FIGURE 5.

Metformin ameliorated LPS-triggered hyper-inflammation in DOX-induced senescent HUVECs. (A) Schematic diagram of the experimental design. HUVECs were treated for 24 h with 0.5 µM DOX ±5 mM metformin (added 24 h before DOX) or left untreated. Thereafter, DOX was removed and the cells were incubated in DOX-free media with or without metformin for an additional 72 h. The media were changed so that the assessed SASP factors in the media reflect only the effect of LPS. Then, cells were stimulated with LPS (30 ng/mL) for an additional 24 h. Thereafter, conditioned media were collected and the protein expression of SASP factors including (B) IL-6, (C) CXCL2, (D) MMP-3, (E) TNF-α, and (F) MCP-3 was determined by Luminex (n = 4). Values were normalized to the protein concentration of the cells determined by BCA. Expressed values are presented as mean ± SEM. Data were analyzed by two-way ANOVA followed by a Tukey multiple comparisons test. * compared to different treatment within the same group; * p <0.05, ** p <0.01, **** p < 0.0001. # compared to non-senescent cells with the same treatment; # p <0.05, #### p < 0.0001.