Azelnidipine suppresses the progression of aortic aneurysm in wild mice model through anti-inflammatory effects Hirotsugu Kurobe, MD, PhD, Yuki Matsuoka,

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Azelnidipine suppresses the progression of aortic aneurysm in wild mice model through anti-inflammatory effects Hirotsugu Kurobe, MD, PhD, Yuki Matsuoka, BS, Yoichiro Hirata, MD, PhD, Noriko Sugasawa, BS, Mark W. Maxfield, MD, Masataka Sata, MD, PhD, Tetsuya Kitagawa, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 146, Issue 6, Pages 1501-1508 (December 2013) DOI: 10.1016/j.jtcvs.2013.02.073 Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Blood pressure measurements. A, Changes over time in systolic blood pressure during the infusion of angiotensin II and BAPN. There is no significant difference between groups (n = 12 in control group, n = 11 in azelnidipine group). B, Maximum blood pressure of each mouse after 4 weeks of combination infusion therapy (n = 12 in control group; n = 11 in azelnidipine group). BAPN, β-Aminopropionitrile. The Journal of Thoracic and Cardiovascular Surgery 2013 146, 1501-1508DOI: (10.1016/j.jtcvs.2013.02.073) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Morphologic changes of aorta after combination infusion with angiotensin II and BAPN. A, Representative images of aortic aneurysm. Thoracic aortic aneurysm in upper panel. Abdominal aortic aneurysm in lower panel. BAPN, β-Aminopropionitrile; TAA, thoracic aortic aneurysm; AAA, abdominal aortic aneurysm; BT, brachiocephalic trunk; CA, carotid artery; SA, subclavian artery; RA, renal artery; Ao, aorta; Lt, left; Rt, right. B to D, Aortic ratio in each mouse. B, All aortic lesions. C, Thoracic aortic lesions. D, Abdominal aortic lesions. (n = 12 in control group, n = 11 in azelnidipine group.) The Journal of Thoracic and Cardiovascular Surgery 2013 146, 1501-1508DOI: (10.1016/j.jtcvs.2013.02.073) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Histologic analysis of aorta. A, Hematoxylin and eosin (H&E) staining of aorta. LPF, Low-power field (bar shows length in 500 μm); HPF, high-power field (bar shows 100 μm of length). B, Elastica van Gieson (EVG) staining of aorta (bar shows 100 μm). The Journal of Thoracic and Cardiovascular Surgery 2013 146, 1501-1508DOI: (10.1016/j.jtcvs.2013.02.073) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Mac-3 immunohistochemical staining of aorta. A, Representative images of Mac-3 immunohistochemical staining (bar shows 50 μm of length). B, The number of Mac-3-positive cells in aortic lesions was counted in the control group (treated with a combination infusion of angiotensin II and BAPN), azelnidipine group (treated with the combination infusion plus azelnidipine administration), and normal group (nontreated). Left panel: aortic wall; right panel: periaortic adipose tissue; n = 7 in each group. BAPN, β-Aminopropionitrile. The Journal of Thoracic and Cardiovascular Surgery 2013 146, 1501-1508DOI: (10.1016/j.jtcvs.2013.02.073) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Gene expression levels of TNF-α, MMP-2, MMP-9, and SIRT-1 in aortic wall. Real-time PCR analyses of (A) TNF-α, (B) MMP-2, (C) MMP-9, and (D) SIRT-1 were performed in mice treated with combination infusion of angiotensin II and BAPN only (control group), mice administered azelnidipine and combination infusion (azelnidipine group), and mice treated with placebo vehicle (normal group). Values are expressed as mean ± standard error (n = 7 in each group). TNF-α, Tumor necrosis factor-alpha; MMP, matrix metalloproteinase; RT-PCR, reverse-transcriptase polymerase chain reaction; BAPN, β-aminopropionitrile. The Journal of Thoracic and Cardiovascular Surgery 2013 146, 1501-1508DOI: (10.1016/j.jtcvs.2013.02.073) Copyright © 2013 The American Association for Thoracic Surgery Terms and Conditions