Fig. 4. Compound 11 in Sacculatane diterpenoids from the Chinese liverwort Pellia epiphylla with protection against H O -induced apoptosis of PC12 cells

Fig. 4. Compound 11 protected PC12 cells against H2O2-induced cytotoxicity. (A) PC12 cells were treated with H2O2 (0–800 μM) for 2–6 h. The cell viability rate at an approximately percentage of 50% (200 μM, 4 h) was selected to be the experimental condition of model group. All data are expressed as the mean ± SD obtained of triplicate experiments. (B) Effects of 11 on cell viability (0–100 μM; 24 h) measured using the MTT assay. Compound 11 had no cytotoxicity up to 50 μM. (C) Effects of 11 (12.5–50 μM) on PC12 cells against H2O2-induced cell death. Cells were pretreated with indicated doses of 11 for 18 h and treated with 200 μM H2O2 for 4 h. CA (Catechin hydrate, 50 μM) was used as a positive control. (D) Effect of 11 on cell morphology detected by Olympus IX71 fluorescence imaging system. (E, F) Effect of 11 on intracellular ROS in PC12 cells induced by 200 μM H2O2 for 4 h. The fluorescence intensity of ROS in PC12 cells was examined using flow cytometry in the presence or absence of 11 (12.5–50 μM) or CA (50 μM). (G) Effect of 11 on the expressions of NQO1 and γ-GCS under stressed conditions with H2O2 (200 μM H2O2, 4 h). Cells were treated by indicated doses of 11 for 18 h, and then total cell lysates were analyzed by immunoblot analysis. (H) Effect of 11 on the relative expression level of NQO1 and γ-GCS under stressed conditions with H2O2 (200 μM H2O2, 4 h). All results were presented as the mean ± SD of three independent experiments. *P or #P <0.05, **P or ##P <0.01.