Negmadjanov 2015 PLOS ONE

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Negmadjanov U, Godic Z, Rizvi F, Emelyanova L, Ross G, Richards J, Holmuhamedov EL, Jahangir A (2015) TGF-Ξ²1-mediated differentiation of fibroblasts is associated with increased mitochondrial content and cellular respiration. PLOS ONE 10:e0123046. doi: 10.1371/journal.pone.0123046.

Β» PMID: 25849590 Open Access

Negmadjanov U, Godic Z, Rizvi F, Emelyanova L, Ross G, Richards J, Holmuhamedov EL, Jahangir A (2015) PLOS ONE

Abstract: Objectivs: Cytokine-dependent activation of fibroblasts to myofibroblasts, a key event in fibrosis, is accompanied by phenotypic changes with increased secretory and contractile properties dependent on increased energy utilization, yet changes in the energetic profile of these cells are not fully described. We hypothesize that the TGF-Ξ²1-mediated transformation of myofibroblasts is associated with an increase in mitochondrial content and function when compared to naive fibroblasts.

Methods: Cultured NIH/3T3 mouse fibroblasts treated with TGF-Ξ²1, a profibrotic cytokine, or vehicle were assessed for transformation to myofibroblasts (appearance of Ξ±-smooth muscle actin [Ξ±-SMA] stress fibers) and associated changes in mitochondrial content and functions using laser confocal microscopy, Seahorse respirometry, multi-well plate reader and biochemical protocols. Expression of mitochondrial-specific proteins was determined using western blotting, and the mitochondrial DNA quantified using Mitochondrial DNA isolation kit.

Results: Treatment with TGF-Ξ²1 (5 ng/mL) induced transformation of naive fibroblasts into myofibroblasts with a threefold increase in the expression of Ξ±-SMA (6.85 Β± 0.27 RU) compared to cells not treated with TGF-Ξ²1 (2.52 Β± 0.11 RU). TGF-Ξ²1 exposure increased the number of mitochondria in the cells, as monitored by membrane potential sensitive dye tetramethylrhodamine, and expression of mitochondria-specific proteins; voltage-dependent anion channels (0.54 Β± 0.05 vs. 0.23 Β± 0.05 RU) and adenine nucleotide transporter (0.61 Β± 0.11 vs. 0.22 Β± 0.05 RU), as well as mitochondrial DNA content (530 Β± 12 ΞΌg DNA/106 cells vs. 307 Β± 9 ΞΌg DNA/106 cells in control). TGF-Ξ²1 treatment was associated with an increase in mitochondrial function with a twofold increase in baseline oxygen consumption rate (2.25 Β± 0.03 vs. 1.13 Β± 0.1 nmol O2/min/106 cells) and FCCP-induced mitochondrial respiration (2.87 Β± 0.03 vs. 1.46 Β± 0.15 nmol O2/min/106 cells).

Conclusions: TGF-Ξ²1 induced differentiation of fibroblasts is accompanied by energetic remodeling of myofibroblasts with an increase in mitochondrial respiration and mitochondrial content.

β€’ Bioblast editor: Gnaiger E


  • ROUTINE respiration R'total (not baseline corrected for Rox): 1.13 Β± 0.1 nmol O2/min/106 cells is equivalent to 18.8 amolΒ·s-1Β·x-1
  • ET capacity E'total (not baseline corrected for Rox): 1.46 Β± 0.15 nmol O2/min/106 cells is equivalent to 24.3 amolΒ·s-1Β·x-1

Labels: MiParea: Respiration, mt-Biogenesis;mt-density 

Organism: Mouse  Tissue;cell: Fibroblast 

Coupling state: LEAK, ROUTINE, ET  Pathway: ROX 

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