摘要
目的 观察补骨脂素对人牙周膜干细胞(hPDLSCs)增殖、成骨分化的促进作用,并探讨其机制.方法 将第三代hPDLSCs分为6组,5、10、25、50、100 μmol/L补骨脂素组加入5、10、25、50、100 μmol/L补骨脂素,对照组正常培养,采用CCK-8法检测细胞增殖能力,比色法检测碱性磷酸酶(ALP)活性,茜素红染色检测成骨诱导矿化结节.另取第三代hPDLSCs并分为两组,25 μmol/L补骨脂素组加入25 μmol/L补骨脂素,对照组正常培养,采用RT-PCR法检测转录因子2(Runx2)、骨桥蛋白(OPN)mRNA.结果 与对照组比较,5、10、25、50、100 μmol/L补骨脂素组细胞增殖能力和ALP活性增加,25 μmol/L补骨脂素组细胞成骨矿化结节增加(P均<0.05);与5 μmol/L补骨脂素组比较,25、50、100 μmol/L补骨脂素组细胞增殖能力及10、25 μmol/L补骨脂素组细胞ALP活性和25 μmol/L补骨脂素组细胞成骨矿化结节增加(P均<0.05);与10 μmol/L补骨脂素组比较,25、50、100 μmol/L补骨脂素组细胞增殖能力增加(P均<0.05);与25 μmol/L补骨脂素组比较,50、100 μmol/L补骨脂素组细胞ALP活性降低(P均<0.05).与对照组比较,25 μmol/L补骨脂素组Runx2、OPN mRNA相对表达量增加(P均<0.05).结论 5～100 μmol/L补骨脂素均能促进hPDLSCs的增殖和成骨分化,在5～25 μmol/L呈剂量依赖性增强,在50～100 μmol/L变化不明显;补骨脂素促进hPDLSCs的增殖和成骨分化的机制可能与调节Runx2信号通路有关.
Abstract
Objective To observe the promotional effects of psoralen on proliferation and osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs),and to explore their mechanism.Methods The third-generation hPDLSCs were divided into six groups:the control group and 5,10,25,50 and 100 μmol/L psoralen groups,respective-ly.HPDLSCs in the 5,10,25,50 and 100 μmol/L psoralen groups were treated with 5,10,25,50 and 100 μmol/L psoralen,and hPDLSCs in the control group were cultured normally.Cell proliferation capacity was measured by CCK-8 assay,alkaline phosphatase(ALP)activity by colorimetric method,and mineralization nodules by alizarin red staining.Another third-generation hPDLSCs were divided into two groups:the control group and 25 μmol/L psoralen group.Cells in the 25 μmol/L psoralen group were treated with 25 μmol/L psoralen,while cells in the control group were cultured nor-mally.Transcription factor 2(Runx 2)and osteopontin(OPN)mRNA were detected by RT-PCR.Results Compared with the control group,cell proliferation capacity and ALP activity increased in the 5,10,25,50 and 100 μmol/L pso-ralen groups,and mineralization nodules increased in the 25 μmol/L psoralen group(all P<0.05).Compared with the 5 μmol/L psoralen group,cell proliferative capacity increased in the 25,50 and 100 μmol/L psoralen groups,ALP activity increased in the 10 and 25 μmol/L psoralen groups,and mineralization nodules increased in the 25 μmol/L psoralen group(all P<0.05).Compared with the 10 μmol/L psoralen group,cell proliferation capacity increased in the 25,50 and 100 μmol/L psoralen groups(all P<0.05).Compared with the 25 μmol/L group,ALP activity decreased in the 50 and 100 μmol/L psoralen groups(all P<0.05).Compared with the control group,relative expression levels of Runx 2 and OPN mRNA increased in the 25 μmol/L psoralen group(all P<0.05).Conclusions Psoralen of 5-100 μmol/L pro-motes the proliferation and osteogenic differentiation of hPDLSCs.It shows a dose-dependent enhancement at 5 to 25 μmol/L,and there is no obvious change at 50 to 100 μmol/L.The mechanism of psoralen promoting the proliferation and osteogenic differentiation of hPDLSCs may be related to the regulation of Runx 2 signaling pathway.
基金项目
山东省自然科学基金青年基金(ZR2023QH421)
国家科技期刊平台
NSTL
万方数据