[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).

CONCLUSIONS

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.