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.
Fertil Steril. 2012 Mar;97(3):729-33. doi: 10.1016/j.fertnstert.2011.12.009. Epub 2011 Dec 24.
To determine the effect of uncoupling oxidative phosphorylation with 2,4-dinitrophenol (DNP) on adhesion phenotype development.
Prospective experimental study.
Academic medical center.
PATIENT(S): Women undergoing laparotomy for pelvic pain from whom normal peritoneum and adhesions were excised to create primary cultures of normal peritoneal and adhesion fibroblasts.
INTERVENTION(S): Treatment of normal peritoneal and adhesion fibroblasts isolated from the same patient(s) with or without 0.2 mM DNP for 24 hours.
MAIN OUTCOME MEASURE(S): Evaluation of adhesion phenotype markers type I collagen, vascular endothelial growth factor (VEGF), and hypoxia-inducible factor (HIF)-1α.
RESULT(S): 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.
CONCLUSION(S): 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.
确定解偶联氧化磷酸化用 2,4-二硝基苯酚（DNP）对黏附表型发育的影响。
前瞻性实验研究。
学术医疗中心。
因盆腔痛而行剖腹术的女性，从中切除正常腹膜和粘连组织，以创建正常腹膜和粘连成纤维细胞的原代培养物。
用或不用 0.2mM DNP 处理来自同一患者的正常腹膜和粘连成纤维细胞 24 小时。
评估黏附表型标志物 I 型胶原、血管内皮生长因子（VEGF）和缺氧诱导因子（HIF）-1α。
与先前的发现一致，黏附成纤维细胞与正常腹膜成纤维细胞相比，基底水平的 I 型胶原、VEGF 和 HIF-1α 显著升高。用 DNP 处理正常腹膜成纤维细胞导致 I 型胶原（10.2±1.4 对 18.4±1.9fg/μg RNA）和 VEGF（8.2±1.1 对 13.7±0.4fg/μg RNA）明显高于基础水平。用 DNP 处理正常腹膜成纤维细胞时，HIF-1α 水平没有增加。
黏附表型通常在缺氧时表达，用 DNP 解偶联氧化磷酸化可在常氧环境中重现，表现为 I 型胶原和 VEGF 的增加。黏附表型的获得是通过一种与 HIF-1α 上调不同的机制。这些观察结果与这样的假设一致，即黏附表型代表细胞内代谢耗竭的状态。
