目的 探讨天料木中异香豆素糖苷化合物4-hydroxy-2-{[（benzoyl） oxy]methyl}phenyl-β-D-glucopyranoside-6-benzoate （HPGB）对小鼠胚胎成骨细胞MC3T3-E1细胞活性的影响及其作用机制。方法 体外培养MC3T3-1细胞，用不同浓度的异香豆素糖苷化合物进行干预，采用MTT法测定MC3T3-1细胞增殖情况，碱性磷酸酶（ALP）测定细胞分化情况，悬液芯片技术检测、分析糖蛋白Dickkop （DKK-1）、骨保护素（OPG）、骨桥蛋白（OPN）、骨钙素（BGP）和核因子-κB受体活化因子配体（RANKL）蛋白表达水平。Real-time PCR法检测、分析DKK-1、RANKL mRNA的表达水平。结果 异香豆素糖苷化合物在3~300 μmol/L可促进MC3T3-E1细胞增殖和分化能力，具有浓度相关性；与正常对照组相比，异香豆素糖苷化合物可提高MC3T3-E1的ALP活性。蛋白结果表明，异香豆素糖苷化合物可显著上调MC3T3-E1细胞中OPG、OPN的表达，DKK-1、RANKL蛋白表达无明显变化，但显著提高了OPG/RANKL值。同时，mRNA结果显示异香豆素糖苷化合物可显著上调MC3T3-E1细胞中OPG mRNA表达，显著提高OPG/RANKL值。结论 异香豆素糖苷化合物可通过上调OPG、OPN的表达、提高OPG/RANKL值，促进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.

国家自然科学基金项目（81370096）；海南师范大学热带药用植物化学教育部重点实验室开放基金项目