Hif-1α Knockdown Reduces Glycolytic Metabolism and Induces Cell Death of Human Synovial Fibroblasts Under Normoxic Conditions.
Hif-1α Knockdown Reduces Glycolytic Metabolism and Induces Cell Death of Human Synovial Fibroblasts Under Normoxic Conditions.
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DOI：
10.1038/s41598-017-03921-4
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发表时间：
2017-06-16
影响因子：
4.6
通讯作者：
Pablos JL
中科院分区：
文献类型：
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作者：
Del Rey MJ;Valín Á;Usategui A;García-Herrero CM;Sánchez-Aragó M;Cuezva JM;Galindo M;Bravo B;Cañete JD;Blanco FJ;Criado G;Pablos JL
糖酵解和HIF-1α活性增加是缺氧或炎症条件下细胞的特征。此外，在正常的O2环境中，糖酵解速率的升高支持关键的细胞机制，如细胞存活。本研究旨在分析缺氧诱导因子-1 α（HIF-1α）对常氧条件下人滑膜成纤维细胞（SF）能量代谢和存活的影响。使用慢病毒载体或小干扰RNA（siRNA）双链体沉默HIF-1α。通过qRT-PCR和western blot对已知HIF-1α靶基因在缺氧条件下的表达分析证实了功能性HIF-1α在常氧SF中的存在，并证实了糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH）即使在常氧条件下也是HIF-1α靶点。HIF-1α沉默在培养的SF中诱导凋亡性细胞死亡，类似地，用糖酵解而不是OXPHOS抑制剂处理诱导SF死亡。最后，通过siRNA靶向的体内HIF-1α显示移植到小鼠气囊中的人SF的活力显著降低。我们的研究结果表明，SF是高度依赖于糖酵解代谢和HIF-1α发挥调节作用，即使在有氧条件下的糖酵解。局部靶向HIF-1α为减少慢性关节炎疾病中SF增生提供了一种可行的策略。
Increased glycolysis and HIF-1α activity are characteristics of cells under hypoxic or inflammatory conditions. Besides, in normal O2 environments, elevated rates of glycolysis support critical cellular mechanisms such as cell survival. The purpose of this study was to analyze the contribution of HIF-1α to the energy metabolism and survival of human synovial fibroblasts (SF) under normoxic conditions. HIF-1α was silenced using lentiviral vectors or small-interfering RNA (siRNA) duplexes. Expression analysis by qRT-PCR and western blot of known HIF-1α target genes in hypoxia demonstrated the presence of functional HIF-1α in normoxic SF and confirmed the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a HIF-1α target even in normoxia. HIF-1α silencing induced apoptotic cell death in cultured SF and, similarly, treatment with glycolytic, but not with OXPHOS inhibitors, induced SF death. Finally, in vivo HIF-1α targeting by siRNA showed a significant reduction in the viability of human SF engrafted into a murine air pouch. Our results demonstrate that SF are highly dependent on glycolytic metabolism and that HIF-1α plays a regulatory role in glycolysis even under aerobic conditions. Local targeting of HIF-1α provides a feasible strategy to reduce SF hyperplasia in chronic arthritic diseases.