Figure 3.

Effects of HEP3 on proinflammatory cytokine productions in lipopolysaccharide (LPS)-activated RAW 264.7 cells, and effects on the d-galactose-induced HIEpiC senescent cells. Cell viability was measured by quantitative colorimetric methylthiazolyl tetrazolium (MTT) after incubation with HEP3 for 48 h [(A), a]; cells were preincubated with 0.05–0.20 mg/mL HEP3 for 4 h, and then treated with 1 μg/mL LPS for 24 h. Using an ELISA kit, tumor necrosis factor-α [(A), b], interleukin (IL)-1β [(A), c], IL-6 [(A), d], nitric oxide [(A), e], and inducible nitric oxide synthase [(A), f] in the supernatant were detected. [(B), a–e] The cells were treated with 40 mg/mL of d-galactose for 72 h combined with different concentrations of HEP3, and the number of senescent cells (blue-stained cells) was detected using β-galactosidase staining. [(B), f] The cells were treated with different concentrations of HEP for 24 h, and the cytotoxicity was detected by an MTT assay. [(B), g–i] The activities of malondialdehyde, total superoxide dismutase, and glutathione peroxidase of the cells. Values were means ± SDs of three independent experiments. ^#P < 0.05 vs the normal group, *P < 0.05, **P < 0.01 vs the model group, indicating significant differences compared with the model group.