二氢杨梅素对SHED活性及其成骨分化能力的影响
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摘要
目的探讨二氢杨梅素(DMY)对人乳牙牙髓干细胞(SHED)活性及成骨分化能力的影响。方法体外分离培养鉴定SHED,分别检测不同浓度的DMY对SHED的细胞活性、碱性磷酸酶(ALP)活性以及成骨相关基因Runt相关转录因子2(Runx2)和骨钙素(OCN) mRNA的表达的影响。结果 CCK-8检测结果显示,与对照组比较,0. 02、0. 1、0. 5、2、10及50μmol/L浓度的DMY对SHED的增殖能力差异无统计学意义;10、50μmol/L浓度的DMY ALP活性较对照组高(P <0. 05),0. 1、0. 5、2、10及50μmol/L DMY组的Runx2及0. 5、2、10、50μmol/L DMY组OCN成骨诱导前后相对倍增数均高于对照组(P <0. 05),并呈现一定的浓度依赖性。结论 0. 02、0. 1、0. 5、2、10及50μmol/L的(DMY)对SHED活性无明显影响,一定浓度的DMY可促进SHED成骨分化的能力。
Objective To observe the effect of Dihydromyricetin( DMY) on activity and osteogenic differentiation of stem cells from human exfoliated deciduous teeth( SHED) in vitro. Methods SHED were gained by Enzymatic digestion in vitro. The cellular viability,alkaline phosphotase activity,osteogenic differentiation related genes,such as Runt-related transcription factor 2( Runx2) and osteocalcin( OCN) were observed after different concentration of DMY act upon SHED. Results 0. 02,0. 1,0. 5,2,10 and 50 μmol/L concentration of DMY had no cytotoxicity to SHED. 10,50 μmol/L concentration of DMY increased the activity of alkaline phosphatase( P < 0. 05). 0. 1,0. 5,2,10 and 50 μmol/L concentration of DMY increased the mRNA expressions of Runx2,0. 5,2,10 and 50 μmol/L concentration of DMY increased the mRNA expressions of OCN which were in a concentration-dependent manner( P< 0. 05). Conclusion 0. 02,0. 1,0. 5,2,10,50 μmol/L concentration have no significant effect on the activity of SHED. A certain concentration of DMY can enhance the osteogenic differentiation of SHED.
Objective To observe the effect of Dihydromyricetin( DMY) on activity and osteogenic differentiation of stem cells from human exfoliated deciduous teeth( SHED) in vitro. Methods SHED were gained by Enzymatic digestion in vitro. The cellular viability,alkaline phosphotase activity,osteogenic differentiation related genes,such as Runt-related transcription factor 2( Runx2) and osteocalcin( OCN) were observed after different concentration of DMY act upon SHED. Results 0. 02,0. 1,0. 5,2,10 and 50 μmol/L concentration of DMY had no cytotoxicity to SHED. 10,50 μmol/L concentration of DMY increased the activity of alkaline phosphatase( P < 0. 05). 0. 1,0. 5,2,10 and 50 μmol/L concentration of DMY increased the mRNA expressions of Runx2,0. 5,2,10 and 50 μmol/L concentration of DMY increased the mRNA expressions of OCN which were in a concentration-dependent manner( P< 0. 05). Conclusion 0. 02,0. 1,0. 5,2,10,50 μmol/L concentration have no significant effect on the activity of SHED. A certain concentration of DMY can enhance the osteogenic differentiation of SHED.
引文
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[6]郭航,王永军,谢鹏,等.日粮添加二氢杨梅素对肉仔鸡肠黏膜形态结构、碱性磷酸酶及生产性能的影响[J].中国饲料,2008,6:19-22.
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[8] Qi S,Xin Y,Guo Y,et al. Ampelopsin reduces endotoxic inflammation via repressing ros-mediated activation of PI3K/AKT/NFkappa B signaling pathways[J]. Int Immunopharmacol,2012,12(1):278-87.
[9] Zhou Y,Shu F,Liang X,et al. Ampelopsin induces cell growth inhibition and apoptosis in breast cancer cells through ros generation and endoplasmic reticulum stress pathway[J]. PLo S One,2014,9(2):e89021.
[10]侯小龙,王文清,施春阳,等.二氢杨梅素药理作用研究进展[J].中草药,2015,46(4):603-9.
[11] Zheng Y,Liu Y,Zhang C M,et al. Stem cells from deciduous tooth repair mandibular defect in swine[J]. J Dent Res,2009,88(3):249-54.
[12] Fu X,Jin L,Ma P,et al. Allogeneic stem cells from deciduous teeth in treatment for periodontitis in miniature swine[J]. J Periodontol,2014,85(6):845-51.
[13]周防震,郑小江,张晓元.藤茶活性成分二氢杨梅素对人胚肾细胞和肝细胞增殖的影响[J].中成药,2015,37(12):2756-8.
[14]郑文龙,吴涛,邹多宏,等.犬牙槽骨干细胞与髂骨骨髓干细胞体外成骨能力的比较[J].安徽医科大学学报,2016,51(9):1252-7.
[15] Gao R T,Zhan L P,Meng C,et al. Homeobox B7 promotes the osteogenic differentiation potential of mesenchymal stem cells by activating RUNX2 and transcript of BSP[J]. Int J Clin Exp Med,2015,8(7):10459-70.
[2] Akiyama K,Chen C,Gronthos S,et al. Lineage differentiation of mesenchymal stem cells from dental pulp,apical papilla,and periodontal ligament[J]. Methods Mol Biol,2012,887:111-21.
[3]赵晓丽,陈金晶,司书毅,等.野黄芩苷体外促成骨分化作用研究[J].中国医药生物技术,2016,11(1):27-31.
[4] Yin L H,Cheng W X,Qin Z S,et al. Effects of ginsenoside Rg-1 on the proliferation and osteogenic differentiation of human periodontal ligament stem cells[J]. Chin J Integr Med,2015,21(9):676-81.
[5]陈夏静,姚汉玲,张秋琼,等.大叶蛇葡萄中蛇葡萄素含量的高效液相色谱法测定[J].时珍国医国药,2010,21(5):1053-4.
[6]郭航,王永军,谢鹏,等.日粮添加二氢杨梅素对肉仔鸡肠黏膜形态结构、碱性磷酸酶及生产性能的影响[J].中国饲料,2008,6:19-22.
[7] Zhang W,Wang S,Yin H,et al. Dihydromyricetin enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells in vitro partially via the activation of Wnt/β-catenin signaling pathway[J]. Fund Clin Pharmacol,2016,30(6):596-606.
[8] Qi S,Xin Y,Guo Y,et al. Ampelopsin reduces endotoxic inflammation via repressing ros-mediated activation of PI3K/AKT/NFkappa B signaling pathways[J]. Int Immunopharmacol,2012,12(1):278-87.
[9] Zhou Y,Shu F,Liang X,et al. Ampelopsin induces cell growth inhibition and apoptosis in breast cancer cells through ros generation and endoplasmic reticulum stress pathway[J]. PLo S One,2014,9(2):e89021.
[10]侯小龙,王文清,施春阳,等.二氢杨梅素药理作用研究进展[J].中草药,2015,46(4):603-9.
[11] Zheng Y,Liu Y,Zhang C M,et al. Stem cells from deciduous tooth repair mandibular defect in swine[J]. J Dent Res,2009,88(3):249-54.
[12] Fu X,Jin L,Ma P,et al. Allogeneic stem cells from deciduous teeth in treatment for periodontitis in miniature swine[J]. J Periodontol,2014,85(6):845-51.
[13]周防震,郑小江,张晓元.藤茶活性成分二氢杨梅素对人胚肾细胞和肝细胞增殖的影响[J].中成药,2015,37(12):2756-8.
[14]郑文龙,吴涛,邹多宏,等.犬牙槽骨干细胞与髂骨骨髓干细胞体外成骨能力的比较[J].安徽医科大学学报,2016,51(9):1252-7.
[15] Gao R T,Zhan L P,Meng C,et al. Homeobox B7 promotes the osteogenic differentiation potential of mesenchymal stem cells by activating RUNX2 and transcript of BSP[J]. Int J Clin Exp Med,2015,8(7):10459-70.