Journal: Circulation

Article Title: Aldosterone Enhances Ischemia-Induced Neovascularization Through Angiotensin II–Dependent Pathway

doi: 10.1161/01.cir.0000127112.36796.9b

Figure Lengend Snippet: Figure 1. Representative photomicro- graphs and quantitative evaluation of microangiography (A), capillary density (B; capillary appears in white, and arrows indicate representative examples of fibronectin-positive capillaries), and foot perfusion (C). D, Representative pho- tomicrographs of Western blot and quantitative evaluation of VEGF protein content. Values are meanSEM; n7. *P0.05 vs control mice; #P0.05 vs aldosterone-treated mice; †P0.05 vs nonischemic control. C indicates untreated animals; A, aldosterone-treated mice; AS, aldosterone and spironolac- tone–treated mice; AV, aldosterone and valsartan–treated mice; Aa-VEGF, aldo- sterone and neutralizing VEGF–treated mice; Mb, membrane stained with pon- ceau red; N. Isch., nonischemic; Isch., ischemic; Cont, control; and Aldo, aldosterone.

Article Snippet: C57Bl/6 mice (aged 10 weeks; Iffa Creddo, Lyon, France) were anesthetized by isoflurane inhalation, and unilateral hindlimb ischemia was induced by ligature (tied for complete occlusion) on the right femoral artery, as previously described).4,6 Mice were then randomly assigned to one of the following groups (n 7): (1) control group: vehicle (1% ethanol in drinking water); (2) mice receiving the mineralocorticoid receptor blocker spironolactone (spironolactone dissolved in ethanol at 25 mg/mL and added to drinking water; mice then received 20 mg/kg per day; Sigma); (3) mice treated with the AT1 receptor blocker valsartan (drinking water, 20 mg/kg per day; Sigma); (4) aldosterone (4.5 g/d by osmotic minipumps implanted subcutaneously in the back of the mice; model 2004, Alza Corp); (5) aldosterone plus spironolactone; (6) aldosterone plus valsartan; and (7) aldosterone plus a-VEGF neutralizing antibody (10 g IP twice per week; R&D Systems).

Techniques: Western Blot, Control, Membrane, Staining