摘要
目的探讨大豆黄素促进脱落乳牙干细胞(SHED)成骨的作用及其机制。方法对体外培养的第3代SHED分别用100、10和1μmol.L-1的大豆黄素培养基进行培养,以普通培养基作为对照,于3、6、9 d检测其碱性磷酸酶(AKP)活性,于7、14、21 d检测其骨钙蛋白(OC)的质量,以RT-PCR检测其3、6、9 d的核心结合因子(cbf)-α1 mRNA的表达。结果 SHED经大豆黄素干预培养3 d,10μmol.L-1的大豆黄素显著提高了细胞内AKP的质量(与对照组比较,P<0.05);培养6～9 d,3个浓度的大豆黄素均显著提高了AKP的表达量(与对照组比较,P<0.001)。中高浓度(10和100μmol.L-1)的大豆黄素可在成骨分化的中期(14 d)显著提高SHED内OC的质量(与对照组比较,P<0.001),21 d时各浓度的大豆黄素均可促进OC的表达(与对照组比较,P<0.001)。SHED培养3 d,10和100μmol.L-1大豆黄素明显上调其cbf-α1 mRNA的表达,1μmol.L-1的大豆黄素部分上调了cbf-α1mRNA的表达;在第6和9天,1、10和100μmol.L-1的大豆黄素均促进了cbf-α1 mRNA的表达,对照组在各时间点均呈阴性。结论大豆黄素可促进SHED向成骨方向分化,其机制可能与上调cbf-α1基因表达有关。
Objective To investigate the effects and mechanism of daidzein on osteodifferentiation of stem cells derived from human exfoliated deciduous teeth（SHED）.Methods The 3rd passage SHED were incubated with 1, 10,100 μmol·L-1 daidzein and normal culture medium（control）.Then alkaline phosphatase（AKP） activities were examined at 3,6 and 9 d,and osteocalcin（OC） in each group was detected at 7,14 and 21 d.In addition,core-binding factor（cbf）-α1 mRNA expression were detected by RT-PCR at 3,6 and 9 d after culture.Results 3 d later,AKP activities in SHED were enhanced by 10 μmol·L-1 daidzein（vs.control,P0.05）,all of three concentrations（1,10,100 μmol·L-1） daidzein improved AKP activities significantly at 6 and 9 d（vs.control,P0.001）. Medium and high dose（10,100 μmol·L-1） daidzein could enhance OC expression in SHED at 14 d after induced culture（vs.control,P0.001）,and all of the doses daidzein increased OC expression at 21 d（vs.control,P0.001）.Additionally,RT-PCR showed that 10 and 100 μmol·L-1 daidzein could significantly increase cbf-α1 mRNA expression at the 3rd,while 1 μmol·L-1 daidzein only slightly increase cbf-α1 mRNA expression;at the 6th and 9th d,however,three concentration daidzein increased cbf-α1 mRNA expression obviously.Conclusion Daidzein could stimulate osteodifferentiation of SHED in vitro,and the possible mechanism is its role of promoting the expression of cbf-α1 gene.
出处
《国际口腔医学杂志》
CAS
2012年第3期308-311,316,共5页
International Journal of Stomatology
关键词
大豆黄素
人乳牙牙髓干细胞
成骨细胞分化
机制
daidzein
stem cell derived from human exfoliated deciduous teeth
osteoblast differentiation
mechanism
