title: Alterations in endothelial cell proliferation and apoptosis contribute to vascular remodeling following hind-limb ischemia in rabbits.
abstract: Hind-limb ischemia is a potent stimulus for angiogenesis. However, capillary density does not change in tibialis anterior muscle (TA) following hind-limb ischemia, despite increases in angiogenic growth factors. The objective of this study was to determine whether changes in proliferation and apoptosis occurred in the same muscle. In total, 19 New Zealand white rabbits underwent femoral artery ligation and excision and the ischemic and contra-lateral (control) TA muscles were harvested after 1 (n = 7), 5 (n = 7) and 21 (n= 5). Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was used to detect apoptosis and double staining was used to identify the apoptotic cell types. Proliferation was assessed by immunohistochemistry for proliferating cell nuclear antigen (PCNA) and [3H]thymidine incorporation, in vitro. TUNEL positive nuclei were greater in ischemic than control muscle at 1 day (1.83 +/- 0.70% vs 1.03 +/- 0.20%), 5 days (2.13 +/- 0.50% vs 1.21 +/- 0.42%) and at 21 days the difference was statistically significant (3.42 +/- 0.80% vs 0.96 +/- 0.40%, p < 0.01). The majority of TUNEL positive nuclei were endothelial (Tie2 positive) cells. The number of PCNA positive cells in ischemic versus control muscle was similar at 1 day (0.71 +/- 0.20% vs 0.53 +/- 0.20%) and 5 days (1.28 +/- 0.30% vs 0.77 +/- 0.30%), but was significantly (p < 0.05) reduced in ischemic muscle at 21 days (0.18 +/- 0.20% vs 1.35 +/- 0.30%) with no difference in [3H]thymidine incorporation. Directionally opposite changes in endothelial cell proliferation and apoptosis occur in TA muscle following hind-limb ischemia. Modulating apoptosis in ischemic skeletal muscle may present a novel therapeutic target in peripheral arterial disease.
authors.affiliation: Department of Medicine, Durham VA and Duke University Medical Center, North Carolina 27710, USA.
authors.forename: Qunsheng
authors.initials: Q
authors.lastname: Dai
authors.forename: Michael A
authors.initials: MA
authors.lastname: Thompson
authors.forename: Anne M
authors.initials: AM
authors.lastname: Pippen
authors.forename: Hunter
authors.initials: H
authors.lastname: Cherwek
authors.forename: Doris A
authors.initials: DA
authors.lastname: Taylor
authors.forename: Brian H
authors.initials: BH
authors.lastname: Annex
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism / Q000502:physiology; D004195:Disease Models, Animal; D016228:Endothelial Growth Factors / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology*; D005121:Extremities / Q000098:blood supply*; D005260:Female; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007511:Ischemia / Q000378:metabolism / Q000503:physiopathology*; D008222:Lymphokines / Q000378:metabolism; D008955:Models, Cardiovascular; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism / Q000503:physiopathology; D032390:Myoblasts, Smooth Muscle / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; D018809:Proliferating Cell Nuclear Antigen; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors