Objective The extract of Polygonum Multiflorum is to be derived by extracting, separating and purifying under the guidance of bioactivity.To observe the influence of Polygonum Multiflorum s extractive on the proliferation and osteogenic differentiation of bone marrow stromal cell (BMSCs) and on the skull bone mass of a bone-loss model induced by glucocorticoid in Zebrafish.Methods The Polygonum Multiflorum was extracted, separated , purified and assayed in application of column chromatography and modern spectroscopic analytical techniques.And the effect of proliferation, alkaline phosphatase activity and osteocalcin levels of BMSCs was aspectively tested by the method of MTT, PNPP and the two-step double antibody sandwich enzyme-linked immunosorbent assay.So that the extract s effect of the proliferation and osteogenic differentiation of BMSCs and it s relationship of time-effect and dose-effect were observed.A bone-loss model was established through the bone-loss induced by the dexamethasone glucocorticoid in Zebrafish.The drug, which was preliminary evaluated on the vivo efficacy through quantitative analysis on the area and it s cumulative optical density.Results There were 21 compounds separated and purified-from 50% ethanol elution fraction of Polygonum Multiflorum, of which the chemical structure of seven ones were identified.Compound 21 : emodin-1-O-β-D-glucopyranoside; Compound 5,6: Polygonum 1; Compound 1: Polygonum 2; Compound 8: 2,3,5,4-tetrahydroxy-stilbene-2-O-β-D-glucoside; Compound 18: 2,3,5,4-tetrahydroxy-stilbene-2-O-β-D-glucoside; Compound 10: (-)-Catechin.Compound 1, 3, 8, 10, and 18 promote proliferation and osteogenic differentiation of BMSCs.Compound 2, 8, 10, and 18 increased osteocalcin secretion.Compound 2, 3and 8 had prevention effect on 10μmol/L dexamethasone-induced zebrafish bone loss.Compound 2, 5 and 18 decrease the expression of WIF 1 and GSK3 β.Conclusions Seven compounds were obtained from Polygonum Multiflorum s extractive, and two compounds containing stilbene glucoside fragment were new compounds.50% ethanol-eluted fraction of Polygonum Multiflorum contains several active ingredients of anti-osteoporosis.From these, compounds 1, 2, 3, 8 and 18 could promote BMSCs differentiation into osteoblasts.Compounds 2, 3 and 8 could prevent bone loss induced by 10 μ mol/L of dexamethasone in zebrafish model, the effects may be associated with the regulation of Wnt/β-catenin signaling pathway.