Blockade of advanced glycation end-product formation restores ischemia-induced angiogenesis in diabetic mice

doi:10.1073/pnas.1236929100

Blockade of advanced glycation end-product formation restores ischemia-induced angiogenesis in diabetic mice

^*Institut National de la Santé et de la Recherche Médicale (INSERM) U541, Hôpital Lariboisière, Institut Federatif de Recherche Circulation-Paris 7, Université Paris 7-Denis Diderot, 75475 Paris Cedex 10, France; ^†Cardiovascular Research Institute Maastricht, Maastricht, 6202 AZ, The Netherlands; and ^‡INSERM U76 and Institut National de la Transfusion Sanguine (INTS), 75739 Paris Cedex 15, France

Edited by Anthony Cerami, The Kenneth S. Warren Institute, Kitchawan, NY (received for review November 14, 2002)

Abstract

We hypothesized that formation of advanced glycation end products (AGEs) associated with diabetes reduces matrix degradation by metalloproteinases (MMPs) and contributes to the impairment of ischemia-induced angiogenesis. Mice were treated or not with streptozotocin (40 mg/kg) and streptozotocin plus aminoguanidine (AGEs formation blocker, 50 mg/kg). After 8 weeks of treatment, hindlimb ischemia was induced by right femoral artery ligature. Plasma AGE levels were strongly elevated in diabetic mice when compared with control mice (579 ± 21 versus 47 ± 4 pmol/ml, respectively; P < 0.01). Treatment with aminoguanidine reduced AGE plasma levels when compared with untreated diabetic mice (P < 0.001). After 28 days of ischemia, ischemic/nonischemic leg angiographic score, capillary density, and laser Doppler skin-perfusion ratios were 1.4-, 1.5-, and 1.4-fold decreased in diabetic mice in reference to controls (P < 0.01). Treatment with aminoguanidine completely normalized ischemia-induced angiogenesis in diabetic mice. We next analyzed the role of proteolysis in AGE formation-induced hampered neovascularization process. After 3 days of ischemia, MMP-2 activity and MMP-3 and MMP-13 protein levels were increased in untreated and aminoguanidine-treated diabetic mice when compared with controls (P < 0.05). Despite this activation of the MMP pathway, collagenolysis was decreased in untreated diabetic mice. Conversely, treatment of diabetic mice with aminoguanidine restored collagenolysis toward levels found in control mice. In conclusion, blockade of AGE formation by aminoguanidine normalizes impaired ischemia-induced angiogenesis in diabetic mice. This effect is probably mediated by restoration of matrix degradation processes that are disturbed as a result of AGE accumulation.

Footnotes

↵ § To whom correspondence should be addressed at: Institut National de la Santé et de la Recherche Médicale U541, Hôpital Lariboisière, 41 Bd de la Chapelle, 75475 Paris Cedex 10, France. E-mail: levy{at}infobiogen.fr.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: AGE, advanced glycation end product; MMP, metalloproteinase; STZ, streptozotocin; TIMP, tissue inhibitor of metalloproteinase; VEGF, vascular endothelial growth factor.