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 Sanchez, Nilanjana Maulik

Research output: Contribution to journalArticle

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

Fingerprint

Prolyl Hydroxylases
Hypoxia-Inducible Factor 1
Ischemia
Extremities
Perfusion
Vascular Endothelial Growth Factor A
Peripheral Arterial Disease
Femoral Artery
Immunosorbents
Proteins
Ligation
Fibrosis
Muscle
Assays
Tissue
Imaging techniques
Recovery
Molecules
Lasers
Muscle Cells

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology
  • Medicine(all)

Cite this

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

In: Microvascular Research, Vol. 97, 01.01.2015, p. 181-188.

Research output: Contribution to journalArticle

Rishi, Muhammad T. ; Selvaraju, Vaithinathan ; Thirunavukkarasu, Mahesh ; Shaikh, Inam A. ; Takeda, Kotaro ; Fong, Guo Hua ; Palesty, J. Alexander ; Sanchez, Juan ; Maulik, Nilanjana. / 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. In: Microvascular Research. 2015 ; Vol. 97. pp. 181-188.
@article{9744e59a75ba4b81b305222689293421,
title = "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",
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.",
keywords = "Bcl-2, HIF, Hindlimb ischemia, Prolyl hydroxylase, VEGF",
author = "Rishi, {Muhammad T.} and Vaithinathan Selvaraju and Mahesh Thirunavukkarasu and Shaikh, {Inam A.} and Kotaro Takeda and Fong, {Guo Hua} and Palesty, {J. Alexander} and Juan Sanchez and Nilanjana Maulik",
year = "2015",
month = "1",
day = "1",
doi = "10.1016/j.mvr.2014.10.009",
language = "English (US)",
volume = "97",
pages = "181--188",
journal = "Microvascular Research",
issn = "0026-2862",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - 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

AU - Rishi, Muhammad T.

AU - Selvaraju, Vaithinathan

AU - Thirunavukkarasu, Mahesh

AU - Shaikh, Inam A.

AU - Takeda, Kotaro

AU - Fong, Guo Hua

AU - Palesty, J. Alexander

AU - Sanchez, Juan

AU - Maulik, Nilanjana

PY - 2015/1/1

Y1 - 2015/1/1

N2 - 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.

AB - 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.

KW - Bcl-2

KW - HIF

KW - Hindlimb ischemia

KW - Prolyl hydroxylase

KW - VEGF

UR - http://www.scopus.com/inward/record.url?scp=84918577369&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84918577369&partnerID=8YFLogxK

U2 - 10.1016/j.mvr.2014.10.009

DO - 10.1016/j.mvr.2014.10.009

M3 - Article

C2 - 25446011

AN - SCOPUS:84918577369

VL - 97

SP - 181

EP - 188

JO - Microvascular Research

JF - Microvascular Research

SN - 0026-2862

ER -