中药黄芩苷对牙髓干细胞增殖及成牙成骨分化能力的影响

摘要

Objective:To investigate the effect of baicalin on the proliferation and odento/osteogenic differentiation of huaman dental pulp stem cells ( hDPSCs) in vitro.Methods:hDPSCs were isolated from dental pulp and then were cultured in alpha minimun essen-tial medium(α-MEM) supplimented with or without baicalin.MTT colorimetric method was used to examine the proliferation ability of hDPSCs.The effect of baicalin on the odento/osteogenic differentiation was observed by alkaline phosphotase( ALP) activity and West-ern blot.Results:MTT assay showed that there was no significant difference between baicalin-treated and untreated control cells ( P>0.05), and ALP activity assay demonstrated that 0~20μmol/L baicalin enhanced the ALP activity of DPSCs(P<0.01), and West-ern blot revealed that the odento/osteogenic marker (RUNX2, DSP, OSX, OPN, OCN) was significantly upregulated in baicalin-trea-ted cells compared with the untreated cells.Conclusions:Baicalin has no effect on the proliferation of DPSCs, but enhances the ca-pacity of odento/osteogenic differentiation.%目的：探讨中药黄芩苷对牙髓干细胞的增殖、成牙／成骨分化能力的影响。方法：酶消化法原代培养人牙髓干细胞，采用甲基噻唑基四唑（ MTT）比色法检测黄芩苷对人牙髓干细胞增殖的影响，碱性磷酸酶活性检测及Western blot等方法检测黄芩苷作用于人牙髓干细胞后，其成牙／成骨分化指标，包括核心结合因子（runt-related transcription factor 2，RUNX2）、牙本质涎蛋白（ dentin sialoprotein，DSP）、Osterix（ OSX）、骨桥蛋白（ osteopontin，OPN）、骨钙素（ osteocalcin，OCN）的变化。结果：MTT检测结果显示，0～20μmol／L黄芩苷作用于牙髓干细胞后其增殖能力与对照组相比无明显差异（P＞0．05）；而碱性磷酸酶活性检测显示：0～20μmol／L黄芩苷各浓度组细胞ALP活性比对照组明显增加（P＜0．01）；Western blot结果表明：与对照组相比，20μmol／L黄芩苷可增强牙髓干细胞RUNX2、DSP、OSX、OPN、OCN等成牙／成骨相关蛋白的表达。结论：黄芩苷对牙髓干细胞的增殖无明显影响，但可促进其成牙／成骨分化能力。