山西医科大学学报2023，Vol.54Issue(12)：1638-1646,9.DOI:10.13753/j.issn.1007-6611.2023.12.013
7,8-二羟基黄酮通过Wnt/β-catenin信号通路调控人牙周膜干细胞成骨分化
7,8-dihydroxyflavone regulates osteogenic differentiation of human periodontal ligament stem cells through Wnt/β-catenin signaling pathway
摘要
Abstract
Objective To investigate the effect of 7,8-dihydroxyflavone(7,8-DHF)on the osteogenic differentiation of human perio-dontal ligament stem cells(hPDLSCs)and its mechanism.Methods The hPDLSCs isolated from the periodontal tissue of healthy human were treated with different concentrations of 7,8-dihydroxyflavone(0,0.5,1,5,10,20,40 μmol/L),and then the activity of hPDLSCs was detected by MTT assay.The hPDLSCs were divided into 5 groups;control group,low-dose group,medium-dose group,high-dose group,and XAV939(a Wnt/β-catenin signaling pathway inhibitor)group.The hPDLSCs in control group were only cultured with osteogenic induction solution.The hPDLSCs in low-dose group,medium-dose group and high-dose group were cultured with osteogenic induction solutions containing 0.5,1,5 µmol/L 7,8-dihydroxyflavone,respectively.The hPDLSCs in XAV939 group were cultured with osteogenic induction solution containing 5 μmol/L 7,8-dihydroxyflavone and 10 μmol/L XAV939 at the same time.The hPDLSCs were cultured for 21 d in each group,and the medium was changed every two days.Alkaline phosphatase(ALP)activity was detected and calcified nodule formation was detected by Alizarin red staining.The mRNA expression levels of Runt-associated tran-scription factor 2(Runx2),osteocalcin(OCN),osteopontin(OPN),bone morphogenetic protein-2(BMP-2),Wnt-3a,β-catenin and glycogen synthase kinase-3 β(GSK-3 β)were detected by qRT-PCR.The protein expression levels of Wnt-3a,β-catenin and GSK-3βwere detected by Western blot.Results There was no significant difference in the relative activity of hPDLSCs after treated with different concentrations of 7,8-dihydroxyflavone(F=1.693,P=0.152).Compared with control group,the relative ALP activity of hPDLSCs in low-dose group,medium-dose group and high-dose group increased,the formation of calcified nodule increased,the relative mRNA levels of Runx2,OCN,OPN and BMP-2 increased,and the relative expression levels of Wnt-3a and β-catenin increased,while the relative expression level of GSK-3β decreased(P＜0.05).Compared with low-dose group,the relative ALP activity of hPDLSCs in medium-dose group and high-dose group increased,the formation of calcified nodule increased,the relative mRNA levels of Runx2,OCN,OPN and BMP-2 increased,the relative expression levels of Wnt-3a and β-catenin increased,while the relative expression level of GSK-3β decreased(P＜0.05).Compared with high-dose group,the relative ALP activity of hPDLSCs decreased in XAV939 group,the formation of calcified nodule decreased,the relative mRNA expression levels of Runx2,OCN,OPN and BMP-2 decreased,the relative expression levels of Wnt-3a and β-catenin decreased,while the relative expression level of GSK-3β increased(P＜0.05).Conclusion 7,8-dihydroxyflavone can promote the osteogenic differentiation of hPDLSCs by activating Wnt/β-catenin signaling pathway.关键词
7,8-二羟基黄酮/人牙周膜干细胞/成骨分化/成骨相关基因/Wnt/β-catenin信号通路Key words
7,8-dihydroxyflavone/human periodontal ligament stem cells/osteogenic differentiation/osteogenic related genes/Wnt/β-catenin signaling pathway分类
医药卫生引用本文复制引用
王家亮,贺莹,杨昊,贺专,董瑜,杨一帆..7,8-二羟基黄酮通过Wnt/β-catenin信号通路调控人牙周膜干细胞成骨分化[J].山西医科大学学报,2023,54(12):1638-1646,9.基金项目
陕西省重点研发计划项目(2018SF-111) （2018SF-111）
陕西省卫生健康科研基金项目(2022D050) （2022D050）
西安市科技计划项目(20YXYJ00010(4)) （20YXYJ00010(4）
