Figure 3.

Icariin alleviates TM4 cells injury induced by D-galactose (D-gal). (A) Cellular viability was detected using MTT assay. TM4 cells at a concentration of 3 × 10⁴ cells/well in 96-well plates were pretreated with icariin (0.01–8 μM) or DMSO (0.1%) for 20 h and then incubated with 100 mM D-gal for 60 h. (B–D) TM4 cells at 5 × 10⁵/well in 6-well plates were pretreated with icariin (0.5–1 μM) for 20 h and then incubated with 100 mM of D-gal for 60 h. (B) Representative images of SA-β-gal staining of cells (scale bar = 50 μm). (C) The percentages of SA-β-gal-positive cells from a total of 500 cells. (D) The relative protein expression levels of GDNF, PLZF, BMP4, and SCF were measured using western blotting analysis. All values are expressed as means ± SEM of three independent experiments. ^##P < 0.01 versus control; ^*P < 0.05, ^**P < 0.01 versus D-gal-stimulated group.