[Involvement of Wnt/β-catenin signaling in the osteogenesis of bone marrow mesenchymal stem cells induced by drynaria total flavonoids].

OBJECTIVE: To explore the effects of the expression of Wnt/β-catenin signaling factor mRNA during drynaria total flavonoids on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).

METHODS: The BMSCs were isolated from SD rats by whole bone marrow culture method and purified by passage. And the P3 BMSCs were intervened with 100 µg/ml drynaria total flavonoids. At Day 21, mineralized staining was performed. At Days 7, 14, 21 and 28 post-intervention, the activity of alkaline phosphatase (ALP) was detected and polymerase chain reaction (PCR) used to detect the expressions of Wnt/β-catenin signaling pathway related factors β-catenin, LEF-1 and cycline D mRNA.

RESULTS: At each time point post-intervention, comparing the ALP activity in cell supernatant between two group, the drynaria total flavonoids group was higher than the blank control group (7 d: 11.10 ± 0.08 vs 1.61 ± 0.14; 14 d: 24.62 ± 0.34 vs 1.64 ± 0.04; 21 d: 18.41 ± 0.06 vs 1.53 ± 0.04; 28 d: 14.9 ± 0.14 vs 1.52 ± 0.04; all P < 0.01). At Day 21, upon staining with alizarin red, the drynaria total flavonoids group was positive while the blank control group negative. At Day 14, the expression of β-catenin mRNA was higher in the drynaria total flavonoids group higher than that in the blank control group (0.357 ± 0.063 vs 0.174 ± 0.013, P < 0.05). At Day 7, the expressions of LEF-1 and cycline D mRNA were higher in the drynaria total flavonoids group than those in the blank control group (LEF-1 0.0611 ± 0.0002 vs 0.0345 ± 0.0131; cycline D 0.1510 ± 0.0255 vs 0.0718 ± 0.0294, all P < 0.05).

CONCLUSION: Drynaria total flavonoids induce BMSCs to differentiate into osteoblasts. And it is accompanied with the altered expression of Wnt/β-catenin signaling pathway related factor mRNA.