Objective: To investigate the effect of icariin on the proliferation, differentiation, and the mRNA expressions of Cbfalpha1, BMP2, BMP4 of rat osteoblasts.
Methods: Primary rat osteoblastic cells were obtained by sequentia collagenase/trypsin enzyme digestion from calvarial bones of new born (within 24 h) SD rats and were identified by Alkaline phosphatase and alizarin red staining. The passage 3-5 cells were treated with icariin at the concentration of 0 mol/L, 10(-8) mol/L, 10(-7) mol/L, 10(-6) mol/L, 10(-5) mol/L, 10(-4) mol/L for 24 h, 48 h, 72 h, and the proliferation of the cells was measured by CCK-8 assay. The proliferation index was detected by Flow Cytometry and the activity of alkaline phosphatase was determined by p-Nitrophenyl phosphate (pNPP) method after being treated with icariin at the concentration mentioned above for 48 h. The total cellular RNA was extracted 48 h after being treated with icariin at the concentration of 10(-6) mol/L, and the expressions of Cbfalpha1, BMP2, BMP4 mRNA were examined by real-time PCR.
Results: Icariin showed no effect on the proliferation of osteoblasts, but improved ALP activity. The Cbfalpha1, BMP2, BMP4 mRNA were significantly upregulated after icariin treatment.
Conclusion: Icariin could promote the differentiation ability of rat osteoblasts through upregulating the Cbfalpha1, BMP2, BMP4 mRNA expressions.