Diabetes mellitus is associated with macro- and micro-angiopathy, leading to increased risk of peripheral ischemia. In the present study, we have characterized the microvascular phenotype at the level of limb muscles and the spontaneous angiogenesis response to surgically-induced unilateral limb ischemia in a murine model of type-2 diabetes, the obese C57BL/KsOlaHsd-Leprdb/db mice (Leprdb/db), and in non-diabetic heterozygous Leprdb/+. Wild type C57BL mice (WT) were used as controls. The basal microvascular phenotype was determined in mice aged 3 or 5 months, while the response to limb ischemia was studied only in 5-month old mice. Moreover, in 5-month old ischemic Leprdb/db and Leprdb/+, we have tested the therapeutic potential of local angiogenesis gene therapy with human tissue kallikrein (hTK) or constitutively-activated Akt kinase (Myr-Akt). We found that in the muscles of 3- or 5-month old Leprdb/db, apoptosis of endothelial cells was enhanced and the densities of capillary and arteriole were reduced. Arterioles of Leprdb/db showed hypertrophic remodelling and, occasionally, lumen occlusion. Following ischemia, Leprdb/db showed a defective reparative angiogenesis in ischemic muscle, delayed blood flow recovery, and worsened clinical outcome as compared with controls. Five-month old Leprdb/+ displayed an increase in endothelial cell apoptosis under basal conditions, while capillary and arteriole densities were normal. Leprdb/+ mounted a proper reparative angiogenesis response to limb ischemia and regained blood flow to the ischemic limb, regularly. Local gene therapy with hTK or Myr-Akt induced angiogenesis in ischemic muscles of Leprdb/+ and Leprdb/db. However, in the Leprdb/db neither gene therapy approach improved the blood flow recovery and the clinical outcome from ischemia. In contrast, either hTK or Myr-Akt gene transfer improved the post-ischemic recovery of Leprdb/+. Type-2 diabetes has a negative impact on the basal microvascular phenotype and severely impairs post-ischemic recovery of limb muscles. Gene therapy-induced stimulation of neovascularization might not suffice as a sole therapeutic strategy to combat type-2 diabetes-related vascular complications. In type-2 diabetic patients, therapeutic angiogenesis may need to be further optimized before being recommended for clinical applications.
|Translated title of the contribution||Type-2 diabetic Lepdb/db mice show a defective microvascular phenotype under basal conditions and an impaired response to angiogenesis gene therapy in the setting of limb ischemia|
|Number of pages||10|
|Journal||Frontiers in Bioscience|
|Publication status||Published - 1 Jan 2007|