YQ, Ou JS. Endothelium-derived microparticles inhibit angiogenesis in the heart and enhance the inhibitory effects of hypercholesterolemia on angiogenesis. Am J Physiol Endocrinol Metab 300: E661-E668, 2011. First published January 18, 2011; doi:10.1152/ajpendo.00611.2010.-Therapeutic angiogenesis remains unsuccessful in coronary artery disease. It is known that plasma endothelium-derived microparticles (EMPs) are increased in coronary artery disease and that hypercholesterolemia can inhibit angiogenesis. We evaluated the relationship between EMPs and hypercholesterolemia in the impairment of angiogenesis. EMPs isolated from human umbilical vein endothelial cells were injected into low-density lipoprotein receptor-null (LDLr Ϫ/Ϫ ) mice fed a Western diet for 2 wk and C57BL6 mice for 6 h or were directly added to the tissue culture media. Hearts isolated from mice were sectioned and cultured, and endothelial tube formation was measured. The expression and phosphorylation of endothelial NO synthase (eNOS) and the generation of NO in the hearts were determined. Angiogenesis was inhibited by pathophysiological concentrations of EMPs but not physiological concentrations of EMPs in hearts from C57BL6 mice. However, angiogenesis was inhibited by EMPs at both physiological and pathophysiological concentrations of EMPs in hearts from hypercholesterolemic LDLr Ϫ/Ϫ mice. Pathophysiological concentrations of EMPs decreased eNOS phosphorylation at Ser 1177 and NO generation without altering eNOS expression in hearts from C57BL6 mice. Both physiological and pathophysiological concentrations of EMPs decreased not only eNOS phosphorylation at Ser 1177 and NO generation, but eNOS expression in hypercholesterolemic hearts from LDLr Ϫ/Ϫ mice. These data demonstrated that pathophysiological concentrations of EMPs could inhibit angiogenesis in hearts by decreasing eNOS activity. EMPs and hypercholesterolemia mutually enhanced their inhibitory effect of angiogenesis by inducing eNOS dysfunction. Our findings suggest a novel mechanism by which hypercholesterolemia impairs angiogenesis. cardiovascular diseases; endothelial nitric oxide synthase; nitric oxide ENDOTHELIUM-DERIVED MICROPARTICLES (EMPs) are subcellular fragments of the endothelial cell lipid bilayer (7,39). EMPs are present at relatively low concentrations under normal physiological conditions and, upon activation, are released from endothelial cells (6,7,26,39). Elevated plasma concentrations of EMPs have been reported in many cardiovascular diseases, such as coronary artery disease (20,30,51), acute coronary syndrome (2,3,25), and hypertension (16,29,36,48). Indeed, an increase in the concentration of EMPs has been suggested as a marker of endothelial dysfunction (15).Angiogenesis has been identified as a potential therapy for myocardial ischemia due to coronary artery disease. Many studies in animal models have successfully increased angiogenesis in the heart (13, 38, 43). However, clinical trials have failed to demonstrate that therapeutic angiogenesis is as effective ...