Fig. 6

Mitochondrial metabolic activity is reduced and impairs mitochondrial function after treatment with plasma. (A) There was noted an energy deficiency accessed by MTT assay both in PO and PE compared to the control group after 10 days of treatment. (B) After 10 days of treatments, flow cytometry analysis revealed (left) decreased membrane potential (MTR), but (right) augmented mitochondrial biomass (MTG) was detected in PO compared to PE and control groups. (C) Decreased mitochondrial function assessed by the MTR/MTG ratio after exposure to PO when compared to PE and control treatments after 10 days. (D) Representative images (scale bar 200 μm) acquired by fluorescence microscopy with MTR and MTG staining after 10 days of treatment. (E) (Left) High-resolution respirometry was performed and (right) decreased oxygen consumption was observed in ADSC treated with PO compared to the PE group. (F) There was no difference in O2^•− expression after 10 days of treatments. (G) Relative mtDNA copies were increased in PO after 10 days of treatment. Data presented as mean and standard deviation (SD). Dashed lines represent positive control of senescence assessed by 300 μM of hydrogen peroxide (H₂O₂) treated for 3 h two days before experiments. Differences were considered when p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), or p < 0.0001 (****), evaluated by one-way ANOVA test followed by Tukey post-test, with a confidence interval of 95%. Abbreviations; MFI: median fluorescence intensity; ; mtDNA: mitochondrial DNA; MTG: MitoTracker™ Green FM; MTR: MitoTracker™ Red CMXRos; MTT: [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]; nucDNA: nuclear DNA; ; PE: pool plasma of eutrophic individuals supplemented in the complete culture medium; PO: pool plasma of individuals with obesity supplemented in the complete culture medium; O2^•−: Mitochondrial superoxide