Arterioscler Thromb Vasc Biol Pravastatin Enhances Beneficial Effects of Olmesartan on Vascular Injury of Salt-Sensitive Hypertensive Rats, via Pleiotropic Effects by Eiichiro Yamamoto, Takuro Yamashita, Tomoko Tanaka, Keiichiro Kataoka, Yoshiko Tokutomi, Zhong-Fang Lai, Yi-Fei Dong, Shinji Matsuba, Hisao Ogawa, and Shokei Kim-Mitsuyama Arterioscler Thromb Vasc Biol Volume 27(3):556-563 March 1, 2007 Copyright © American Heart Association, Inc. All rights reserved.

Figure 1. Time course of vascular NADPH oxidase (A), superoxide (B), eNOS activity (C), and plasma NO2/NO3 (D) in DS rats. Figure 1. Time course of vascular NADPH oxidase (A), superoxide (B), eNOS activity (C), and plasma NO2/NO3 (D) in DS rats. High Na, 8% NaCl diet; Low Na, 0.3% NaCl diet. Values are mean±SEM (n=5). Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.

Figure 2. Effect of vehicle (V), olmesartan (O), pravastatin (P), combined olmesartan and pravastatin (O+P), and hydralazine (H) on acetylcholine-induced vascular relaxation (A), coronary arterial thickening (B), and perivascular fibrosis (C) of DS rats fed high-salt diet. Figure 2. Effect of vehicle (V), olmesartan (O), pravastatin (P), combined olmesartan and pravastatin (O+P), and hydralazine (H) on acetylcholine-induced vascular relaxation (A), coronary arterial thickening (B), and perivascular fibrosis (C) of DS rats fed high-salt diet. A, Each plot represents mean±SEM (n=8 to 9 per group). Right top panels show representative images of light micrographs of LV coronary arterial thickening and perivascular fibrosis. Original magnification, ×200. Each bar represents mean±SEM (n=4 to 8 per group). L indicates DS rats fed low-salt diet; NS, not significant. Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.

Figure 3. Effect of each treatment on vascular NADPH oxidase (A), p22phox (B), superoxide (C), and eNOS activity (D) of DS rats fed high-salt diet. Figure 3. Effect of each treatment on vascular NADPH oxidase (A), p22phox (B), superoxide (C), and eNOS activity (D) of DS rats fed high-salt diet. Top panels in B and C show representative Western blot and fluorescence photomicrographs, respectively, in each group. #P<0.05, *P<0.01 vs V. Each bar represents mean±SEM. A: L, n=9; V, n=9; H, n=4; O, n=9; P, n=9; O+P, n=8. B: L, n=9; V, n=9; H, n=5; O, n=9; P, n=9; O+P, n=8. C: n=5 per group. D: n=4 to 5 per group. Abbreviations are the same as Figure 2. L indicates DS rats fed low-salt diet. Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.

Figure 4. Effect of each treatment on phospho-akt (p-akt), total akt, phospho-eNOS (p-eNOS), and total eNOS of DS rats fed high-salt diet. Figure 4. Effect of each treatment on phospho-akt (p-akt), total akt, phospho-eNOS (p-eNOS), and total eNOS of DS rats fed high-salt diet. Top panels show representative Western blot in each group. Each bar represents mean±SEM (n=5 per group). Abbreviations are the same as Figure 2. L indicates DS rats fed low-salt diet. Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.

Figure 5. Effect of each treatment on eNOS dimer disruption (A) and DHFR protein levels (B) of DS rats fed high-salt diet. Figure 5. Effect of each treatment on eNOS dimer disruption (A) and DHFR protein levels (B) of DS rats fed high-salt diet. Top panels in A and B show representative Western blot in each group. Each bar represents mean±SEM (n=5 per group). Abbreviations are the same as Figure 2. L indicates DS rats fed low-salt diet. Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.

Figure 6. Effect of each treatment on vascular nitrotyrosine of DS rats fed high-salt diet. Figure 6. Effect of each treatment on vascular nitrotyrosine of DS rats fed high-salt diet. Vascular nitrotyrosine was determined by immunohistochemistry (A) and Western blot analysis (B). Each bar represents mean±SEM (n=5 per group). Abbreviations are the same as Figure 2. L indicates DS rats fed low-salt diet. Eiichiro Yamamoto et al. Arterioscler Thromb Vasc Biol. 2007;27:556-563 Copyright © American Heart Association, Inc. All rights reserved.