Adhesion phenotype manifests an altered metabolic profile favoring glycolysis
Adhesion phenotype manifests an altered metabolic profile favoring glycolysis
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DOI：
10.1016/j.fertnstert.2016.02.009
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发表时间：
2016-06-01
影响因子：
6.7
通讯作者：
Saed, Ghassan M.
中科院分区：
文献类型：
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作者：
Fletcher, Nicole M.;Awonuga, Awoniyi O.;Saed, Ghassan M.
目的：为了确定代谢标志物是否在缺氧暴露和不缺氧暴露的情况下在正常和粘连成纤维细胞中差异表达。设计：前瞻性实验研究。设置：大学研究实验室。患者：从相同患者的正常腹膜和粘连组织中建立的成纤维细胞。干预：在正常和缺氧（20/o 0）条件下对正常腹膜和粘连成纤维细胞进行的体外实验。代谢标志物甘油醛3-磷酸脱氢酶（GAPDH）、葡萄糖转运蛋白1（GLUT 1）、缺氧诱导因子（HIF）-1 α、己糖激酶2（HK 2）、乳糖脱氢酶A（LDHA）的表达，用实时逆转录聚合酶链反应测定细胞内HIF-I α和丙酮酸脱氢酶α 1（PDHA 1）的水平;用ELISA测定三磷酸腺苷（ATP）、HIF-1 α和乳酸水平。与正常腹膜成纤维细胞相比，粘附中GAPDH和HIF-1 α的基线mRNA水平增加，而GLUT 1和PDHA 1降低。在细胞系之间，HK 2或LDHA的基线水平没有变化。在两种细胞系中，缺氧增加了HIF-Ia的蛋白水平和GAPDH、GLUT 1和HK 2的mRNA水平，并降低了PDHA 1的水平。缺氧增加正常腹膜成纤维细胞LDHA mRNA水平。与正常腹膜成纤维细胞相比，粘连中的基线乳酸和ATP水平较低。缺氧后，两种细胞系的乳酸含量均增加，正常成纤维细胞的ATP含量减少。结论：粘附性成纤维细胞的代谢模式发生改变，有利于糖酵解途径，缺氧可进一步改变粘附性成纤维细胞的代谢。在手术过程中靶向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。（Feral Sterile（R）2016;105：1628-37。(C)2016年美国生殖医学会（American Society for Reproductive Medicine）
Objective: To determine whether metabolic markers are differentially expressed in normal and adhesion fibroblasts with and without hypoxia exposure.Design: Prospective experimental study.Setting: 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 (20/o 0,) conditions.Main Outcome Measure(s): Expression of metabolic markers, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), glucose transporter 1 (GLUT 1), hypoxia inducible factor (HIF)-1a, 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- la, and lactate levels were assessed with ELISAs.Result(s): Baseline mRNA levels of GAPDH and HIF- la 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- la 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. (Feral Sterile(R) 2016;105:1628-37. (C) 2016 by American Society for Reproductive Medicine.)