Deletion of prolyl hydroxylase domain proteins (PHD1, PHD3) stabilizes hypoxia inducible factor-1 alpha, promotes neovascularization, and improves perfusion in a murine model of hind-limb ischemia

Muhammad T. Rishi, Vaithinathan Selvaraju, Mahesh Thirunavukkarasu, Inam A. Shaikh, Kotaro Takeda, Guo Hua Fong, J. Alexander Palesty, Juan A. Sanchez, Nilanjana Maulik

Research output: Contribution to journalArticlepeer-review

Abstract

Background: There is an emerging focus on investigating innovative therapeutic molecules that can potentially augment neovascularization in order to treat peripheral arterial disease (PAD). Although prolyl hydroxylase domain proteins 1 and 3 (PHD1 and PHD3) may modulate angiogenesis via regulation of hypoxia inducible factor-1α (HIF-1α), there has been no study directly addressing their roles in ischemia-induced vascular growth. We hypothesize that PHD1-/- or PHD3-/- deficiency might promote angiogenesis in the murine hind-limb ischemia (HLI) model. Study design: Wild type (WT), PHD1-/- and PHD3-/- male mice aged 8-12weeks underwent right femoral artery ligation. Post-procedurally, motor function assessment and laser Doppler imaging were periodically performed. The mice were euthanized after 28days and muscles were harvested. Immunohistochemical analysis was performed to determine the extent of angiogenesis by measuring capillary and arteriolar density. VEGF expression was quantified by enzyme-linked immunosorbent assay (ELISA). Bcl-2 and HIF-1α were analyzed by immunofluorescence. Fibrosis was measured by picrosirius red staining. Results: PHD1-/- and PHD3-/- mice showed significantly improved recovery of perfusion and motor function score when compared to WT after femoral artery ligation. These mice also exhibited increased capillary and arteriolar density, capillary/myocyte ratio along with decreased fibrosis compared to WT. VEGF, Bcl-2 and HIF-1α expression increased in PHD1-/- and PHD3-/- mice compared to WT. Conclusions: Taken together these results suggest that PHD1 and PHD3 deletions promote angiogenesis in ischemia-injured tissue, and may present a promising therapeutic strategy in treating PAD.

Original languageEnglish (US)
Pages (from-to)181-188
Number of pages8
JournalMicrovascular Research
Volume97
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

Keywords

  • Bcl-2
  • HIF
  • Hindlimb ischemia
  • Prolyl hydroxylase
  • VEGF

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology

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