Aging exacerbates neointimal formation, and increases proliferation and reduces susceptibility to apoptosis of vascular smooth muscle cells in mice Roberto.

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Presentation transcript:

Aging exacerbates neointimal formation, and increases proliferation and reduces susceptibility to apoptosis of vascular smooth muscle cells in mice Roberto I. Vazquez-Padron, PhD, David Lasko, MD, Sen Li, MD, Louis Louis, MD, Ivo A. Pestana, MD, Manhui Pang, MD, Carlos Liotta, MD, Alessia Fornoni, MD, Abdelouahab Aitouche, PhD, Si M. Pham, MD Journal of Vascular Surgery Volume 40, Issue 6, Pages 1199-1207 (December 2004) DOI: 10.1016/j.jvs.2004.08.034 Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 1 Exaggerated neointima formation in injured aging carotid arteries. A, Representative photomicrographs of cross-sections show neointimal development in aging (left) and young adult (right) mouse carotid arteries subjected to wire injury 4 weeks before harvest (elastica–van Giessen stain; original magnification ×40). B, Intima-media ratio assessed with morphometry in carotid arteries from aging (n = 17) and young (n = 6) mice in response to wire injury. The entire neointimal and medial areas were recorded as pixels from digitized pictures. C, Neointimas in arteries in aging mice (n = 6) had higher cell density than did arteries in young mice (n = 6), as determined at morphometric analysis. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 2 Upregulation of PDGFR-α gene expression in vascular smooth muscle cells (VSMCs) in aging mice. A, Aging increases PDGFR-α in VSMCs as shown on representative complementary DNA arrays. B, Reverse transcriptase polymerase chain reaction confirmation of messenger RNA levels for PDFGR-α in 3 aging and young individual VSMC lines. *P < .05. C, Aging VSMCs respond more readily to PDGF-BB (10 ng/mL) than their younger counterparts do (lower panel). In contrast, aging and young VSMCs proliferate at the same rate in control media (ie, supplemented with 2% fetal calf serum only; upper panel). Cell counts represent the average of at least 3 repeated experiments. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 3 Expression of PDGFR-α in mouse aorta. Abundance of PDGFR-α is present in uninjured aging aortas, as assessed with immunohistochemical staining, in contrast with the barely stained young vessels (right panel). Percent positively stained cells was counted against unstained vascular smooth muscle cells (VSMCs; left panel). *P < .05. VSMC phenotype was ascertained with VSMC α-actin monoclonal antibody. In the negative controls, the primary antibody was substituted with the immunoglobulin G fraction of normal goat serum. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 4 Vascular smooth muscle cells (VSMCs) from aging mice are less susceptible to apoptosis in vitro and in vivo. Aging VSMCs are less susceptible to apoptosis under serum starvation (A) than to exposure to nitric oxide (NO; at 200 μmol/L; (B), as assessed with a Cell Death ELISA kit. Data were normalized for total amount of protein present in each sample. *P < .05). Terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick-end labeled staining (TUNEL) shows more apoptotic cells in the neointima of young compared with aging aortas (C). *P < .01. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 5 Aortic vascular smooth muscle cells from aging mice exhibit greater growth potential in serum-poor (0.1% fetal bovine serum) medium (*P < .05) than those from young mice. Growth increment was expressed as a ratio of the number of cells at a given time (T) and at time zero (To); see Material and methods. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions

Fig 6 No significant differences were noted between aging and young vascular smooth muscle cells with regard to matrix metalloproteinase (MMP-2 and MMP-9) activity at gelatin zymography. Journal of Vascular Surgery 2004 40, 1199-1207DOI: (10.1016/j.jvs.2004.08.034) Copyright © 2004 The Society for Vascular Surgery Terms and Conditions