Leptin, the product of ob gene, is an endocrine hormone that regulates adipose tissue mass. Recently, leptin has been found to generate a growth signal involving a tyrosine kinase-dependent intracellular pathway and promote angiogenic processes via activation of leptin receptor (Ob-R) in endothelial cells. However, it is not clear how leptin functions to promote multi-step processes involved in the neovascularization at the atherosclerotic plaque. We have examined the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) and Ob-R in human atherosclerotic lesions, leptin-mediated angiogenesis in vivo and in vitro. Immunohistochemical analysis of human atherosclerotic aorta revealed an increased expression of Ob-R in the intima of neorevascularized regions and of both MMPs and TIMPs predominantly in the endothelial lining of intimal neovessels and macrophages/foam cells. In the rat corneal angiogenesis assay, leptin elicited a comparable sensitivity of angiogenic activity to those of vascular endothelial growth factor (VEGF). The immunohistological analysis of the leptin-treated rat cornea showed definitive rises in Ob-R, MMPs and TIMPs expression as well as those of VEGF receptor (VEGFR-1). Leptin (10-40 ng/ml) induced proliferation of the human umbilical vein endothelial cells (HUVECs) and elevation of MMP-2, MMP-9, TIMP-1, and TIMP-2 expression in a dose-dependent manner. Leptin also induced increases of MMP-2, MMP-9, TIMP-1, and Up-regulated the human coronary artery smooth muscle cells (HCASMCs). These findings suggest that leptin, a hormone with pluralistic properties including a mitogenic activity on vascular endothelial cells, plays a role in matrix remodeling by regulating the expression of MMPs and TIMPs. Taken together, our findings further provide evidences for leptin's role as an angiogenesis inducer in the normal organ (rat cornea) and in aberrant vasculature under duress like atherosclerosis.
Animal ; Arteriosclerosis/metabolism ; Blotting, Western ; Cell Division ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Growth Factors/metabolism ; Endothelium, Vascular/*cytology/*enzymology ; Enzyme-Linked Immunosorbent Assay ; Fibroblast Growth Factor 2/metabolism ; Immunohistochemistry ; Leptin/*chemistry/metabolism/*physiology ; Lymphokines/metabolism ; Matrix Metalloproteinases/*biosynthesis ; *Neovascularization, Pathologic ; Rats ; Receptor Protein-Tyrosine Kinases/metabolism ; Receptors, Growth Factor/metabolism ; Recombinant Proteins/metabolism ; Support, Non-U.S. Gov't ; Tissue Inhibitor of Metalloproteinases/metabolism ; Umbilical Veins/metabolism ; Up-Regulation