Deferoxamine preconditioning to restore impaired HIF-1α-mediated angiogenic mechanisms in adipose-derived stem cells from STZ-induced type 1 diabetic rats.
OBJECTIVE
Both excessive and insufficient angiogenesis are associated with progression of diabetic complications, of which poor angiogenesis is an important feature. Currently, adipose-derived stem cells (ADSCs) are considered to be a promising source to aid therapeutic neovascularization. However, functionality of these cells is impaired by diabetes which can result from a defect in hypoxia-inducible factor-1 (HIF-1), a key mediator involved in neovascularization. In the current study, we sought to explore effectiveness of pharmacological priming with deferoxamine (DFO) as a hypoxia mimetic agent, to restore the compromised angiogenic pathway, with the aid of ADSCs derived from streptozotocin (STZ)-induced type 1 diabetic rats ('diabetic ADSCs').


METHODS
Diabetic ADSCs were treated with DFO and compared to normal and non-treated diabetic ADSCs for expression of HIF-1α, VEGF, FGF-2 and SDF-1, at mRNA and protein levels, using qRT-PCR, western blotting and ELISA assay. Activity of matrix metalloproteinases -2 and -9 were measured using a gelatin zymography assay. Angiogenic potential of conditioned media derived from normal, DFO-treated and non-treated diabetic ADSCs were determined by in vitro (in HUVECs) and in vivo experiments including scratch assay, three-dimensional tube formation testing and surgical wound healing models.


RESULTS
DFO remarkably enhanced expression of noted genes by mRNA and protein levels and restored activity of matrix metalloproteinases -2 and -9. Compromised angiogenic potential of conditioned medium derived from diabetic ADSCs was restored by DFO both in vitro and in vivo experiments.


CONCLUSIONS
DFO preconditioning restored neovascularization potential of ADSCs derived from diabetic rats by affecting the HIF-1α pathway.
Razi Drug Research Center, Department of pharmacology, Iran University of Medical Sciences, Tehran, Iran.
M
M
Mehrabani
Department of Biochemistry, Iran University of Medical Sciences, Tehran, Iran.
M
M
Najafi
Tissue Engineering Group (TEG) & Research, National Orthopedic Centre of Excellence in Research & Learning (NOCERAL), Department of Orthopedics, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
T
T
Kamarul
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
K
K
Mansouri
Department of Pathology, Kerman University of Medical Sciences, Kerman, Iran.
M
M
Iranpour
Department of Biochemistry, Kerman University of Medical Sciences, Kerman, Iran.
M H
MH
Nematollahi
Department of Pharmacology, Tehran University of Medical Sciences, Tehran, Iran.
M
M
Ghazi-Khansari
Razi Drug Research Center, Department of pharmacology, Iran University of Medical Sciences, Tehran, Iran.|Department of Tissue Engineering and regenerative Medicine, School of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
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AM
Sharifi
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000235:genetics / Q000378:metabolism; D003676:Deferoxamine / Q000494:pharmacology*; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000473:pathology; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D008297:Male; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013311:Streptozocin / Q000633:toxicity; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects
C575935:CXCL12 protein, rat; D054377:Chemokine CXCL12; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; D013311:Streptozocin; D003676:Deferoxamine