Journal: Frontiers in Physiology

Article Title: Soluble Fms-Like Tyrosine Kinase-1 Alters Cellular Metabolism and Mitochondrial Bioenergetics in Preeclampsia

doi: 10.3389/fphys.2018.00083

Figure Lengend Snippet: Role of VEGF in mitochondrial bioenergetics in preeclampsia. ECs and HTR-8/SVneo cells were treated with 2% serum from NOR, CTL, and PE women with and without exogenous VEGF (20 ng/mL) for 24 h. (A) Maximal respiration (OCR after FCCP administration) and spare respiratory capacity (Difference between basal and maximal OCR) in endothelial cells, (B) Basal and maximal (Max) RCR, (C,D) are the same experiments as in (A,B) , respectively, but evaluated in HTR-8/SVneo cells. Data are presented as means ± SEM. ( n = 5). In (A,C) : * P < 0.05, vs. NOR, # P < 0.05, vs. cells exposed to PE serum alone. In (B,D) : * P < 0.05, vs. NOR maximal RCR, # P < 0.05, vs. PE maximal RCR. ANOVA ( Bonferroni's post hoc test ).

Article Snippet: Bovine aortic endothelial cells (ECs) (passages 5–8) were obtained from Cell Applications, Inc. (San Diego, CA, USA).

Techniques:

Journal: Frontiers in Physiology

Article Title: Soluble Fms-Like Tyrosine Kinase-1 Alters Cellular Metabolism and Mitochondrial Bioenergetics in Preeclampsia

doi: 10.3389/fphys.2018.00083

Figure Lengend Snippet: sFlt-1 induces a metabolic phenotype switch to glycolysis in ECs but not in trophoblasts. ECs and HTR-8/SVneo cells were exposed to 0, 10, 25, and 50 ng/mL of rh-sFlt-1 for 24 h. (A) Oxygen consumption rates (OCR), maximal respiration (OCR after FCCP administration) and spare respiratory capacity (Difference between basal and maximal OCR), (B) Energetic phenotype map, basal and maximal OCR and extracellular acidification rates (ECAR), (C) ECAR determinations of glycolysis rate and glycolytic reserve, evaluated in endothelial cells. (D-F) , are the same experiments as in (A–C) , respectively, but evaluated in HTR-8/SVneo cells. Data is presented as means ± SEM. ( n = 5), * P < 0.05, ** P < 0.01, *** P < 0.001 vs. untreated controls, In (A,D) : Student T -test. In (C,F) : ANOVA ( Bonferroni's post hoc test ).

Article Snippet: Bovine aortic endothelial cells (ECs) (passages 5–8) were obtained from Cell Applications, Inc. (San Diego, CA, USA).

Techniques:

Journal: Frontiers in Physiology

Article Title: Soluble Fms-Like Tyrosine Kinase-1 Alters Cellular Metabolism and Mitochondrial Bioenergetics in Preeclampsia

doi: 10.3389/fphys.2018.00083

Figure Lengend Snippet: sFlt-1 acts as a mitochondrial bioenergetics disruptor in preeclampsia. (A) Morphological changes in endothelial cells (ECs) and trophoblasts cultured in glucose and galactose media (40X). (B) Cell viability of ECs and (C) trophoblasts cultured in glucose and galactose media and exposed to exogenous 50 ng/mL of sFlt-1 during 24 h. Scale: 100 μm. Data are presented as means ± SEM. ( n = 3), ** P < 0.01, *** P < 0.001, vs. galactose exposed cells. # P < 0.05, vs. glucose-exposed cells. Student T -test.

Article Snippet: Bovine aortic endothelial cells (ECs) (passages 5–8) were obtained from Cell Applications, Inc. (San Diego, CA, USA).

Techniques: Cell Culture

Journal: Frontiers in Physiology

Article Title: Soluble Fms-Like Tyrosine Kinase-1 Alters Cellular Metabolism and Mitochondrial Bioenergetics in Preeclampsia

doi: 10.3389/fphys.2018.00083

Figure Lengend Snippet: sFlt-1 induces mitochondrial dysfunction in vitro . (A) Mitochondrial ROS (mtROS) determinations by fluorescent microscopy using MitoSOX Red fluorescent probe demonstrate that sFlt-1 significantly induced mtROS formation in endothelial cells (ECs), while these effects were not observed in (B) trophoblasts. (C) sFlt-1 dissipated the mitochondrial membrane potential (Ψ m ) measured by fluorescent microscopy using JC-1 fluorescent probe in ECs, but not in (D) trophoblasts. Cells were exposed to sFlt-1 (50 ng/mL) for 24 h. Data is presented as means ± SEM. ( n = 3), * P < 0.05 vs. untreated controls. Student T -test.

Article Snippet: Bovine aortic endothelial cells (ECs) (passages 5–8) were obtained from Cell Applications, Inc. (San Diego, CA, USA).

Techniques: In Vitro, Microscopy, Membrane

Journal: Frontiers in Physiology

Article Title: Soluble Fms-Like Tyrosine Kinase-1 Alters Cellular Metabolism and Mitochondrial Bioenergetics in Preeclampsia

doi: 10.3389/fphys.2018.00083

Figure Lengend Snippet: Schematic view of the effects of sFlt-1 dysregulation over cellular metabolism and bioenergetics in PE. Dysregulated VEGF signaling due to up-regulation of sFlt-1 levels in preeclampsia leads to reduced activation of VEGF receptors Flt-1 and Flk-1/KDR, respectively. Effects of dysregulated VEGF bioavailability affect mitochondrial oxygen consumption (OCR), inducing a metabolic phenotype switch enhancing glycolytic response (ECAR) in endothelial cells, but not in trophoblasts. sFlt-1 due to loss of mitochondrial bioenergetics increase oxidative stress in mitochondria (mtROS). Together, these events lead to mitochondrial dysfunction, that would result in vascular dysfunction and the onset of preeclampsia.

Article Snippet: Bovine aortic endothelial cells (ECs) (passages 5–8) were obtained from Cell Applications, Inc. (San Diego, CA, USA).

Techniques: Activation Assay