黄芩素对人牙周膜细胞骨向分化能力的影响

The effects of baicalein on promoting the osteogenesis of human periodontal ligament cells

目的:探讨中药黄芩提取物黄芩素对人牙周膜细胞(h PDLCs)增殖和成骨分化功能的影响。方法:原代培养h PDLCs,向h PDLCs中分别加入浓度为0.3125、0.625、1.25μmol/L的黄芩素作为实验组,对照组不加黄芩素。MTT法检测黄芩素对h PDLCs增殖的影响后,分别采用碱性磷酸酶(ALP)活性测定、茜素红S染色、Real-time PCR和Western Blot法研究黄芩素对h PDLCs ALP活性、矿化结节形成、I型胶原(Col-I)m RNA和蛋白表达的影响。结果:与对照组相比,各浓度黄芩素对h PDLCs增殖均无明显影响;各浓度黄芩素组均表现为ALP活性上升;同时骨向诱导21 d后可见黄芩素组矿化结节形成增加,定量分析结果显示1.25μmol/L黄芩素组形成矿化结节数量最多;Real-time PCR和Western Blot结果显示各浓度黄芩素组Col-I m RNA和蛋白表达均有上升,7 d时随浓度的增加其表达上升,1.25μmol/L黄芩素组达到最高水平。结论:黄芩素能促进h PDLCs的骨向分化能力,具有进一步研究其辅助牙周再生治疗的价值。

Objective： To evaluate the effects of baicalein extracted from Scutellaria baicalensis Georgion the function of proliferation and osteogenesis of human periodontal ligament cells （hPDLCs）. Methods： Baicalein at a concentration of 0.3125, 0.625, 1.25 μmol/L was added into hPDLCs primary cultured in vitro ac- cordingly as experimental group, the one with no baicalein as blank control group. The effect of proliferation of hPDLCs was assessed through MTT assay. Alkaline phosphatase （ALP） activity assay, Alizarin red S staining, RT-qPCR and Western Blot were used to observe the ALP activity, formation of mineral nodules, mRNA and protein expression of collagenase-I （Col-I） of hPDLCs, respectively. Results： Compared to the control group, baicalein at different concentrations had no significant influence on the proliferation of hPDLCs. Baicalein was shown to enhance the ALP activity of hPDLCs. After 21 days of osteogenic induction, more mineral nodules could be observed compared to the control group, which increased in a dose-dependent manner at the concentra- tion of 0.3125-1.25 μmol/L baicalein according to the results of quantification. The mRNA and protein level of Col-I increased by baicalein in different concentration, which reached maximum at 7 days, appearring in a dose- dependent manner from 0.3125 to 1.25 ktmol/L using Western Blot and Real-time PCR analysis. Conclusion： Baicalein can promote hPDLCs to differentiate into osteoblasts, which is of great value for further study acting as the adjuvant drug for oeriodontal regenerative therapy.