Effect of erythropoietin on angiogenesis with the increased adhesion of platelets to the microvessels in the hind-limb ischemia model in mice.
Erythropoietin (EPO) has been shown to enhance angiogenesis, but its precise mechanisms of enhancement during ischemia are not fully elucidated. We examined the effect of EPO on blood flow recovery from acute hind-limb ischemia induced by ligation of the femoral artery in male C57Bl/6 mice. The density of microvessels with platelet adhesion in ischemic tissues was assessed by intravital microscopy. Treatment with EPO (100 and 1000 IU/kg, i.p.) restored blood flow in a dose-dependent manner and increased plasma levels of soluble-P-selectin, vascular endothelial growth factor (VEGF), and stromal cell-derived factor (SDF-1). Flow cytometric analysis revealed increased P-selectin expression on platelets in EPO-treated mice compared to PBS-treated mice. Intravital microscopic studies showed that EPO increased density of microvessels with platelet adhesion selectively in the ischemic tissues. Neutralizing antibody against P-selectin reduced the density of microvessels with platelet adhesion enhanced with EPO and impaired blood flow recovery with reductions in VEGF and SDF-1 levels. These results suggest that EPO administration enhances recovery from hind-limb ischemia, and platelet adhesion to the microvessels is a key event to enhance the angiogenesis in the ischemic tissues.
Department of Pharmacology, Kitasato University School of Medicine, Kitasato, Sagamihara 228-8555, Japan.
Shintaro
S
Kato
Hideki
H
Amano
Yoshiya
Y
Ito
Koji
K
Eshima
Naoyoshi
N
Aoyama
Hideaki
H
Tamaki
Hiroyuki
H
Sakagami
Yukitoshi
Y
Satoh
Toru
T
Izumi
Masataka
M
Majima
D000818:Animals; D054377:Chemokine CXCL12 / Q000187:drug effects / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004921:Erythropoietin / Q000008:administration & dosage / Q000494:pharmacology*; D005434:Flow Cytometry; D006614:Hindlimb; D006801:Humans; D007511:Ischemia / Q000188:drug therapy* / Q000503:physiopathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D055806:Microvessels / Q000187:drug effects / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D019007:P-Selectin / Q000097:blood / Q000187:drug effects; D010973:Platelet Adhesiveness / Q000187:drug effects*; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A / Q000097:blood / Q000187:drug effects
D054377:Chemokine CXCL12; D019007:P-Selectin; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D004921:Erythropoietin