title: Activation of TGF-β receptors and Smad proteins by atorvastatin is related to reduced atherogenesis in ApoE/LDLR double knockout mice.
abstract: OBJECTIVE
  Transforming growth factor-beta (TGF-β) plays important role in atherogenesis via TGF-β receptors and Smad proteins, which determine its signaling activity. In this study, we hypothesized, whether non-lipid related effects of atorvastatin, affect both endoglin/ALK-5/Smad2/eNOS and/or endoglin/ALK-1/Smad1/VEGF previously proposed pathways in ApoE/LDLR double knockout mice.
  
  
  METHODS
  ApoE/LDLR double knockout mice were divided into two groups. The chow group (CHOW) (n =8) was fed with chow diet, while in the atorvastatin group (ATV) (n =8) atorvastatin was added to the chow diet at dose 50 mg/kg/day. Biochemical analyses of lipid profile, lesion area measurement, immunohistochemistry and Western blot analysis of endoglin, ALK-1, 5, phosphorylated and non-phosphorylated forms Smad-1, 2, VEGF and eNOS proteins in mice aorta were performed.
  
  
  RESULTS
  Biochemical analysis of blood serum and morphometric analysis of aortic lesion size showed that atorvastatin treatment resulted in a significant increase of cholesterol levels and simultaneously in reduced lesion size in aortic sinus when compared to CHOW mice. Western blot analysis revealed that atorvastatin treatment significantly increase the expressions of endoglin by 102%, ALK-1 by 113%, ALK-5 by 296%, pSmad-1 by 202%, pSmad-2 by 34%, VEGF by 68% and eNOS by 687% as compared with CHOW mice. Immunofluorescence staining revealed endoglin coexpression with all studied markers that were increased by atorvastatin treatment mainly in endothelial cells covering atherosclerotic plaques.
  
  
  CONCLUSIONS
  This study shows that atorvastatin treatment increases the expression of endoglin, ALK-1, ALK-5, phosphorylated forms of Smad1 and Smad2, VEGF and eNOS and reduces atherosclerotic lesion size beyond its lipid lowering effects. Therefore, we propose that endoglin related receptors and signal transducers might play protective role in atherogenesis.
authors: Department of Biological and Medical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, Czech Republic.
authors: Lenka
authors: L
authors: Vecerova
authors: Zbynek
authors: Z
authors: Strasky
authors: Jana
authors: J
authors: Rathouska
authors: Martina
authors: M
authors: Slanarova
authors: Eva
authors: E
authors: Brcakova
authors: Stanislav
authors: S
authors: Micuda
authors: Petr
authors: P
authors: Nachtigal
mesh_terms: D000818:Animals; D000924:Anticholesteremic Agents / Q000627:therapeutic use; D001057:Apolipoproteins E / Q000502:physiology*; D050197:Atherosclerosis / Q000378:metabolism / Q000517:prevention & control*; D000069059:Atorvastatin; D015153:Blotting, Western; D002784:Cholesterol / Q000378:metabolism; D000071063:Endoglin; D005260:Female; D006538:Heptanoic Acids / Q000627:therapeutic use*; D007124:Immunoenzyme Techniques; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011758:Pyrroles / Q000627:therapeutic use*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D011973:Receptors, LDL / Q000502:physiology*; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D051898:Smad1 Protein / Q000378:metabolism*; D051899:Smad2 Protein / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D000924:Anticholesteremic Agents; D001057:Apolipoproteins E; D000071063:Endoglin; C502004:Eng protein, mouse; D006538:Heptanoic Acids; D047908:Intracellular Signaling Peptides and Proteins; D011758:Pyrroles; D011973:Receptors, LDL; D018125:Receptors, Transforming Growth Factor beta; D051898:Smad1 Protein; C497401:Smad1 protein, mouse; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D002784:Cholesterol; D000069059:Atorvastatin; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625842:Tgfbr1 protein, mouse