Fletcher Nicole M, Awonuga Awoniyi O, Abusamaan Mohammed S, Saed Mohammed G, Diamond Michael P, Saed Ghassan M
Department of Obstetrics and Gynecology, C.S. Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan.
Department of Obstetrics and Gynecology, Georgia Regents University, Augusta, Georgia.
Fertil Steril. 2016 Jun;105(6):1628-1637.e1. doi: 10.1016/j.fertnstert.2016.02.009. Epub 2016 Feb 23.
To determine whether metabolic markers are differentially expressed in normal and adhesion fibroblasts with and without hypoxia exposure.
Prospective experimental study.
University research laboratory.
PATIENT(S): Fibroblasts established from normal peritoneum and adhesion tissues from the same patients.
INTERVENTION(S): In vitro experiments on normal peritoneal and adhesion fibroblasts under normal and hypoxic (2% O2) conditions.
MAIN OUTCOME MEASURE(S): Expression of metabolic markers, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), glucose transporter 1 (GLUT1), hypoxia inducible factor (HIF)-1α, hexokinase 2 (HK2), lactose dehydrogenase A (LDHA), and pyruvate dehydrogenase alpha 1 (PDHA1) were measured using real-time reverse transcription polymerase chain reaction; adenosine triphosphate (ATP), HIF-1α, and lactate levels were assessed with ELISAs.
RESULT(S): Baseline mRNA levels of GAPDH and HIF-1α were increased, while GLUT1 and PDHA1 were decreased in adhesion as compared with in normal peritoneal fibroblasts. There was no change in baseline levels of HK2 or LDHA between the cell lines. Hypoxia increased protein levels of HIF-1α and mRNA levels of GAPDH, GLUT1, and HK2 and decreased levels of PDHA1 in both cell lines. Hypoxia increased LDHA mRNA levels in normal peritoneal fibroblasts. Baseline levels of lactate and ATP were lower in adhesion as compared with in normal peritoneal fibroblasts. In response to hypoxia, there was an increase in lactate in both cell lines and a decrease in ATP in normal fibroblasts.
CONCLUSION(S): Adhesion fibroblasts manifested an altered metabolic profile, which favors the glycolytic pathway, and is further altered by hypoxia. Targeting these specific metabolic markers during surgery can be an important therapeutic intervention minimizing the development of postoperative adhesions.
确定代谢标志物在有无缺氧暴露的正常和粘连成纤维细胞中是否差异表达。
前瞻性实验研究。
大学研究实验室。
从同一患者的正常腹膜和粘连组织中分离出的成纤维细胞。
在正常和缺氧（2%氧气）条件下对正常腹膜和成纤维细胞进行体外实验。
使用实时逆转录聚合酶链反应测量代谢标志物甘油醛-3-磷酸脱氢酶（GAPDH）、葡萄糖转运蛋白1（GLUT1）、缺氧诱导因子（HIF）-1α、己糖激酶2（HK2）、乳酸脱氢酶A（LDHA）和丙酮酸脱氢酶α1（PDHA1）的表达；用酶联免疫吸附测定法评估三磷酸腺苷（ATP）、HIF-1α和乳酸水平。
与正常腹膜成纤维细胞相比，粘连成纤维细胞中GAPDH和HIF-1α的基线mRNA水平升高，而GLUT1和PDHA1降低。两种细胞系中HK2或LDHA的基线水平没有变化。缺氧增加了两种细胞系中HIF-1α的蛋白水平以及GAPDH、GLUT1和HK2的mRNA水平，并降低了PDHA1的水平。缺氧增加了正常腹膜成纤维细胞中LDHA的mRNA水平。与正常腹膜成纤维细胞相比，粘连成纤维细胞中乳酸和ATP的基线水平较低。对缺氧的反应是，两种细胞系中的乳酸均增加，正常成纤维细胞中的ATP减少。
粘连成纤维细胞表现出代谢谱改变，有利于糖酵解途径，并且在缺氧时进一步改变。手术期间针对这些特定的代谢标志物可能是一种重要的治疗干预措施，可最大限度地减少术后粘连的发生。
