目的 探討天料木中異香豆素糖苷化合物4-hydroxy-2-{[（benzoyl） oxy]methyl}phenyl-β-D-glucopyranoside-6-benzoate （HPGB）對(duì)小鼠胚胎成骨細(xì)胞MC3T3-E1細(xì)胞活性的影響及其作用機(jī)制。方法 體外培養(yǎng)MC3T3-1細(xì)胞，用不同濃度的異香豆素糖苷化合物進(jìn)行干預(yù)，采用MTT法測(cè)定MC3T3-1細(xì)胞增殖情況，堿性磷酸酶（ALP）測(cè)定細(xì)胞分化情況，懸液芯片技術(shù)檢測(cè)、分析糖蛋白Dickkop （DKK-1）、骨保護(hù)素（OPG）、骨橋蛋白（OPN）、骨鈣素（BGP）和核因子-κB受體活化因子配體（RANKL）蛋白表達(dá)水平。Real-time PCR法檢測(cè)、分析DKK-1、RANKL mRNA的表達(dá)水平。結(jié)果 異香豆素糖苷化合物在3~300 μmol/L可促進(jìn)MC3T3-E1細(xì)胞增殖和分化能力，具有濃度相關(guān)性；與正常對(duì)照組相比，異香豆素糖苷化合物可提高M(jìn)C3T3-E1的ALP活性。蛋白結(jié)果表明，異香豆素糖苷化合物可顯著上調(diào)MC3T3-E1細(xì)胞中OPG、OPN的表達(dá)，DKK-1、RANKL蛋白表達(dá)無(wú)明顯變化，但顯著提高了OPG/RANKL值。同時(shí)，mRNA結(jié)果顯示異香豆素糖苷化合物可顯著上調(diào)MC3T3-E1細(xì)胞中OPG mRNA表達(dá)，顯著提高OPG/RANKL值。結(jié)論 異香豆素糖苷化合物可通過(guò)上調(diào)OPG、OPN的表達(dá)、提高OPG/RANKL值，促進(jìn)MC3T3-E1的增殖和分化。

Objective To explore the possible mechanism of isocoumarin glycoside from the stems of Homalium paniculiflorum on MC3T3-E1 cells activity.Methods The MC3T3-E1 cells were cultured and stimulated with various concentrations of isocoumarin glycoside for 48 h. And then, MTT and ALP assay were used to analyze the proliferation and differentiation of MC3T3-E1 cells, respectively. the protein expressions of Dickkop (DKK-1), Osteoprotegerin (OPG), Osteopontin (OPN), Bone gla protein (BGP) and Receptor activator of NK-κB ligand (RANKL) were detected by suspension chip technology. Furthermore,the mRNA expressions of OPG and RANKL were evaluated by Real-time PCR.Results Isocoumarin glycoside increased the proliferation and differentiation of MC3T3-E1 cells. Further mechanism analysis showed that isocoumarin glycoside significantly promoted the protein expression of OPG, OPN and OPG/RANKL ratio. While the protein expression of DKK-1 and RANKL had no obvious change. mRNA results showed that isocoumarin glycoside obviously promoted the mRNA expression of OPG and OPG/RANKL ratio. Conclusion Isocoumarin glycoside increased MC3T3-E1 cells proliferation and differentiation through OPG/RANKL/RANK pathway, and and induce bone formation.

國(guó)家自然科學(xué)基金項(xiàng)目（81370096）；海南師范大學(xué)熱帶藥用植物化學(xué)教育部重點(diǎn)實(shí)驗(yàn)室開(kāi)放基金項(xiàng)目