基质细胞趋化因子-1对人牙周膜干细胞趋化因子受体——CXC亚家族受体4表达的影响研究

目的:证实人牙周膜干细胞(human periodontal ligament stem cells,hPDLSCs)表达趋化因子受体——CXC亚家族受体4(cysteine X cysteine receptor 4,CXCR4)及探究趋化因子基质细胞趋化因子-1(stromalcell-derived factor1,SDF-1)和阻断剂AMD3100(商品名:普乐沙福)对CXCR4表达的影响,从而为探究牙周膜干细胞生物学效应的影响提供理论基础。方法:采用消化组织块法联合有限稀释法分离纯化得到hPDLSCs,取第3代hPDLSCs随机分为3组:阴性对照组、SDF-1组(200μg/L SDF-1),SDF-1+AMD3100组(200μg/L SDF-1+10 mg/L AMD3100)。利用Western blot检测CXCR4在hPDLSCs上的表达及在SDF-1、AMD3100作用下CXCR4表达的变化。结果:1.筛选后的hPDLSCs可被诱导分化为成骨细胞和成脂细胞,证实其具有多向分化潜能;利用流式细胞仪检测其细胞表型鉴定其符合牙周膜干细胞的免疫表型。2.Western blot检测结果显示hPDLSCs上表达CXCR4,且应用SDF-1后上调CXCR4的表达,SDF-1+AMD3100组无明显变化。结论:1.hPDLSCs可从新鲜离体牙的牙周膜中培养获得,经有限稀释法纯化后鉴定为间充质来源。2.hPDLSCs上表达CXCR4,SDF-1通过上调hPDLSCs上CXCR4的表达,AMD3100可阻断SDF-1与其受体CXCR4结合。

Effect of SDF-1 on CXCR4 Expression of Human Periodontal Stem Cell Chemokine Receptor

Objective:To confirm the effect of human periodontal ligament stem cells(hPDLSCs)on expression of chemokine receptor CXCR4,and the effect of chemokine SDF-1 and blocker AMD3100 on expression of CXCR4.Methods:hPDLSCs were isolated and purified by digestible tissue block method combined with limited dilution method.The third generation of hPDLSCs were randomly divided into three groups:negative control group,SDF-1 group(200μg/L SDF-1),and SDF-1+AMD3100 group(200μg/L SDF-1+10 mg/L AMD3100).Western blotting was used to detect the expression of CXCR4 on hPDLSCs and the expression of CXCR4 under the action of SDF-1 and AMD3100.Results:The selected hPDLSCs could be induced to differentiate into osteoblasts and adipocytes.The cell phenotype was detected by flow cytometry to identify the immunophenotype consistent with periodontal ligament stem cells.Western blotting analysis showed that CXCR4 was expressed on hPDLSCs,and the expression of CXCR4 was up-regulated after SDF-1.There was no significant change in SDF-1+AMD3100 group.Conclusion:hPDLSCs could be obtained from the periodontal ligament of fresh isolated teeth and purified as a mesenchymal source after purification by limiting dilution method.The CXCR4 receptor was expressed on hPDLSCs.SDF-1 up-regulated CXCR4 receptor on hPDLSCs,and AMD3100 blocked the binding of SDF-1 to CXCR4 receptor.