials: J
authors.lastname: Rohland
authors.forename: Mazen S
authors.initials: MS
authors.lastname: Zenati
authors.forename: Edith
authors.initials: E
authors.lastname: Tzeng
mesh_terms: D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D050197:Atherosclerosis / Q000209:etiology / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D020212:Carotid Artery Injuries / Q000150:complications* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D017536:Carotid Artery, Common / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D049109:Cell Proliferation; D002478:Cells, Cultured; D002784:Cholesterol / Q000097:blood; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D006937:Hypercholesterolemia / Q000150:complications* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D008077:Lipoproteins, LDL / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000503:physiopathology; D032389:Myocytes, Smooth Muscle / Q000201:enzymology; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D013997:Time Factors; D014162:Transfection; D014280:Triglycerides / Q000097:blood; D014664:Vasodilation / Q000187:drug effects; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D001057:Apolipoproteins E; D008077:Lipoproteins, LDL; D014280:Triglycerides; D014665:Vasodilator Agents; C099252:oxidized low density lipoprotein; D009569:Nitric Oxide; D002784:Cholesterol; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II