目的:通过观察中药黄芩的有效成份黄芩苷对人牙周膜细胞(human periodontal ligament cells,hPDLCs)增殖、矿化成骨等功能活性的影响,研究黄芩对人牙周膜细胞的调节作用。方法:采用组织贴块联合胶原酶消化的方法,体外分离、培养、纯化和鉴定hPDLCs后,通过四甲基偶氮唑蓝(MTT)比色法、酶联免疫吸附、半定量逆转录聚合酶链反应(RT-PCR)等方法,检测不同浓度的黄芩苷(10、1.0、0.1、0.01mg/L)对hPDLCs增殖、矿化成骨、骨保护素(osteoprotegerin,OPG)、细胞核因子-κB受体活化因子配基(receptor activator of nuclear factor-κB ligand,RANKL)mRNA表达等方面的影响。结果:0.1mg/L的黄芩苷对于增加人牙周膜细胞的增殖活性、碱性磷酸酶活性和Ⅰ型胶原蛋白的表达作用最强;适宜浓度的黄芩苷可以降低RANKL/OPGmRNA比值。结论:黄芩苷能促进hPDLCs的增殖和成骨作用,该作用可能与RANKL/OPG信号通路相关。
Objective:To observe the effect of baicalin on human periodontal ligament cells'(hPDLCs) function and activity of proliferation,bone formation and so on.Methods:The hPDLCs' abruption,culture,purification and identification was by tissue culture method in the combination of enzyme digestion.To detect the effect of baicalin in different concentrations(10,1.0,0.1,0.01 mg/L) on hPDLCs' proliferation,bone formation,osteoprotegerin(OPG),receptor activator of nuclear factor-κB ligand(RANKL) mRNA expression,we used the methods including MTT method,enzyme linked immunosorbent assay and RT-PCR.Results:The baicalin concentrations of 0.1 mg/L could enhance the proliferation of hPDLCs,activity of alkaline phosphatase(ALP) and expression of collagen protein-Ⅰ,depress the ratio of RANKL/OPG mRNA.Conclusions:Baicalin had the capability of promoting hPDLCs' proliferation and bone formation,which reason may be the reduction of RANKL/OPG mRNA.