Formononetin inhibits p53 signaling pathway activation to delay cellular senescence and ameliorates diabetic kidney disease

Figure 4 Formononetin intervention delays senescence of MPC-5 cells via p53 signaling pathway. A: Western blot bands of p53 signaling pathway; B: Dual-luciferase reporter gene assay demonstrating that formononetin (FN) intervention significantly reduces p53 transcriptional activity; C-F: Western blot analysis reveals that FN intervention significantly increases MDM2 and CCND1 protein levels and decreases p53 and p21 protein levels; G: Β-galactosidase (β-GAL) indicator cell staining photography (100 ×); H: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay showing that MDM2 silencing weakens the protective effects of FN on high glucose-induced MPC-5 cell viability; I and J: Cell staining and cell supernatant β-GAL activity assay showing that MDM2 silencing eliminates the reduction in β-GAL activity induced by FN. n = 3 per group. ^aP < 0.05 vs control groups; ^bP < 0.01 vs control groups; ^cP < 0.05 vs high glucose groups; ^dP < 0.01 vs high glucose groups. NS: Not significant; FN: Formononetin; HG: High glucose; H-FN: High dose of formononetin; C: Control; shNC: Short hairpin negative control; shMDM2: Short hairpin RNA targeting MDM2.