[關(guān)鍵詞]
[摘要]
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)對小鼠胚胎成骨細胞MC3T3-E1細胞活性的影響及其作用機制。方法 體外培養(yǎng)MC3T3-1細胞,用不同濃度的異香豆素糖苷化合物進行干預(yù),采用MTT法測定MC3T3-1細胞增殖情況,堿性磷酸酶(ALP)測定細胞分化情況,懸液芯片技術(shù)檢測、分析糖蛋白Dickkop (DKK-1)、骨保護素(OPG)、骨橋蛋白(OPN)、骨鈣素(BGP)和核因子-κB受體活化因子配體(RANKL)蛋白表達水平。Real-time PCR法檢測、分析DKK-1、RANKL mRNA的表達水平。結(jié)果 異香豆素糖苷化合物在3~300 μmol/L可促進MC3T3-E1細胞增殖和分化能力,具有濃度相關(guān)性;與正常對照組相比,異香豆素糖苷化合物可提高MC3T3-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細胞中OPG、OPN的表達,DKK-1、RANKL蛋白表達無明顯變化,但顯著提高了OPG/RANKL值。同時,mRNA結(jié)果顯示異香豆素糖苷化合物可顯著上調(diào)MC3T3-E1細胞中OPG mRNA表達,顯著提高OPG/RANKL值。結(jié)論 異香豆素糖苷化合物可通過上調(diào)OPG、OPN的表達、提高OPG/RANKL值,促進MC3T3-E1的增殖和分化。
[Key word]
[Abstract]
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.
[中圖分類號]
[基金項目]
國家自然科學(xué)基金項目(81370096);海南師范大學(xué)熱帶藥用植物化學(xué)教育部重點實驗室開放基金項目