ABSTRACT
OBJECTIVE: Myeloid lineage cells (MLCs) such as macrophages are known to play a key role in postischemic neovascularization. However, the role of MLC-derived reactive oxygen species in this process and their specific chemical identity remain unknown. METHODS AND RESULTS: Transgenic mice with MLC-specific overexpression of catalase (Tg(Cat-MLC) mice) were created on a C57BL/6 background. Macrophage catalase activity was increased 3.4-fold compared with wild-type mice. After femoral artery ligation, laser Doppler perfusion imaging revealed impaired perfusion recovery in Tg(Cat-MLC) mice. This was associated with fewer collateral vessels, as assessed by microcomputed tomography angiography, and decreased capillary density. Impaired functional recovery of the ischemic limb was also evidenced by a 50% reduction in spontaneous running activity. The deficient neovascularization was associated with a blunted inflammatory response, characterized by decreased macrophage infiltration of ischemic tissues, and lower mRNA levels of inflammatory markers, such as tumor necrosis factor-α, osteopontin, and matrix mettaloproteinase-9. In vitro macrophage migration was impaired in Tg(Cat-MLC) mice, suggesting a role for H(2)O(2) in regulating the ability of macrophages to infiltrate ischemic tissues. CONCLUSIONS: MLC-derived H(2)O(2) plays a key role in promoting neovascularization in response to ischemia and is a necessary factor for the development of ischemia-induced inflammation.
Subject(s)
Capillaries/enzymology , Catalase/biosynthesis , Hydrogen Peroxide/metabolism , Ischemia/enzymology , Muscle, Skeletal/blood supply , Myeloid Cells/enzymology , Neovascularization, Physiologic , Animals , Capillaries/diagnostic imaging , Capillaries/physiopathology , Catalase/genetics , Cell Movement , Cells, Cultured , Collateral Circulation , Disease Models, Animal , Endothelial Cells/metabolism , Femoral Artery/surgery , Genotype , Hindlimb , Humans , Inflammation Mediators/metabolism , Ischemia/genetics , Ischemia/physiopathology , Laser-Doppler Flowmetry , Ligation , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Motor Activity , Neovascularization, Physiologic/genetics , Phenotype , RNA, Messenger/metabolism , Regional Blood Flow , Stem Cells/metabolism , Time Factors , Ultrasonography , Up-Regulation , X-Ray MicrotomographyABSTRACT
OBJECTIVE: Recently, we have shown that shear stress regulates the angiogenic potential of endothelial cells in vitro by an Angiopoietin-2 (Ang2)-dependent mechanism; however its pathophysiological significance in vivo was not clear. We hypothesized that Ang2 plays an important role in blood flow recovery after arterial occlusion in vivo by regulating angiogenesis and arteriogenesis. METHODS AND RESULTS: C57Bl/6J mice underwent femoral artery ligation and were injected with a specific Ang2 inhibitor, L1-10, or vehicle for 10 days. Ang2 mRNA was upregulated at day 2, and Ang2 protein was upregulated at day 2, 5, and 7 in the ligated hindlimb. L1-10 treatment significantly blunted blood flow recovery. L1-10 decreased smooth muscle cell coverage of neovessels without affecting capillary density, suggesting a specific role for Ang2 in arteriogenesis. Mechanistically, L1-10 decreased expression of intercellular and vascular cell adhesion molecules as well as infiltrating monocytes/macrophages in the ischemic tissue. Although L1-10 had no effect on the number of CD11b+ cells (monocytes/macrophages) mobilized in the bone marrow, it maintained elevated numbers of circulating CD11b+ cells in the peripheral blood. CONCLUSIONS: These results suggest that Ang2 induced in ischemic tissue plays a critical role in blood flow recovery by stimulating inflammation and arteriogenesis.
Subject(s)
Angiopoietin-2/metabolism , Arterial Occlusive Diseases/complications , Endothelial Cells/metabolism , Inflammation/metabolism , Ischemia/metabolism , Muscle, Skeletal/blood supply , Neovascularization, Physiologic , Angiopoietin-1/metabolism , Angiopoietin-2/antagonists & inhibitors , Angiopoietin-2/genetics , Animals , Arterial Occlusive Diseases/metabolism , Arterial Occlusive Diseases/physiopathology , CD11b Antigen/blood , Cells, Cultured , Collateral Circulation , Disease Models, Animal , Endothelial Cells/drug effects , Femoral Artery/surgery , Humans , Inflammation/physiopathology , Intercellular Adhesion Molecule-1/metabolism , Ischemia/etiology , Ischemia/physiopathology , Ligation , Male , Mice , Mice, Inbred C57BL , Neovascularization, Physiologic/drug effects , RNA, Messenger/metabolism , Receptor, TIE-2/metabolism , Recombinant Fusion Proteins/pharmacology , Regional Blood Flow , Time Factors , Up-Regulation , Vascular Cell Adhesion Molecule-1/metabolismABSTRACT
The overproduction of hydrogen peroxide is implicated in the development of numerous diseases and there is currently great interest in developing contrast agents that can image hydrogen peroxide in vivo. In this report, we demonstrate that nanoparticles formulated from peroxalate esters and fluorescent dyes can image hydrogen peroxide in vivo with high specificity and sensitivity. The peroxalate nanoparticles image hydrogen peroxide by undergoing a three-component chemiluminescent reaction between hydrogen peroxide, peroxalate esters and fluorescent dyes. The peroxalate nanoparticles have several attractive properties for in vivo imaging, such as tunable wavelength emission (460-630 nm), nanomolar sensitivity for hydrogen peroxide and excellent specificity for hydrogen peroxide over other reactive oxygen species. The peroxalate nanoparticles were capable of imaging hydrogen peroxide in the peritoneal cavity of mice during a lipopolysaccharide-induced inflammatory response. We anticipate numerous applications of peroxalate nanoparticles for in vivo imaging of hydrogen peroxide, given their high specificity and sensitivity and deep-tissue-imaging capability.
