摘要
目的研究化合物丹皮酚对大鼠骨髓间充质干细胞(BMMSCs)增殖及成骨向分化的作用。方法体外培养大鼠BMMSCs,使用不同浓度(0、0.5、5、15、30、60、120μmol/L)丹皮酚作用于细胞,CCK-8法检测细胞增殖活性;利用碱性磷酸酶染色和茜素红染色检测BMMSCs成骨向分化效率;RT-PCR检测成骨向分化相关基因(Alp、Runx2、Ocn、Opn)mRNA水平;Western blot检测RUNX2、ALP蛋白表达量。结果丹皮酚在第3和第5天时可明显促进细胞生长,且在15μmol/L和30μmol/L浓度下作用最明显;碱性磷酸酶和茜素红染色结果显示丹皮酚可促进BMMSCs成骨向分化,其中30μmol/L时细胞矿化最显著;丹皮酚增加成骨相关基因(Runx2、Alp、Ocn、Opn)mRNA水平并促进RUNX2、ALP蛋白表达。结论丹皮酚可促进BMMSCs的增殖和成骨向分化,有望为牙周炎骨缺损的治疗提供一个新的候选化合物。
Objective To examine the effect of paeonol on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells(BMMSCs).Methods BMMSCs of rats were cultured in vitro,and the cells were treated with different concentrations of paeonol.CCK-8 method was used to detect cell proliferation activity.Osteogenic differentiation efficiency of BMMSCs cells was examined by alkaline phosphatase staining and alizarin red staining.mRNA levels of osteogenic differentiation-related gen es(Alp,Runx2,Ocn,Opn)were determined by real-time fluorescent quantitative polymerase chain reaction.ALP and RUNX2 protein expression was analyzed By Western blot.Results Paeonol significantly promoted cell growth on days 3 and 5,and the effect was m ost pronounced at 15μmol/L and 30μmol/L.Alkaline phosphatase and alizarin red staining showed that paeonol promoted osteogenic differentiation of BMMSCs,with the most prominent cell mineralization at 30μmol/L.Paeonol increased mRNA levels of osteogenic genes(Runx2,Alp,Ocn,Opn)and promoted RUNX2 and ALP protein expression.Conclusion Paeonol can promote the proliferation and osteogenic differentiation of BMMSCs,and it is expected to provide a new candidate compound for the treatment of periodontitis bone defects.
作者
王娟
祁胜财
周蓉
刘海霞
蔡怡闻
徐远志
陈万涛
WANG Juan;QI Sheng-cai;ZHOU Rong;LIU Hai-xia;CAI Yi-wen;XU Yuan-zhi;CHEN Wan-tao(Wei Fang Medical University School of Stomatology,Weifang 261000,China)
出处
《北京口腔医学》
CAS
2020年第1期6-11,共6页
Beijing Journal of Stomatology
基金
上海动物模型专项研究(16140903300)。
关键词
丹皮酚
骨髓间充质干细胞
细胞增殖
成骨向分化
Paeonol
Bone marrow mesenchymal stem cells
Cell proliferation
Osteogenic differentiation
