张玉琴,孙承韬,王宏运,徐伟,褚克丹.天料木中异香豆素糖苷化合物对MC3T3-E1细胞增殖、分化及OPG/RANKL/RANK信号通路的影响[J].药物评价研究,2019,42(8):1515-1519
天料木中异香豆素糖苷化合物对MC3T3-E1细胞增殖、分化及OPG/RANKL/RANK信号通路的影响
Mechanism of isocoumarin glycoside from stems of Homalium paniculiflorum on activity of MC3T3-E1 cells via OPG/RANKL/RANK pathway
投稿时间:2019-01-27  
DOI:10.7501/j.issn.1674-6376.2019.08.005
中文关键词:  天料木;异香豆素糖苷化合物;MC3T3-1细胞;骨形成;OPG/RANKL/RANK信号通路
英文关键词:Homalium paniculiflorum How et Ko.;isocoumarin glycoside;MC3T3-E1 cells;bone formation;OPG/RANKL/RANK pathway
基金项目: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
作者单位E-mail
张玉琴 福建中医药大学药学院, 福建 福州 350122
海南师范大学热带药用植物化学教育部重点实验室, 海南 海口 571127 
 
孙承韬 福建中医药大学药学院, 福建 福州 350122  
王宏运 福建中医药大学药学院, 福建 福州 350122  
徐伟 福建中医药大学药学院, 福建 福州 350122 2000017@fjtcm.edu.cn 
褚克丹 福建中医药大学药学院, 福建 福州 350122  
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中文摘要:
      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。
英文摘要:
      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.
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