Uncoupling oxidative phosphorylation with 2,4-dinitrophenol promotes development of the adhesion phenotype.
OBJECTIVE
To determine the effect of uncoupling oxidative phosphorylation with 2,4-dinitrophenol (DNP) on adhesion phenotype development.


METHODS
Prospective experimental study.


METHODS
Academic medical center.


METHODS
Women undergoing laparotomy for pelvic pain from whom normal peritoneum and adhesions were excised to create primary cultures of normal peritoneal and adhesion fibroblasts.


METHODS
Treatment of normal peritoneal and adhesion fibroblasts isolated from the same patient(s) with or without 0.2 mM DNP for 24 hours.


METHODS
Evaluation of adhesion phenotype markers type I collagen, vascular endothelial growth factor (VEGF), and hypoxia-inducible factor (HIF)-1α.


RESULTS
In agreement with prior findings, adhesion fibroblasts exhibited significantly higher basal levels of type I collagen, VEGF, and HIF-1α compared with normal peritoneal fibroblasts. Treatment of normal peritoneal fibroblasts with DNP resulted in significant increases in type I collagen (10.2 ± 1.4 vs. 18.4 ± 1.9 fg/μg RNA) and VEGF (8.2 ± 1.1 vs. 13.7 ± 0.4 fg/μg RNA) over baseline. HIF-1α levels did not increase when normal peritoneal fibroblasts were treated with DNP.


CONCLUSIONS
The adhesion phenotype, which is normally expressed in response to hypoxia, is reproduced in a normoxic environment by uncoupling oxidative phosphorylation with DNP, as evidenced by an increase in type I collagen and VEGF. Acquisition of the adhesion phenotype was via a mechanism distinct from up-regulation of HIF-1α. These observations are consistent with the hypothesis that the adhesion phenotype represents a state of intracellular metabolic depletion.
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, The CS Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. vshavell@med.wayne.edu
Valerie I
VI
Shavell
Nicole M
NM
Fletcher
Zhong L
ZL
Jiang
Ghassan M
GM
Saed
Michael P
MP
Diamond
D019297:2,4-Dinitrophenol / Q000494:pharmacology*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D010085:Oxidative Phosphorylation / Q000187:drug effects*; D017699:Pelvic Pain / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D000267:Tissue Adhesions; D014475:Uncoupling Agents / Q000494:pharmacology*; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism
D024042:Collagen Type I; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012333:RNA, Messenger; D014475:Uncoupling Agents; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D019297:2,4-Dinitrophenol