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Assessing Therapeutic Angiogenesis in a Murine Model of Hindlimb Ischemia
Published on: June 8, 2019
Plgf-/-eNos-/- mice show defective angiogenesis associated with increased oxidative stress in response to tissue
Bruna Gigante1, Giulia Morlino, Maria Teresa Gentile
1Institute of Genetics and Biophysics Adriano Buzzati-Traverso, Consiglio Nazionale delle Ricerche, Via P. Castellino, 111, Naples 80131, Italy.
Abstract:
Neo-angiogenesis is a complex phenomenon modulated by the concerted action of several molecular factors. We have generated a congenic line of knockout mice carrying null mutations of both placental growth factor (PlGF) and endothelial nitric oxide synthase (eNOS), two genes that play a pivotal role in the regulation of pathological angiogenesis. In the present study, we describe the phenotype of this new experimental animal model after surgically induced hind-limb ischemia. Plgf-/-, eNos-/-, Plgf-/- eNos-/-, and wild-type C57BL/6J mice were studied. Plgf-/- eNos-/- mice showed the most severe phenotype: self-amputation, and death occurred in up to 47% of the animals studied; in ischemic legs, capillary density was severely reduced; macrophage infiltration and oxidative stress increased as compared to the other groups of animals. These changes were associated with an up-regulation of both inducible NOS (iNOS) expression and vascular endothelial growth factor (VEGF) protein levels in ischemic limbs, and to an increased extent of protein nitration. Our results demonstrate that the deletion of these two genes, Plgf, which acts in synergism with VEGF, and eNos, a downstream mediator of VEGF, determines a significant change in the vascular response to an ischemic stimulus and that oxidative stress within the ischemic tissue represents a crucial factor to maintain tissue homeostasis.
Insights
Mice lacking placental growth factor (PlGF) and endothelial nitric oxide synthase (eNOS) exhibited severe limb ischemia, increased oxidative stress, and mortality. This highlights the critical roles of PlGF and eNOS in vascular response and tissue homeostasis.
Area of Science:
- Vascular Biology
- Molecular Medicine
- Pathophysiology
Background:
- Neo-angiogenesis is regulated by multiple molecular factors.
- Placental growth factor (PlGF) and endothelial nitric oxide synthase (eNOS) are key regulators of pathological angiogenesis.
- Understanding the interplay of these factors is crucial for developing therapeutic strategies.
Purpose of the Study:
- To generate and characterize a congenic mouse model with combined null mutations for PlGF and eNOS.
- To investigate the phenotype of PlGF-/- eNOS-/- mice following surgically induced hind-limb ischemia.
- To elucidate the roles of PlGF and eNOS in the vascular response to ischemic injury and oxidative stress.
Main Methods:
- Generation of a congenic mouse line with combined PlGF and eNOS null mutations.
- Surgically induced hind-limb ischemia model in Plgf-/-, eNos-/-, Plgf-/- eNos-/-, and wild-type mice.
- Assessment of phenotypic severity, capillary density, macrophage infiltration, oxidative stress, iNOS expression, VEGF levels, and protein nitration in ischemic limbs.
Main Results:
- Plgf-/- eNos-/- mice displayed the most severe phenotype, including self-amputation and up to 47% mortality.
- Ischemic legs in these mice showed severely reduced capillary density and increased macrophage infiltration and oxidative stress.
- These alterations were associated with elevated inducible NOS (iNOS) expression, VEGF protein, and protein nitration.
Conclusions:
- Combined deletion of PlGF and eNOS significantly alters the vascular response to ischemic stimuli.
- Oxidative stress in ischemic tissue is a critical factor in maintaining tissue homeostasis.
- This novel animal model provides insights into the complex regulation of angiogenesis and tissue repair.
