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Biochimica et biophysica acta, General subjects v.1861 no.3, 2017년, pp.673 - 682   SCI SCIE
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Hypoxia mimetic induces lipid accumulation through mitochondrial dysfunction and stimulates autophagy in murine preadipocyte cell line

Marques, Ana Patrícia (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ; Rosmaninho-Salgado, Joana (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ; Estrada, Marta (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ; Cortez, Vera (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ; Nobre, Rui Jorge (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ; Cavadas, Cláudia (CNC - Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal ) ;
  • 초록  

    Abstract Background Hypoxia occurs within adipose tissue of obese human and mice. However, its role in adipose tissue regulation is still controversial. Methods We used murine preadipocyte 3T3-L1 cells and hypoxia was induced by using hypoxia mimetic agents, as CoCl 2 . To study adipocyte differentiation, we evaluated the adipocyte markers (PPARγ, C/EBPα and aP2), and a preadipocyte marker (pref-1) by qPCR, western blotting and immunofluorescence. Lipid accumulation was evaluated by Oil red-O assay and perilipin levels by western blotting and immunofluorescence. The effect of CoCl 2 in microRNA, miR-27a and miR-27b, levels was evaluated by qPCR. We also assessed the mitochondrial membrane potential and reactive oxygen species (ROS), superoxide and ATP production. The effect of hypoxia mimetic in autophagy was determined by LC3B and p62 level evaluation by western blotting. Results Our results show that the hypoxia mimetic cobalt chloride increases lipid accumulation with no expression of PPARγ2. Furthermore, using qPCR we observed that the hypoxia mimetic increases microRNAs miR-27a and miR-27b, which are known to block PPARγ2 expression. In contrast, cobalt chloride induces mitochondrial dysfunction, and increases ROS production and autophagy. Moreover, an antioxidant agent, glutathione, prevents lipid accumulation induced by hypoxia mimetic indicating that ROS are responsible for hypoxia-induced lipid accumulation. Conclusions All these results taken together suggest that hypoxia mimetic blocks differentiation and induces autophagy. Hypoxia mimetic also induces lipid accumulation through mitochondrial dysfunction and ROS accumulation. General significance This study highlights the importance of adipocyte response to hypoxia, which might impair adipocyte metabolism and compromise adipose tissue function. Highlights Hypoxia mimetic inhibits adipocyte differentiation and increases miR-27a and b. Hypoxia mimetic causes mitochondrial dysfunction and an increase in reactive oxygen species. Hypoxia mimetic induces lipid accumulation through reactive oxygen species increase. Hypoxia mimetic induces autophagy activation. Graphical abstract [DISPLAY OMISSION]


  • 주제어

    Hypoxia .   Adipogenesis .   Mitochondrial dysfunction .   Reactive oxygen species .   Autophagy.  

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