Curculigoside Protects against Excess-Iron-Induced Bone Loss by Attenuating Akt-FoxO1-Dependent Oxidative Damage to Mice and Osteoblastic MC3T3-E1 Cells

Figure 5

Effects of CUR on the expression of FoxO1 target gene as well as cell viability, apoptosis, and autophagy in MC3T3-E1 cells treated with excess iron. Osteoblastic MC3T3-E1 cells were treated with CUR, NAC, or FAC for 48 h, and after 2 h treatment, cotreated with FAC. (a) The expression of FoxO1 target genes p21, p27, cyclin D1, cyclin D2, Gadd45a, FasL, Bim, and Rab7 was detected with RT-qPCR analysis. (b) Cell viability was determined by MTT assay, and the morphological changes of cell confluence were observed under a phase-contrast microscope (×100). (c) The expression of autophagy proteins LC-3B, Beclin-1, and p62 was detected by Western blot and immunohistochemical staining (LC-3B). (d) Apoptosis-related proteins including caspase 3, Bcl-2, and Bax were detected by Western blot. (e) The mitochondrial membrane potential was detected with JC-1 assay kits. Results are represented as the (). , compared with the normal control group; ^#, ^## compared with the model group. N: normal control group, cells were treated with DMSO; M: model group, cells were treated with FAC (500 μM); DFO: cells were treated with DFO (100 μM) and FAC (500 μM); NAC: cells were treated with NAC (1000 μM) and FAC (500 μM); CUR: cells were treated with CUR (10 μM) and FAC (500 μM).

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