13C isotope-based metabolic flux analysis revealing cellular landscape of glucose metabolism in human liver cells exposed to perfluorooctanoic acid

2021 ◽  
Vol 770 ◽  
pp. 145329
Author(s):  
Ruijia Zhang ◽  
Baowei Chen ◽  
Li Lin ◽  
Hui Zhang ◽  
Tiangang Luan
2020 ◽  
Author(s):  
Sheng-nan Ma ◽  
Fei Xie ◽  
Yan Zhang ◽  
Ting-ting Yao ◽  
Meng-yu Liu ◽  
...  

Abstract Background: Our previous study showed that molecular hydrogen could effectively suppress glioblastoma multiforme (GBM) tumor growth via induction of glioblastoma stem cells (GSCs) differentiation. Metabolic reprogramming has been demonstrated to delicately regulate the stemness of cancer stem cells. In the present study, we explored whether metabolic reprogramming is involved in the hydrogen-induced GSCs differentiation. Methods: Immunofluorescence staining was conducted to determine the expression of cell surface markers. Glucose uptake and lactate secretion was determined by spectrophotometric method. Untargeted metabolomics analyses were conducted to investigate the metabolic alteration in GSCs after hydrogen treatment. 13 C-labeled metabolic flux analysis was performed to explore the regulatory effects of molecular hydrogen on the glucose metabolism of GSCs. Results: Immunofluorescence staining showed the up-regulated expression of oligodendroglial markers in hydrogen-treated GSCs. Both glucose uptake and lactate production in GSCs were significantly inhibited by hydrogen treatment. Untargeted metabolomics analyses showed hydrogen-induced promotion of de novo synthesis of nucleotides in GSCs. Metabolic flux analysis showed decreased glucose metabolism in GSCs induced by hydrogen. Conversely, the content of glycerol 3-phosphate, glutamate and glutamine were increased by hydrogen treatment. Conclusions: Results from this study demonstrated that molecular hydrogen could inhibit glucose metabolism and promote de novo synthesis of nucleotides in GSCs, suggesting the involvement of metabolic reprogramming in the hydrogen-induced GSCs differentiation. Our study also provides important new clues to seek the target of molecular hydrogen.


Author(s):  
Reza Afrisham ◽  
Sahar Sadegh-Nejadi ◽  
Reza Meshkani ◽  
Solaleh Emamgholipour ◽  
Molood Bagherieh ◽  
...  

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.


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