Aldosterone enhances ischemia-induced neovascularization through angiotensin II-dependent pathway

Background-We analyzed the role of aldosterone in ischemia-induced neovascularization and the involvement of angiotensin II (Ang II) signaling in this effect. Methods and Results-Ischemia was induced by right femoral artery ligature in mice treated or not with aldosterone (4.5 mug/day), aldosterone plus spironolactone (aldosterone receptor blocker; 20 mg/kg per day), or aldosterone plus valsartan (angiotensin type 1 [AT(1)] receptor blocker; 20 mg/kg per day). After 21 days, neovascularization was evaluated by microangiography, capillary density measurement, and laser-Doppler perfusion imaging. Protein level of vascular endothelial growth factor (VEGF) was determined by Western blot analysis in hindlimbs. mRNA levels of renin-angiotensin system components were also assessed by semiquantitative reverse transcription-polymerase chain reaction. Angiographic score, capillary number, and foot perfusion were improved in ischemic/nonischemic leg ratio by 1.4-, 1.5-, and 1.4-fold, respectively, in aldosterone-treated mice compared with controls (P < 0.05). Aldosterone proangiogenic effect was associated with 2.3-fold increase in VEGF protein content (P < 0.05). Treatments with spironolactone or with neutralizing VEGF antibody hampered the proangiogenic effect of aldosterone (P < 0.05 versus aldosterone-treated mice). Interestingly, AT(1) receptor blockade completely abrogated the aldosterone proangiogenic effect, emphasizing the involvement of Ang II-related pathway in aldosterone-induced vessel growth. In this view, angiotensinogen mRNA content was 2.2-fold increased in aldosterone-treated mice in reference to controls (P < 0.05), whereas that of renin, angiotensin-converting enzyme, and AT1 receptor subtype was unaffected. Aldosterone treatment also decreased AT(2) mRNA content by 2-fold (P < 0.05 versus controls), suggesting that aldosterone may switch the Ang II pathway toward activation of vessel growth. Conclusions-This study shows for the first time that aldosterone increases neovascularization in the setting of ischemia through activation of Ang II signaling.