A 5.0-kD heparin fraction systemically suppresses VEGF165-mediated angiogenesis.

Abstract

The systemic effect of heparin fractions with mean molecular masses of 2.5, 5.0 and 16.4 kD on angiogenesis induced by vascular endothelial growth factor isoform 165 was studied using the truly quantitative rat mesenteric-window angiogenesis assay. The angiogenic treatment with 5 ml of VEGF165 at 480 pM was given intraperitoneally on days 0-4 and heparin fractions were given subcutaneously on days 0-13; animals were sacrificed on day 14. As the overlaps between the molecular mass distributions of the three fractions were relatively small, they essentially represent three different populations of heparin molecules. The doses of the heparins given were equal in terms of weight, but different in terms of the number of molecules and biologic activity. Angiogenesis was assessed in terms of vascularized area (VA), a measurement of microvascular spatial extension, and microvascular length (MVL), a measurement of microvascular density, using technically independent variables and image analysis. The total microvascular length was computed from VA x MVL. Treatment with the 5.0-kD fraction suppressed angiogenesis significantly in statistical terms compared with treatment with 2.5- and 16.4-kD heparins and the saline in controls. Interestingly, the 2.5-kD heparin fraction which was used here has previously been shown statistically significantly to suppress angiogenesis mediated by basic fibroblast growth factor in the same experimental system. Our data thus suggest that the systemic angiosuppressive effect of heparin in different mammalian angiogenic reactions is distinctly related to structural features such as molecular size.