Investigation of quercetin and hyperoside as senolytics in adult human endothelial cells

Abstract

Previously, quercetin has been reported to be a senolytic, a drug that selectively removes senescent cells, in HUVECs. However, we found neither quercetin nor Q3G was effective as a senolytic for adult human endothelial cells.

Fig 1

A) The experimental timeline. Cells were treated with either quercetin or Q3G for 48 hours before assays. B) A representative SABG staining comparing EP and SEN cells. Many of the SEN cells show characteristic flattened morphology, and are stained blue due to increased SABG activity. C) Representative image of the western blot and densitometry data for Lamin B1. *p<0.05 vs. baseline (N = 3 samples/group, per donor), T-test.

Fig 2

Relative cell abundance following quercetin A) and Q3G B) treatment of EP and SEN from three different donors. Cell proliferation was measured 48 hours after treatment, and was compared against the baseline count of cells frozen just prior to beginning treatment (t0). Values significantly lower than the t0 are indicated by asterisks. *p<0.05 vs. baseline (N = 16 wells/group, per donor), Kruskal-Wallis and Mann-Whitney U tests.

Fig 3

Live-Dead assay for quercetin (A) and Q3G (B) treatment of EP and SEN cells from the same three different donors. Data reflect the percentage of dead cells in response to different concentrations of quercetin or Q3G. N ≥ 6 wells/group, per donor. Approximately 1000 non-fragmented cells were scored for each N. *p<0.05 vs. baseline value. One-way ANOVA was used for Q3G data, which are normally distributed. Kruskal-Wallis and Mann-Whitney U tests were used for quercetin treatment to account for non-normality.

Fig 4. Manual quantification of SABG stain-positive cells.

SEN cells from the same three donors were treated with 6 μM quercetin and 100 μM Q3G followed by SABG staining. Manual counting was done to assess percent of SABG positive cells. *p<0.05 vs. baseline value (N = 12 wells/group, per donor, where each N ranged from 40 to 160 cells). T-test was used to determine statistical significance between control and test groups for each donor.