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Topical prolyl hydroxylase domain‐2 silencing improves diabetic murine wound closure
Author(s) -
Wetterau Meredith,
George Finny,
Weinstein Andrew,
Nguyen Phuong D.,
Tutela John Paul,
Knobel Denis,
Cohen, BA Oriana,
Warren Stephen M.,
Saadeh Pierre B.
Publication year - 2011
Publication title -
wound repair and regeneration
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.847
H-Index - 109
eISSN - 1524-475X
pISSN - 1067-1927
DOI - 10.1111/j.1524-475x.2011.00697.x
Subject(s) - angiogenesis , small interfering rna , hypoxia inducible factors , gene silencing , vascular endothelial growth factor , wound healing , cancer research , cd31 , chemistry , microbiology and biotechnology , endocrinology , medicine , biology , rna , immunology , biochemistry , vegf receptors , gene
ABSTRACT Prolyl hydroxylase domain 2 (PHD2) has been implicated in several pathways of cell signaling, most notably in its regulation of hypoxia‐inducible factor (HIF)‐1α stability. In normoxia, PHD2 hydroxylates proline residues on HIF‐1α, rendering it inactive. However, in hypoxia, PHD2 is inactive, HIF‐1α is stabilized and downstream effectors such as vascular endothelial growth factor and fibroblast growth factor‐2 are produced to promote angiogenesis. In the present study we utilize RNA interference to PHD2 to promote therapeutic angiogenesis in a diabetic wound model, presumably by the stabilization of HIF‐1α. Stented wounds were created on the dorsum of diabetic Lepr db/db mice. Mice were treated with PHD2 small interfering RNA (siRNA) or nonsense siRNA. Wounds were measured photometrically on days 0–28. Wounds were harvested for histology, protein, and RNA analysis. Diabetic wounds treated with siRNA closed within 21±1.2 days; sham‐treated closed in 28±1.5 days. By day 7, Western blot revealed near complete suppression of PHD protein and corresponding increased HIF‐1α. Angiogenic mediators vascular endothelial growth factor and fibroblast growth factor‐2 were elevated, corresponding to increased CD31 staining in the treated groups. siRNA‐mediated silencing of PHD2 increases HIF‐1α and several mediators of angiogenesis. This corresponded to improved time to closure in diabetic wounds compared with sham‐treated wounds. These findings suggest that impaired wound healing in diabetes can be ameliorated with therapeutic angiogenesis.