Fig. 5
Chronic exposure to plasma triggers inflammation and is associated with p38MAPK/NF-κB axis in senescent ADSC. Flow cytometry analysis of (A) NF-κB p65 subunit phosphorylation levels (left, MFI, and right, %) were higher in PO when compared to control after 10 days of culture. Quantification of (B) IL-6 secretion and C) IL-8 secretion were higher in PO and PE groups in comparison to control treatment after 10 and 18 days. (D) γ-H2AX protein expression after 2 days (left, MFI, and right, %) of ADSC was similar among treatments with plasma or control. (E) p38MAPK phosphorylation levels were augmented (left, MFI, and right, %) in PO compared to both PE and control groups after 10 days of treatments. Dashed lines represent ADSC incubated with 300 μM of hydrogen peroxide (H2O2) for 3 h two days before experiments, treated as a positive control of senescent cells. Data presented as mean and standard deviation (SD). 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: Phospho-p38MAPK: phosphorylated p38 mitogen-activated protein kinase; Phospho-p65: phosphorylated nuclear factor NF-κB p65 subunit; PE: pool of plasma from eutrophic individuals supplemented in culture medium; PO: pool of plasma from individuals with obesity supplemented in culture medium; SASP: senescence-associated secretory phenotype