目的探讨中药黄芩提取物黄芩素对人牙周膜细胞(hPDLCs)增殖和成骨基因RUNX2、BMP2表达的影响。方法原代培养hPDLCs,hPDLCs中分别加入浓度为1.25、2.50、5.00、10.00μmol/L的黄芩素作为实验组,空白组不加黄芩素。MTT法检测黄芩素对hPDLCs增殖的影响;试剂盒检测ALP的活性和表达;qRT-PCR研究黄芩素对hPDLCs RUNX2、BMP2表达的影响。结果较空白组,1.25-10.00μmol/L黄芩素作用于hPDLCs,其增殖活力略下降,但差异无统计学意义(P〉0.05);黄芩素组ALP活性和表达上升(P〈0.05),呈浓度依赖性;成骨相关基因RUNX2、BMP2 mRNA表达水平随时间延长,表达量持续上升,11 d上升最为显著(P〈0.05),且10.00μmol/L黄芩素组显著高于其余浓度组(P〈0.05)。结论黄芩素对hPDLCs骨向分化有促进作用,有可能作为牙周再生的选择之一。
Objective To evaluate the effects of baicalein extracted from Scutellaria baicalensis Georgi on the proliferation and expression of RUNX2 and BMP2 in human periodontal ligament cells( hPDLCs). Methods Baicalein at a concentration of 1.25,2.50,5.00 and 10.00 μmol / L was separately added into hPDLCs primary cultured in vitro accordingly as experimental groups. The effect of the proliferation of hPDLCs was assessed through MTT assay,while the activity and expression of Alkaline phosphatase( ALP) were assessed by kit. qRT-PCR was used to observe the mRNA expression of RUNX2 and BMP2 in hPDLCs respectively. Results Compared to control group,baicalein at different concentrations have no significant influence on the proliferation of hPDLCs. Baicalein was shown to enhance the ALP expression of hPDLCs. The mRNA level of RUNX2、BMP2 increased by baicalein in different concentrations,which reached maximum at 11 days,appear in a dose-dependent manner from 1.25 to 10 μmol / L using real-time PCR analysis.Conclusion Baicalein could promote osteogenic differentiation of hPDLCs,which may be used as one of the choices of periodontal regeneration in the future.