Effects of Xinshang Xuduan Decoction on Osteoblast Proliferation and Bone Morphogenetic Protein in Vitro / 广州中医药大学学报

ABSTRACT

Objective To investigate the effects of Xinshang Xuduan Decoction(XXD, mainly composed of Rhizoma Drynariae, Radix Dipsaci, and Eupolyphaga seu Steleophaga) on the proliferation and differentiation of rat osteoblasts and bone morphogenetic protein 2(BMP-2)expression in the osteoblasts . Methods The animal serum containing XXD was prepared by serum pharmacological method and then was mixed together with α-MEM for the cell culture. Osteoblasts were isolated from the skull bone of SD neonatal rats by collagenase digestion and were identified by alkaline phosphatase(ALP) staining and alizarin red staining. The third and fourth generations of osteoblasts were treated with XXD at the volume fraction of 5%, 10%, 20% for 24, 48 , 72 hours respectively, and then the proliferation of the cells was evaluated by Cell Counting Kit-8(CCK-8). In the other test, osteoblasts were cultured with blank control serum, and 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae, respectively, and then the ALP activity was examined by using ALP assay kit and the expression of BMP-2 was investigated by enzyme-linked immunosorbent assay(ELISA). Results XXD had a dose- and time-dependent effect on the proliferation of rat osteoblasts in a suitable volume fraction range from 5% to 20%, and the effect of XXD at 10% was the best. Compared with the blank control serum group, ALP activity was increased in the cells treated with 10% serum containing XXD, Radix Dipsaci, and Rhizoma Drynariae(P<0.01). On culturing day 7, the expression of BMP-2 was increased in 10% XXD group and Rhizoma Drynariae group(P<0.05). Conclusion XXD can increase the ALP activity and BMP-2 expression in the osteoblasts in vitro, so does the single herb of Rhizoma Drynariae. And their therapeutic mechanism in promoting the healing of fractures may be related with the enhancement of osteogenesis of osteoblasts.