摘要
目的:探讨松果菊苷(ECH)调节CXC趋化因子配体12(CXCL12)/CXC趋化因子受体4(CXCR4)信号通路对牙髓干细胞(DPSCs)增殖、迁移和分化的影响.方法:以DPSCs为研究对象,分别经不同浓度(0.01 mg·mL-1、0.1 mg·mL-1、1 mg·mL-1)的 ECH 处理,记为 ECH 低浓度(ECH-L)组、ECH 中浓度(ECH-M)组、ECH 高浓度(ECH-H)组;以 1 mg·mL-1ECH、100 nmol·L-1 AMD3100 同时处理 DPSCs,记为ECH-H+AMD3100组,并以未经处理的DPSCs为空白对照组;MTT法检测细胞增殖;Transwell实验检测细胞迁移;Western blot检测CXCL12、CXCR4蛋白表达;成骨诱导21 d后,茜素红S染色评估矿化结节形成;碱性磷酸酶(ALP)试剂盒分析ALP活性;qRT-PCR检测牙本质基质蛋白1(DMP1)、骨钙素(OCN)、ALP mRNA表达水平.结果:与空白对照组相比,ECH-L组、ECH-M组、ECH-H组CXCL12、CXCR4蛋白表达、迁移数、ALP含量、矿化结节含量、A450值、DMP1、OCN、ALP mRNA表达水平显著增加,不同浓度ECH间均具有统计学差异(P＜0.05);与ECH-H组相比,ECH-H+AMD3100组CXCL12、CXCR4蛋白表达、迁移数、ALP含量、矿化结节含量、4450值、DMP1、OCN、ALP mRNA表达水平显著降低(P＜0.05).结论:ECH通过上调CXCL12/CXCR4信号通路促进DPSCs增殖、迁移和分化.
Abstract
Objective:To investigate the impacts of Echinacetin(ECH)on the proliferation,migration,and differentiation of dental pulp stem cells(DPSCs)by regulating the CXC chemokine ligand 12(CXCL12)/CXCR4 signaling pathway.Methods:DPSCs were studied and subjected to different concentrations of ECH(0.01 mg·mL-1,0.1 mg·mL-1,1 mg·mL-1),and they were grouped into low ECH concentration(ECH-L)group,medium ECH concentration(ECH-M)group,and high ECH concentration(ECH-H)group;DPSCs were treated with 1 mg·mL-1ECH and 100 nmol·L-1 AMD3100 simultaneously,and recorded as the ECH-H+AMD3100 group,and untreated DPSCs were used as blank control group;MTT method was applied to detect cell proliferation;Transwell experiment was applied to detect cell migration;Western blot was applied to detect the expression of CXCL12 and CXCR4 proteins;21 days after osteogenic induction,alizarin red S staining was applied to evaluate the formation of mineralized nodules;alkaline phosphatase(ALP)assay kit was applied to analyze ALP activity;qRT-PCR was applied to detect the expression levels of dentin matrix protein 1(DMP1),osteocalcin(OCN),and ALP mRNA.Results:Compared with the blank control group,the expression of CXCL12,CXCR4 protein,migration number,ALP content,mineralized nodule content,A450 value,expression levels of DMP1,OCN,and ALP mRNA in the ECH-L group,ECH-M group,and ECH-H group obviously increased,there were statistical differences among different concentrations of ECH(P＜0.05);compared with the ECH-H group,the expression of CXCL12,CXCR4 protein,migration number,ALP content,mineralized nodule content,A450value,expression levels of DMP1,OCN,and ALP mRNA in the ECH-H+AMD3100 group obviously decreased(P＜0.05).Conclusion:ECH promotes the proliferation,migration,and differentiation of DPSCs by up-regulating the CXCL12/CXCR4 signaling pathway.
维普
万方数据