Abstract
BackgroundPuerarin (daidzein 8-C-glucoside) has potential on preventing osteoporosis. This study aims to investigate the effects of puerarin on osteogenesis and adipogenesis in vitro.MethodsCCK-8 assay, alkaline phosphatase (ALP) activity and Alizarin Red S were used to measure the effects of puerarin on proliferation, osteoblastic differentiation, and mineralization in osteoblast-like MC3T3-E1 cells. The effects of puerarin on adipogenesis were measured by Oil Red O staining and intracellular triglyceride level in preadipocyte 3T3-L1 cells. The mRNA and protein levels of osteogenesis- and adiopogenesis-related factors were detected by qRT-PCR and western blot, respectively. Further, the secreted osteocalcin levels and nuclear translocation of β-catenin were detected by ELISA and immunofluorescence assay, respectively.ResultsAs to osteogenesis, puerarin could stimulate proliferation (1 μM, P = 0.012; 10 μM, P = 0.015; 20 μM, P = 0.050), ALP activity (20 μM, P = 0.008) and calcium nodule formation (20 μM, P = 0.011) in a dose-dependent manner. Puerarin (20 μM) promoted osteocalcin secretion (P = 0.004) and the protein expression of both osteopontin (P = 0.001) and osteoprotegerin (P = 0.003). As to adipogenesis, puerarin suppressed adipocytes formation and intracellular triglyceride level (P = 0.001). In addition, puerarin (20 μM) decreased the mRNA and protein levels of CCAAT/enhancer binding protein α (P = 0.001, P = 0.002), proliferator-activated receptor γ (P = 0.005, P = 0.003), and adipocyte lipid-binding protein 4 (P = 0.001, P = 0.001). Moreover, phosphorylation of AKT1-Ser437 (10 μM, P = 0.003; 20 μM, P = 0.007) and GSK-Ser9 (10 μM, P = 0.005; 20 μM, P = 0.003), and the nuclear translocation of β-catenin (10 μM, P = 0.006; 10 μM, P = 0.002) were increased in 3T3-L1 cells treated by puerarin.ConclusionPuerarin promoted osteogenesis and inhibited adipogenesis in vivo, and Akt/GSK-3β/β-catenin signaling pathway was involved in the suppression of adipogenesis.
Highlights
IntroductionPuerarin (daidzein 8-C-glucoside) has potential on preventing osteoporosis
MC3T3-E1 cells MC3T3-E1 osteoblastic cells were cultured in puerarin at various concentrations (0, 0.1, 1, 10 and 20 μM) for 48 h
Puerarin further increased the OC levels expression to 36.4% at 10 μM dosage (P = 0.003), and 42.2% at 20 μM dosage (P = 0.004), compared with ctl+ group (Figure 2E). These results suggested that puerarin could promote proliferation, alkaline phosphatase (ALP) activity, mineralization and OC protein secretion in MC3T3-E1 cells
Summary
Puerarin (daidzein 8-C-glucoside) has potential on preventing osteoporosis. This study aims to investigate the effects of puerarin on osteogenesis and adipogenesis in vitro. The first-line therapeutic strategy for postmenopausal osteoporosis is estrogen replacement therapy (ERT) to prevent bone loss and increase bone formation [3], through the enhancement of osteoblast differentiation and bone formation [4,5,6], and inhibition of osteoclast. Some reports demonstrated that daidzein promoted osteogenic, inhibited adipogenic differentiation and exhibited preventive activity on bone loss in OVX animals [24,25]. This study aims to investigate the dual effects and molecular mechanism of puerarin on osteogenesis and adipogenesis in vitro
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