Fig. 4.

Metformin has beneficial effects on function restoration in senescent VSMCs. The migration of Ang II-induced senescent VSMCs was determined by the wound-healing assay. C, Semi-quantitative analysis of wound area healing at a fixed location was performed with Image J. VSMCs underwent incubation with or without Ang II (2 μM) for 24 h with metformin (200 μM) treatment or not (one-way ANOVA, Tukey post hoc; n = 4). B, D, E, and F, The expression levels of PCNA, SM-22α, and calponin in Ang II-induced senescent VSMCs were assessed by immunoblot, using α-tubulin for normalization (one-way ANOVA, Tukey post hoc; n = 3). G-H, Flow cytometry showed G0/G1 cell-cycle arrest was increased significantly in Ang II-treated cells, and this effect was alleviated by metformin treatment, while metformin increased S phase cells that were inhibited by Ang II treatment (one-way ANOVA, Tukey post hoc; n = 3). Data are mean ± SD. *P < 0.05, ^**P < 0.01, and ^***P < 0.001 versus control group or for indicated comparisons.